CN104257756A - Application of cortex mori fatty oil in preparation of hypoglycemic agent - Google Patents

Abstract

The invention relates to the application of Morus alba fat oil in the preparation of hypoglycemic drugs, which can effectively solve the application problem of Morus alba fat oil in the preparation of hypoglycemic drugs. For the application in sugar medicine, the Morus alba fatty oil is immersing the Morus alba in water, decocting for 30-40min, removing the dregs to obtain the Morus alba water extract, concentrating and drying the Morus alba water extract, To obtain a dried product, add 10 times the amount of petroleum ether to the dried product and reflux for extraction for 6 hours, filter, recover the filtrate, evaporate the petroleum ether to obtain the petroleum ether extraction part, evaporate the water at 100 ° C, and obtain Morus Alba Fatty Oil, the present invention The preparation method of Morus alba fatty oil is simple, and the same or similar results have been obtained after many tests. In the past, Morus alba fatty oil has no obvious medicinal value, and it is generally discarded as a non-medicinal part. The invention enables waste to be utilized, expands medicinal resources, and opens up a new medicinal use of Morus alba fat oil.

Description

Application of a kind of Morus alba fat oil in the preparation of hypoglycemic drugs

technical field

The invention relates to the field of medicine, in particular to the application of Morus alba fat oil in the preparation of hypoglycemic drugs. the

Background technique

Diabetes is a chronic endocrine disease with extremely high morbidity and mortality, and its onset is often accompanied by hypertension, hyperlipidemia and hyperuric acid. At present, the most important thing in the treatment of diabetes is to effectively and stably control blood sugar, which may require life-long medication. However, long-term administration of oral hypoglycemic chemical drugs will inevitably produce many side effects and cause great damage to the liver. Therefore, the development of high-efficiency, low-toxic hypoglycemic drugs from traditional Chinese medicine has good development prospects and great significance. It has been reported that there are dozens of active ingredients with hypoglycemic effect, mainly polysaccharides, alkaloids, triterpenoids and flavonoids. the

Morus alba L. is the dry root bark of the Moraceae plant Morusalba L., which was first recorded in "Shen Nong's Materia Medica", listed as a middle grade, and has been recorded in successive dynasties of herbal medicines. Cortex Mori is sweet and cold in nature, and belongs to the lung meridian. It has the effects of purging the lung, relieving asthma, diuresis and reducing swelling. Modern pharmacological research has found that Cortex Morus alba mainly has the effects of lowering blood sugar, diuresis, antitussive, expectorant, asthma, anti-inflammatory and analgesic, immune regulation, anti-tumor, anti-bacterial, anti-viral, lowering blood pressure and affecting the digestive system. the

There are records about Morus alba treating diabetes in ancient Chinese medicine classics and prescriptions, and its hypoglycemic effect on diabetic animal models has also been reported. Wang Ning and others explored the mechanism of Morus Alba Cortex and found that one of the mechanisms of Morus Alba Cortex preventing and treating type 2 diabetes may be achieved by promoting glucose metabolism in peripheral tissues and improving the sensitivity of liver cells to insulin. During Zhang Qian's screening of the inhibitory effect of water and alcohol extracts of 20 traditional Chinese medicines on α-glucosidase, it was found that the extract of Morus alba extract had the strongest inhibitory effect, which was similar to acarbose, that is, it could be non-competitive. Inhibit the activity of α-glucosidase, thereby reducing the peak blood sugar in normal mice and diabetic mice after maltose loading, regulating blood sugar levels, improving symptoms such as polydipsia, polyphagia, and weight loss in diabetic mice, and slowing down intestinal maltose, etc. Digestion and absorption of disaccharides. The alkaloids in Cortex Morus alba can inhibit α-glucosidase, and can effectively reduce fasting blood sugar in diabetic mice, especially when combined with green tea polyphenols, it has a certain superimposed effect. Zhong Guolian and others studied the hypoglycemic effect of the extract obtained after the Morus alba water decoction was precipitated with 95% ethanol, and found that the Morus alba water-alcohol extract had a hypoglycemic effect on experimental diabetic rats, but had no effect on blood lipids. Influence. the

The chemical composition of Morus alba is complex, mainly containing flavonoids, stilbenes, coumarins, lignans, alkaloids and fatty oils, etc. Researches mainly focus on flavonoids and alkaloids, but there are few studies on the pharmacological effects of other components of Morus alba. Fatty oil is the esterification product of fatty acid and glycerol, which is ubiquitous in nature. At the beginning of the 1980s, the research on the chemical constituents and pharmacological activity of fatty oil of Chinese herbal medicine attracted the common attention of scholars, but the research on fatty oil of Morus alba has not been reported yet. the

Contents of the invention

In view of the above situation, in order to solve the defects of the prior art, the purpose of the present invention is to provide a kind of Morus alba fat oil in the preparation of hypoglycemic drugs, which can effectively solve the application of Morus alba fat oil in the preparation of hypoglycemic drugs. question. the

The technical scheme solved by the present invention is the application of Morus alba fatty oil in the preparation of hypoglycemic drugs. The Morus alba fatty oil is steam distilled to obtain the volatile oil component, its water extract is concentrated, and the Extract with 2 times the amount of petroleum ether, concentrate the petroleum ether layer, recover the solvent, and dry in vacuum for 6 hours to obtain Morus alba fatty oil. the

The preparation method of Morus Alba Fatty Oil of the present invention is simple, and through repeated tests, all obtained the same or similar results. In the past, Morus Alba Fatty Oil had no obvious medicinal value, and was generally discarded as a non-medicinal part. , the present invention makes use of waste, expands medicinal resources, and opens up a new medicinal use of Morus alba fat oil. In addition, the Morus alba fat oil has less dosage, is convenient to take, and has great clinical significance. the

Description of drawings

Fig. 1 is the technological flow chart of each dismantling part of Cortex Mori of the present invention. the

Fig. 2 is a graph of cluster analysis of samples of Cortex Morus alba 30%, 50%, and 80% ethanol elution parts of the present invention. the

Fig. 3 is the principal component analysis diagram of samples of Morus alba 30%, 50%, and 80% ethanol elution parts of the present invention. the

Fig. 4 is an analysis chart of orthogonal partial least squares analysis of samples of Morus alba 30%, 50%, and 80% ethanol elution parts of the present invention. the

Fig. 5 is a schematic diagram showing the effect of Morus alba fatty oil of the present invention on the body weight of diabetic mice. the

Fig. 6 is a schematic diagram showing the effect of Morus alba fatty oil of the present invention on the diet of diabetic mice. the

Fig. 7 is a schematic diagram showing the effect of Morus alba fatty oil of the present invention on the water intake of diabetic mice. the

Fig. 8 is a schematic diagram showing the effect of Morus alba fatty oil of the present invention on FBG in diabetic mice. the

Fig. 9 is a schematic diagram showing the effect of Morus alba fatty oil of the present invention on insulin in diabetic mice. the

Fig. 10 is a schematic diagram showing the effect of Morus alba fatty oil of the present invention on C-peptide in diabetic mice. the

Fig. 11 is a schematic diagram showing the effect of Morus alba fatty oil of the present invention on TC of diabetic mice. the

Fig. 12 is a schematic diagram showing the effect of Morus alba fatty oil of the present invention on TG in diabetic mice. the

Fig. 13 is a schematic diagram of the effect of Morus alba fatty oil of the present invention on the pathological changes of liver histopathology in diabetic mice (HE staining, ×400). the

Fig. 14 is a schematic diagram of the effect of Morus alba fatty oil of the present invention on the histopathological changes of pancreas in diabetic mice (HE staining, ×400). the

Fig. 15 is a total ion chromatogram of fatty oil components of Cortex Mori of the present invention. the

Detailed ways

The specific implementation manners of the present invention will be described in further detail below in conjunction with the accompanying drawings and examples. the

Example 1

The application of Morus alba fat oil in the preparation of hypoglycemic drugs, the Morus alba fatty oil is immersing 100g of Morus alba in water, decocting for 30min, removing the dregs to obtain Morus alba water extract, and Morus alba Concentrate the water extract, add 2 times the amount of petroleum ether for extraction, concentrate the petroleum ether layer, recover the solvent, and dry in vacuum for 6 hours to obtain Morus alba fatty oil. the

Example 2

The application of Morus Alba Fatty Oil in the preparation of hypoglycemic drugs, the Morus Alba Fatty Oil is immersing Morus Alba 500g in water, decocting for 35min, removing the medicine residues to obtain the Morus Alba water extract, and Morus Alba Concentrate the water extract, add 2 times the amount of petroleum ether for extraction, concentrate the petroleum ether layer, recover the solvent, and dry in vacuum for 6 hours to obtain Morus alba fatty oil. the

Example 3

The application of Morus alba fatty oil in the preparation of hypoglycemic drugs, the Morus alba fatty oil is immersing 1000g of Morus alba in water, decocting for 40min, removing the dregs to obtain Morus alba water extract, and Morus alba Concentrate the water extract, add 2 times the amount of petroleum ether for extraction, concentrate the petroleum ether layer, recover the solvent, and dry in vacuum for 6 hours to obtain Morus alba fatty oil. the

According to the traditional method of using traditional Chinese medicine, the present invention adopts water decoction combined with extraction, alcohol precipitation, macroporous adsorption resin and other technologies to divide Morus alba water decoction into the following 6 parts: volatile oil part, petroleum ether extraction part (i.e. this Invention fatty oil part), alcohol precipitation part, 30% ethanol elution part, 50% ethanol elution part, 80% ethanol elution part, and after confirming that the components of each part do not cross each other through various detection methods, each part is tested Through the screening of hypoglycemic activity, it was found for the first time that the fat oil fraction of Morus alba can significantly reduce the fasting blood sugar of diabetic mice, improve the symptoms of "three more and one less" in diabetic mice, and can improve the lipid metabolism of diabetic mice to a certain extent disorders and peripheral tissue damage. This also provides a reference for the development and utilization of the traditional Chinese medicine Morus alba fatty oil. the

The invention provides a preparation method for each split part of Cortex Morus alba, through the research on the hypoglycemic effect of the mouse model of diabetes, it is clear that the fatty oil of Cortex Morus alba is the effective part to exert the effect of lowering blood sugar, and the chemical components therein are analyzed, It lays the foundation for expanding the medicinal parts of Cortex Mori and ascertaining the material basis of its function. The preparation method of Morus Alba Fatty Oil of the present invention is simple, through repeated tests, all obtained the same or similar results, in the past Morus Alba Fatty Oil did not find obvious medicinal value, generally as non-medicinal parts often used Abandoned, the present invention enables waste to be utilized, expands medicinal resources, and opens up a new medicinal use of Morus alba fat oil. The relevant test data are as follows:

1 Experimental materials

1.1 Animals:

90 male Kunming mice, weighing 18-22 g, were provided by Shandong Lukang Experimental Animal Center. They were raised in a constant temperature (18-22° C.), relative humidity of 65%-70%, photoperiod of 12h:12h, free-to-eating and drinking environment. Adaptive feeding was used for experiments after 1 week. the

1.2 Drug preparation:

Morus alba L. was purchased from Zhengzhou Chinese Medicinal Materials Market, and was identified as the dry root bark of Moraceae Morus alba L. by Professor Dong Chengming, Department of Pharmacognosy, School of Pharmacy, Henan University of Traditional Chinese Medicine. the

Extraction method of each part of Morus alba: self-made, see 2.1 for the preparation method. the

1.3 Reagents:

Streptozotocin (STZ) (lot number: S0130, Sigma, USA); metformin (lot number: 1302071, Sino-US Shanghai Bristol-Myers Squibb Pharmaceutical Co., Ltd.); Company); Insulin ELISA Kit (Lot No.: 20140401A, R&D Company); C-peptide ELISA Kit (Lot No.: 20140401A, R&D Company); Total Cholesterol (TC) Detection Kit (Lot No. 20130116, Beijing Beihua Kangtai Clinic Reagent Co., Ltd.); triglyceride (TG) detection kit (batch number 20130115, Beijing Beihua Kangtai Clinical Reagent Co., Ltd.); sodium citrate (Tianjin Dengke Chemical Reagent Co., Ltd.); citric acid (Tianjin Hengxing Chemical Reagent Co., Ltd. Manufacturing Co., Ltd.); other various chemical reagents were commercially available of analytical grade. the

1.4 Instruments:

ChemPattern software (Kermain Beijing Technology Co., Ltd.); 680 type microplate reader (BIO-RAD); Arium611VF super combination ultrapure water device (SARTORIUS); KDC-160HR high-speed low-temperature refrigerated centrifuge (KUST Innovation Co., Ltd.) ; SHHW21.420-C type three water tank (Beijing Changyuan Experimental Equipment Factory); Guohua SK-1 fast mixer (Changzhou Guohua Electric Co., Ltd.); AB204-N type 1/10,000 electronic balance; DZKW- 4. Electronic constant temperature water bath (Beijing Zhongxing Weiye Instrument Co., Ltd.); Agilent7890/7000B triple quadrupole gas spectrometer (Agilent, USA); standard glass Soxhlet extractor (Shanghai Yuming Instrument Co., Ltd.); Q-250A3 Pulverizer (Ruida International); SB-1000 rotary evaporator (Shanghai Ailang Instrument Co., Ltd.). the

2 experimental plan

2.1 Preparation method of each split part of Cortex Morus alba:

Cortex Mori steam distillation obtains volatile oil component, its water extract is concentrated, adds 2 times amount of petroleum ether to extract, petroleum ether layer is concentrated, after reclaiming solvent, vacuum drying 6h obtains petroleum ether component (that is Cortex Mori of the present invention Fatty oil), the oil yield was 2.99%. Then put DiaionHP-20 macroporous adsorption resin on the water extract layer to obtain 30% ethanol elution fraction, 50% ethanol elution fraction, 80% ethanol elution fraction and alcohol precipitation fraction (polysaccharide fraction), The process flow is shown in Figure 1. the

2.2 Study on the non-crossover of components contained in each split part of Morus alba

According to the preparation method under 2.1, 10 batches of split parts were prepared respectively, and the 10 batches of split parts were detected by HPLC-PDA, HPLC-ELSD and UPLC-DAD methods, and the advantages of different types of detectors were complementary , from different angles and levels to examine the crossover of the chemical components of each component. At the same time, the reproducibility of the separation process was investigated through the scale-up experiment of the separation process, which provided a guarantee for the non-crossover of each separation component in the large-scale preparation, so as to ensure that the drug preparation method used in the subsequent pharmacological experiments was stable and the results were true. reliable. the

2.3 Model establishment

The mice were fed adaptively for 1 week, and 10 mice were randomly selected as the normal control group according to the principle of body weight balance, and the remaining 80 mice were intraperitoneally injected with 170 mg/kg STZ (prepared with 0.1 mol/L citric acid-sodium citrate buffer solution with pH 4.21 into 1% solution, just before use

Matching, ice bath operation), the normal control group was intraperitoneally injected with the same dose of citric acid-sodium citrate buffer. After injecting STZ for 72 hours, fast for 4 hours, take blood from tail vein, centrifuge at 3500r/min for 10min, separate serum, and detect blood glucose by glucose oxidase method. The modeling standard is that the fasting blood glucose is greater than or equal to 11.1mmo1/L. Model mice were selected for experiments. the

2.4 Experimental grouping and administration method

Rats with successful modeling were grouped according to the principle of blood sugar balance. The normal control group and the model control group were given distilled water, the metformin group was given 270 mg/kg metformin, the fat oil group was given 250 mg/kg fat oil, and the normal CMC control group and model CMC control group were given the same dose of CMC solvent. Continuous administration for 4 weeks. After the administration was over and blood samples were collected, the mice were sacrificed by cervical dislocation, the liver was quickly removed, and the same lobe was cut out, immersed in formalin fixative, and the rest was quickly frozen in liquid nitrogen. Store at ℃ for later use. the

2.5 Determination of body weight, water intake, food intake, and fasting blood glucose (FBG) during administration

The changes in body weight, water intake, and food intake of rats in each group were recorded before and during the administration. After 72 hours of intraperitoneal injection of STZ and 2 and 4 weekends of treatment, fast for 4 hours, blood was collected from tail vein, and serum was separated by centrifugation at 3500r/min for 10 minutes, and FBG was detected by glucose oxidase method. the

2.6 Determination of insulin and C-peptide

After 4 weeks of administration, fast for 12 hours, blood was collected from the retro-orbital venous plexus, and the serum was separated by centrifugation at 3500r/min for 10 minutes, and the serum insulin and C-peptide levels were detected according to the kit instructions. the

2.7 Determination of blood lipids

After 4 weeks of administration, fast for 12 hours, take blood from the retro-orbital venous plexus, centrifuge at 3500r/min for 10min to separate the serum, and detect the content of serum TC and TG according to the kit instructions. the

2.8 Histopathological observation of liver and pancreas

Liver and pancreas tissues were dehydrated with graded ethanol, cleared in xylene, soaked in wax, embedded in paraffin, sectioned, routinely stained with HE, and observed for pathological changes with a light microscope. the

2.9 Statistical processing

Experimental data with Said that SPSS18.0 was used for statistical processing, and one-way analysis of variance (One-WayANOVA) was used to compare the differences between groups. P<0.05 means the difference is significant, and P<0.01 means the difference is extremely significant.

2.10 Composition analysis of Morus alba fat oil

Take an appropriate amount of Morus alba fat oil (about 100mg) in a 10mL stoppered graduated test tube, add 4ml of n-hexane, then add 1mL of 0.8ml/L sodium hydroxide methanol solution, vortex for 5min, add 0.5g of sodium bisulfate to dry, and centrifuge After 10 minutes, the supernatant was taken and passed through a 0.22 μm filter membrane for GC/MS analysis. the

Gas chromatography conditions: the chromatographic column is a DB-5 quartz capillary column (30m×0.25mm, 0.25μm), the carrier gas is high-purity helium (99.999%), the helium flow rate is 1.0mL/min; the inlet temperature is 280 ℃; split injection, split ratio 30:1, injection volume 0.1 μL; temperature rise program is 160°C, keep for 3 minutes, then raise the temperature to 240°C at 4°C/min, then rise to 245°C at 1°C/min, and finally The temperature was raised to 290°C at 4°C/min, and kept for 10 minutes, the temperature of the vaporization chamber was 280°C, and the interface temperature was 280°C. the

Mass spectrometry conditions: collision gas flow rate: helium 2.25ml/min; nitrogen 1.5ml/min; ion source: electron bombardment source, electron energy 70eV; scanning range 45-400amu, quadrupole temperature 150°C, ion source temperature 230°C, Solvent delay: 4.0min, GC/MS interface temperature 280°C. the

3 Experimental results

3.1 Research on the components contained in each part of Morus alba Cortex without crossing each other

The components contained in the parts of Cortex Morus alba were studied independently by HPLC-PDA, HPLC-ELSD and UPLC-DAD methods. Because it is impossible for the components of fatty oil part, volatile oil part, polysaccharide part (alcohol precipitation part) and water-soluble parts to cross, the parts that may cross are water elution parts, 30%, 50% and 80% ethanol elution parts. Here, taking the results of HPLC-PDA as an example (the results of HPLC-ELSD and UPLC-DAD are not shown in the figure), a complementary crossover study was carried out on the possible crossover sites. the

3.1.1 HPLC-PDA data cluster analysis

According to the separation process under item 2.1, 10 batches of 30%, 50%, and 80% ethanol elution parts of the test sample were repeatedly prepared, and the HPLC-PDA condition experiment was performed with the decoction of Cortex Mori, and the chromatogram and chromatographic data were recorded for 100 min. Import the ChemPattern software, and use the variable class averaging method for cluster analysis. The results were divided into 3 categories, each of which was 10, which proved that the elution sites of 30%, 50%, and 80% ethanol clustered into one category without intersecting each other. The result is shown in Figure 2. the

3.1.2 Principal component analysis of HPLC-PDA data

Same as 3.1.1, import the obtained chromatographic data into ChemPattern software, and use principal component analysis, the sample can be clearly divided into 3 regions, and every 10 constitutes a region, which proves that 30%, 50% and 80% ethanol elution parts are clustered into One category, not intersecting each other. The result is shown in Figure 3. the

3.1.3 Orthogonal partial least squares analysis of HPLC-PDA data

The same as 3.1.1, the analysis of orthogonal partial least squares method was carried out, the sample can be clearly divided into 3 regions, and every 10 constitutes a region, which proves that 30%, 50% and 80% ethanol elution parts are clustered into one Classes do not intersect each other. The result is shown in Figure 4. 3.2 Effects of Morus alba fat oil on body weight, drinking water, food intake and FBG of diabetic mice

During the experiment, the weight of mice in the normal group increased steadily; after injection of STZ, compared with the mice in the normal group, the body weight of the mice in the model group decreased significantly (p<0.05). The body weight of the mice in the administration group was slightly higher than that of the model group, although there was no statistical difference (P>0.05), but during the entire administration period, the body weight remained basically constant and had a tendency to increase, as shown in Table 1 and Figure 5 . the

After modeling, the diet of the mice in the model group was significantly higher than that of the normal group (p<0.01), and after two weeks of treatment, the diet of the mice in the fatty oil group was significantly lower than that of the model group (p<0.01), see Table 2, Figures 6 and 7. the

After modeling, the amount of drinking water of the mice in the model group was significantly higher than that of the normal group (p<0.01), while after treatment, the amount of drinking water of the mice in the fatty oil group decreased significantly, and compared with the model group There are extremely significant differences (p<0.01), see Table 3 and Figure 4. For the effect on FBG, after four weeks of administration and treatment, the FBG of the mice in the fatty oil group was significantly reduced, which was significantly different from that in the model group, as shown in Table 4 and Figure 8. the

Table 1 The effect of Morus alba fat oil on the body weight of diabetic mice

Note: Compared with the normal group, #=P<0.05, ##=P<0.01; compared with the model group, *=P<0.05, **=P<0.01. the

Table 2 Effect of Morus alba fat oil on diet of diabetic mice

Note: Compared with the normal group, #=P<0.05, ##=P<0.01; compared with the model group, *=P<0.05, **=P<0.01. the

Table 3 Effect of Morus Alba Fatty Oil on Drinking Water of Diabetic Mice

Note: Compared with the normal group, #=P<0.05, ##=P<0.01; compared with the model group, *=P<0.05, **=P<0.01. the

Table 4 Effect of Morus alba fat oil on FBG in diabetic mice

Note: Compared with the normal group, #=P<0.05, ##=P<0.01; compared with the model group, *=P<0.05, **=P<0.01. the

3.3 Effect of Morus Alba Fatty Oil on Glucose Regulating Hormone in Diabetic Mice

Compared with the normal group, the C-peptide level of the mice in the model group was significantly increased (p<0.05). ), but there was no significant difference in insulin levels, see Table 5, Figures 9 and 10. the

Table 5 Effect of Morus alba fat oil on glucose-regulating hormone in diabetic mice

Note: Compared with the normal group, #=P<0.05, ##=P<0.01; compared with the model group, *=P<0.05, **=P<0.01. the

3.4 Effect of Morus Alba Fatty Oil on Blood Lipid of Diabetic Mice

Compared with the normal group, the TC and TG of the mice in the model group were significantly increased (p<0.05, p<0.01). After four weeks of treatment, the TC and TG of the mice in the fatty oil group were significantly lower than those in the model group (p<0.05), see Table 6, Figures 11 and 12. the

Table 6 Effect of Morus alba fat oil on TC and TG in diabetic mice

Note: Compared with the normal group, #=P<0.05, ##=P<0.01; compared with the model group, *=P<0.05, **=P<0.01. the

3.5 Effect of Morus Alba Fatty Oil on Liver Histopathological Changes in Diabetic Mice

Under the light microscope, the hepatic lobule structure in the normal group was clear and complete, the central vein was large and the wall was thin, the liver cells were arranged into hepatic cords, distributed radially around the central vein, the cell cords were arranged neatly, the nucleus structure was clear, and the liver cells had no obvious lesions; In the model group, the liver tissue structure was disordered, irregularly arranged, mild edema, and a large number of cell necrosis; after 4 weeks of drug treatment, compared with the model group, the liver cell structure in the fat oil group had basically tended to normal , the cell morphology returned to normal, as shown in Figure 13. Observation under the light microscope, the islets of mice in the normal group were round and elliptical cell clusters with round outlines, plump islands, uniform distribution of islet cells, oval nuclei, abundant cytoplasm, and the number of islets and cells in the islets were higher than many. In the model group, the islets of the mice in the model group shrank significantly, the outline was not round, the arrangement was sparse and irregular, the nuclei shrank, the number of islets and the number of cells in the islets decreased significantly, the volume shrank, inflammatory cells appeared, and there were necrosis. After four weeks of administration, the morphology of the pancreas of the mice in the fatty oil group was slightly relieved, but the morphology of the islets was not significantly improved, as shown in Figure 14.

3.6 Analytical results of Morus alba fat oil composition

According to the analysis conditions under item "2.10", GC-MS analysis was carried out on the fatty oil components of Morus alba, and the results are shown in Figure 15. the

Perform a mass spectral library search on the mass spectra of each peak in the total ion chromatogram, select the top 10 possible substances with high similarity, and search the literature through http://webbook.nist.gov/chemistry/ (DB-5 column) Get its retention index KI worth the literature value, and preliminarily determine the chemical composition. The retention index KI literature results are compared with the retention index KI calculation results. The chemical structure with the highest similarity between the mass spectrum and the retention index KI value is the best identification result. The relative percentage content of each volatile component was obtained by the peak area normalization method, and the results are shown in Table 7.

Table 7 GC-MS analysis results of chemical components of Morus alba fatty oil

4 Conclusion

The invention establishes the type I diabetes model by one-time intraperitoneal injection of large dose of streptozotocin. Streptozotocin is a DNA alkylating agent glucosamine-nitrosourea (glucosamine-nitrosourea), which can enter cells independently through GLUT2 glucose transport protein. Highly selective toxicity to insulin-induced β-cells in pancreatic islets. Streptozotocin is toxic to GLUT2-positive neuroendocrine tumor cells, acts as a nitric oxide donor in pancreatic islets, and induces the death of insulin-producing cells. Streptozotocin is relatively less toxic to body tissues and has a high animal survival rate. It is currently a drug widely used at home and abroad to prepare animal models of diabetes. After the injection, it was observed that the humidity of the bedding of the mice increased significantly, and the amount of food and drink increased significantly. Blood glucose and blood lipid levels were found to be abnormal by taking blood from the tip of the tail, indicating that the modeling was successful. the

When the present invention screens the hypoglycemic effect of each split part of Cortex Morus alba, it is first found that the fatty oil part has a better hypoglycemic effect on the diabetic mouse model, and then the fatty oil of Cortex Morus alba is analyzed by GC-MS, from which 27 main components were separated and detected, and the sum of their relative contents accounted for 93.79% of the total detected compounds. The substance with the most abundant content was squalane, accounting for 50.40%, followed by α-amyresin (10.86%), linoleic acid Methyl ester (9.33%), β-amyloid (4.84%), methyl palmitate (3.08%), methyl oleate (1.76%), etc. Modern studies have shown that α-amyresin has anti-tumor and anti-atherosclerotic effects, methyl linoleate has anti-inflammatory and immune-regulating effects, and squalane, methyl palmitate and methyl oleate The application is limited to fabric skin care, acaricides and herbicides respectively, and there is no report of hypoglycemic effect in relevant data, which proves the feasibility of the present invention. the

The typical symptoms of diabetes are "three excesses and one less", that is, polydipsia, polyphagia, polyuria, and weight loss. The experimental results of the present invention show that the body weight of the mice in the model group drops significantly, and though the body weight of the mice in the fat oil group has no significant difference compared with the model group, it is basically stable during the administration and has an upward trend; this may be related to the It is related to the regulation of lipid breakdown, gluconeogenesis of lipids and proteins, and the improvement of the body's metabolic function. During the experiment, the amount of food and water consumed by the mice in the model group continued to increase significantly, while the amount of food and water consumed by the mice in the fatty oil group decreased significantly after administration, and there was a significant difference from the model group. By observing the mouse litter, the humidity in the model group continued to be high, the humidity in the fatty oil group was high before and at the beginning of administration, and the humidity in the middle and late stages of administration decreased significantly. It shows that fatty oil can improve the symptoms of "three excesses and one deficiency" in diabetic mice. the

Hyperglycemia is the main risk factor for diabetes. Hyperglycemia not only inhibits insulin secretion, but also inhibits insulin-stimulated glucose transport and glycogen synthesis. That is, chronic hyperglycemia can induce and aggravate insulin secretion defects and insulin resistance. Make blood sugar rise again, forming a vicious circle. This phenomenon is called "glucotoxicity". FBG reflects the level of fasting blood glucose and is the most commonly used detection index for diabetes. It reflects the function of pancreatic beta cells and generally represents the secretion function of basal insulin. Therefore, monitoring FBG is a basic indicator for judging the level of blood sugar control, and has always been the focus of clinical treatment of diabetes and understanding the progress of diabetes. The experimental results showed that after injection of STZ, the blood glucose level of the mice in the model group was significantly increased, which was significantly different from that in the normal group (P<0.01); after four weeks of administration, the fat oil could significantly reduce the fasting blood glucose level of the mice . It shows that fatty oil can regulate and improve blood sugar level. the

Insulin is the only hypoglycemic hormone in the body. It is secreted by pancreatic β cells and mainly acts on the liver, skeletal muscle and adipose tissue to regulate the metabolism and storage of the three major nutrients sugar, fat and protein. Insulin has two main effects on glucose metabolism: one is to promote the utilization of exogenous glucose, including promoting glucose transport, promoting glucose phosphorylation, promoting glycolysis, and promoting glucose oxidation; but inhibiting endogenous glucose production, including promoting glucose original synthesis, inhibit gluconeogenesis, and prevent liver glycogenolysis ^[37] . Insulin exists in the form of proinsulin, which is composed of C-peptide and the A and B chains of insulin. Proinsulin will generate a molecule of insulin and a molecule of C-peptide through the catalysis of protease and carboxypeptidase in the pancreas. Therefore, C-peptide and insulin are secreted by equal molecules, so the determination of insulin and C-peptide can directly reflect the function of pancreatic β-cells. Since insulin levels are affected by many factors, while C-peptide is relatively stable, the determination of C-peptide levels can accurately reflect the function of pancreatic β-cells. The experimental results of the present invention show that there is no significant difference between the insulin levels among the groups, which may be related to its instability; and fatty oil can significantly increase the C-peptide level of diabetic mice. It shows that fatty oil may play a hypoglycemic effect by promoting the secretion of insulin.

Insufficient secretion of insulin in diabetes can often cause abnormal lipid metabolism, because insulin can promote the decomposition of lipoproteins. When insulin secretion is insufficient or insulin resistance occurs in the body, the total cholesterol and triglycerides in the blood will increase significantly. Dyslipidemia, leading to diseases of the cardiovascular system. The experimental results showed that the fatty oil components could significantly reduce the levels of TC and TG in diabetic mice, suggesting that fatty oils can also regulate lipid metabolism while lowering blood sugar. the

Mitochondria in liver cells are the center of the body's energy supply and an important organ for maintaining blood sugar stability. Insulin signaling in the liver is of great significance to the maintenance of systemic blood sugar homeostasis and the normal function of the liver itself. On the one hand, diabetic patients mobilize a large amount of fat from the fat depot to the liver due to obstacles to the utilization of sugar. On the other hand, the insulin sensitivity of adipocytes decreases, and the ability to inhibit fat decomposition is weakened. During fat mobilization, visceral tissue mobilizes far more fat than subcutaneous fat, and excessive fat enters the liver, exceeding the ability of liver mitochondria to oxidize and decompose and synthesize lipoproteins to export outside the liver, causing excess fat to accumulate in liver cells. At the same time, hyperlipidemia can also lead to fat accumulation, infiltration and degeneration of liver tissue. The higher than normal concentration of sugar and lipid levels in the liver caused by diabetes affects the structure and function of organelles in liver cells. This pathological change in the liver affects the normal function of the liver, which will aggravate blood lipid metabolism and lipoprotein synthesis disorders. Trigger other energy metabolism disorders, forming a vicious circle. This experimental study found that Morus alba fatty oil can improve liver damage and has a better protective effect on liver cells in diabetic mice. the

Based on the above experimental results, the applicant found for the first time that Morus alba fatty oil can significantly improve blood sugar levels in diabetic mice, regulate glucose and lipid metabolism, and protect liver structure and function. The preparation method of Morus Alba Fatty Oil is simple, and after many experiments, the same or similar results have been obtained. In the past, no obvious medicinal value was found in the Morus Alba Fatty Oil, and it was generally discarded as a non-medicinal part. The research result of the present invention enables the waste to be utilized, expands medicinal resources, and opens up a new medicinal use of the Morus alba fatty oil. In addition, the Morus alba fatty oil has less dosage, is convenient to take, and has great clinical significance. the

Claims (4)

1. a Cortex Mori fatty oil is preparing the application in hypoglycemic medicine, this Cortex Mori fatty oil is not had Cortex Mori water logging, decocts 30-40min, removing medicinal residues, obtain Cortex Mori water extract, Cortex Mori water extract is concentrated, adds the petroleum ether extraction of 2 times amount, petroleum ether layer concentrates, after recycling design, vacuum drying 6h, obtains Cortex Mori fatty oil.

2. Cortex Mori fatty oil according to claim 1 is preparing the application in hypoglycemic medicine, it is characterized in that, described Cortex Mori fatty oil is, Cortex Mori 100g water logging is not had, decoct 30min, removing medicinal residues, obtain Cortex Mori water extract, Cortex Mori water extract is concentrated, add the petroleum ether extraction of 2 times amount, petroleum ether layer concentrates, after recycling design, vacuum drying 6h, obtains Cortex Mori fatty oil.

3. Cortex Mori fatty oil according to claim 1 is preparing the application in hypoglycemic medicine, it is characterized in that, described Cortex Mori fatty oil is, Cortex Mori 500g water logging is not had, decoct 35min, removing medicinal residues, obtain Cortex Mori water extract, Cortex Mori water extract is concentrated, add the petroleum ether extraction of 2 times amount, petroleum ether layer concentrates, after recycling design, vacuum drying 6h, obtains Cortex Mori fatty oil.

4. Cortex Mori fatty oil according to claim 1 is preparing the application in hypoglycemic medicine, it is characterized in that, described Cortex Mori fatty oil is, Cortex Mori 1000g water logging is not had, decoct 40min, removing medicinal residues, obtain Cortex Mori water extract, Cortex Mori water extract is concentrated, add the petroleum ether extraction of 2 times amount, petroleum ether layer concentrates, after recycling design, vacuum drying 6h, obtains Cortex Mori fatty oil.