CN100444849C - New use of tribulus terrestris extraction - Google Patents

Abstract

The present invention discloses an application of tribulus terrestris extract to the preparation of medicines, health-care products or food for treating or preventing depression. The present invention researches the depression resistance activity of tribulus terrestris extract, particularly the total saponin of the tribulus terrestris extract by rat depression models. Results indicate that the tribulus terrestris extract can obviously improve the depressive state of rats. The effective active constituent of the tribulus terrestris extract is total saponin, wherein every 1kg of the tribulus terrestris extract of a high dosage group comprises 200 mg of tribulus terrestris total saponin, the extract can obviously improve the contents of 5-HT and 5-HIAA in the brains of the rats, and flavone components and polysaccharide components in tribulus terrestris do not have depression resistance activity.

Description

New uses of Tribulus terrestris extract

technical field

The invention relates to a new application of the extract of Tribulus terrestris.

Background technique

The traditional Chinese medicine Tribulus terrestris is the dry mature fruit of Tribulus terrestris (Tritulus terrestris) in the family Zygophyllaceae, also known as Tribulus terrestris, Tribulus terrestris, Tribulus terrestris, and Tribulus terrestris. Pungent in nature and flavor, bitter, slightly warm, slightly poisonous, enters the liver and lung meridians, has the effects of dispersing wind, improving eyesight, lowering qi, promoting blood circulation, tonifying kidney and replenishing essence. Tribulus terrestris has a long history of medicinal use among Chinese folks. "Shen Nong's Materia Medica" listed "Tribulus terrestris" as a top-grade medicine, saying that it "maintains bad blood, breaks accumulation of crux, sore throat, breast difficulty, long-term use to grow muscles, and improve eyesight." , light body ". The Chinese Pharmacopoeia records its function as "calming the liver and relieving depression, promoting blood circulation and expelling wind, improving eyesight, and relieving itching. It is used for headache, dizziness, distending pain in the chest and hypochondrium, mastitis, red eyes and nebula, rubella and itching".

In recent years, the research on this plant has not been done in various countries. Studies have shown that Tribulus terrestris extracts include tribulus terrestris steroidal saponins (saponins), flavonoids, polysaccharides and alkaloids, among which, the main components can be extracted and separated by different methods to be saponins (Jin Jingling et al. Macroporous adsorption resin method to extract total tribulus terrestris Saponins. Yanbian University Medical Journal. March, 1999.22 (1): 29-31), flavonoids (Shiqin et al. Determination of 3 kinds of hydrolyzed flavonoid aglycones in Tribulus terrestris by reversed-phase high-performance liquid chromatography. Impurities in drug analysis; 1999, 19(2): 75~78 or Wu Qiongshi, etc. Determination of total flavonoids in Tribulus terrestris. Journal of Guangdong Pharmaceutical University; 1998, 14(4): 273~275) or polysaccharides (Shi Qin, etc. Colorimetric determination of polysaccharide content in Tribulus terrestris. China Pharmaceutical University; 1997, 28 (5): 291 ~ 293) Tribulus terrestris extract. Foreign countries have developed preparations such as "Tribustan" and "Vitanone" for enhancing sexual function and "Tribusaponin" for softening blood vessels using Tribulus terrestris as raw materials. Cardiovascular diseases. But so far there is no report about the treatment of depression with Tribulus terrestris extract.

Contents of the invention

The technical problem to be solved by the present invention is the above-mentioned problem, and a new application of the extract of Tribulus terrestris is provided, that is, the application of the extract of Tribulus terrestris in the preparation of medicine, health food or food for treating or preventing depression.

According to the present invention, the term Tribulus terrestris refers to the fruit of Tribulus terrestris in the same Chinese Pharmacopoeia. The Tribulus terrestris extract of the present invention refers to various crude extracts extracted from the fruit of the plant Tribulus terrestris (after extracting with water or ethanol, etc., no further separation and purification are required, including steroidal saponins, flavonoids, polysaccharides, etc. sugars and alkaloids, etc.) or Tribulus terrestris steroidal saponin extracts of various purity (the above crude extracts are obtained by various methods of separating and purifying steroidal saponins); among them, the total steroidal saponins The Tribulus terrestris extract with a weight content of ≥50% is also referred to as the total tribulus saponins in the present invention, which contains the total spirosteroid saponins and the total furostanol saponins of Tribulus terrestris.

According to the present invention, the preferred Tribulus terrestris extract is the total saponins of Tribulus terrestris.

Among them, the active ingredients of the above-mentioned medicines may contain other effective substances for treating or preventing depression in addition to Tribulus terrestris extract or its total saponins, such as various drugs currently used for antidepressants, such as fluoxetine, paroxetine, etc. Or sertraline etc.

The medicine or health product can be tablet, granule, pill, powder, capsule, oral liquid, suspension or emulsion and the like.

The positive and progressive effect of the present invention lies in that the present invention studies the antidepressant activity of the Tribulus terrestris extract through a rat depression model, and measures the content of neurotransmitters in the rat brain. The results show that Tribulus terrestris extract can significantly improve the depression state of rats, and wherein the high-dose group Tribulus terrestris total saponins (200mg/kg) can also significantly improve 5-HT (5-hydroxytryptamine) content and 5-HIAA ( 5-hydroxyindoleacetic acid) content, so its effective active ingredients are total saponins. Therefore, the Tribulus terrestris extract of the present invention, especially the total saponins of Tribulus terrestris can be used to prepare antidepressant drugs, health food or food, and can also be used for auxiliary treatment of patients with depression.

Detailed ways

The present invention will be further described below with examples and effect examples. Wherein the crude extract of Tribulus terrestris and the total saponins of Tribulus terrestris can be commercially available products.

The preparation of embodiment 1 Tribulus terrestris crude extract

Take 1 kg of the fruit of Tribulus terrestris as the raw material, reflux with five times of water to extract 3 times, each time for 1 hour, and the extract is spray-dried to obtain the crude extract of Tribulus terrestris. After determination, the crude extract of Tribulus terrestris contains 20% of total saponins (vanillin-perchloric acid method), 2% of polysaccharides and 1% of total flavonoids.

The preparation of embodiment 2 Tribulus terrestris total saponins

Take 1 kg of the raw material Tribulus terrestris, extract 3 times with eight times the amount of 75% ethanol under reflux, each time for 2.5 hours, combine the extracts, filter, place the filtrate in a cool place (temperature less than 10°C) for 12 hours, and centrifuge at 4000 rpm for 5 minutes Finally, take the supernatant, recover ethanol, concentrate to dry extract, dissolve into a solution with water and pass through a macroporous adsorption resin column (HPD100). After being applied to the column, wash with water until colorless, then wash with 30% ethanol until colorless, then elute with 70% ethanol, collect the eluate, recover the ethanol until it has no alcohol smell, spray dry it into a dry powder, and obtain the thorn The total saponins of Tribulus terrestris have a purity of 60%.

The preparation of embodiment 3 tribulus terrestris total flavonoids

Tribulus terrestris fruit 1kg, reflux extraction with 5 times the amount of water for 3 times, precipitate with ethanol, filter the precipitate, put the filtrate on a polyamide column, elute with water until colorless, then elute with acetone, collect the acetone eluate, Evaporate the acetone and dry to obtain the total flavonoids of the fruit of Tribulus terrestris. Content determination refers to the above-mentioned literature (Shiqin et al. Determination of 3 kinds of hydrolyzed flavonoid aglycones in Tribulus terrestris by reversed-phase high performance liquid chromatography. Drug Analysis Impurities; 1999, 19(2): 75-78 and Wu Qiongshi et al. Tribulus terrestris Determination of the content of total flavonoids in grass. Journal of Guangdong Pharmaceutical University; 1998, 14(4): 273-275), and its purity was determined to be 52%.

The preparation of embodiment 4 tribulus terrestris polysaccharide

It was prepared according to the above literature (Shiqin et al. Determination of polysaccharide content in Tribulus terrestris by colorimetry. Journal of China Pharmaceutical University; 1997, 28(5): 291-293).

1 kg of Tribulus terrestris fruit was reflux extracted with 95% ethanol, and the residue was decocted 3 times with 3 times the amount of water for 1 hour each time, and the water extract was combined. Take 1500 mL of water extract, add 300 ml of chloroform to the extract. Take 1500mL of water extract, add 300mL of chloroform, 100mL of n-butanol, stir magnetically for 10-15h, put it in a separatory funnel, let it stand to separate layers, and remove the chloroform layer (containing protein). Then put the water layer in a dialysis bag and dialyze for 72h (to remove small molecular impurities and organic solvents). After the dialysis is finished, the dialysate is freeze-dried to obtain refined polysaccharides from Tribulus terrestris.

Effect Example 1 Antidepressant effect of Tribulus terrestris extract and total saponins of Tribulus terrestris on depression model rats-forced swimming test

①Modeling: Wistar rats (200-220g) were placed in a glass cylinder with an inner diameter of 20cm and a height of 40cm, one for each cylinder, the water depth in the cylinder was 24em, and the water temperature was 28°C-29°C (the water in each cylinder was only used once) . On the first day of the experiment, each rat swam for 15 minutes, then took it out, dried it under the lamp, put it in a cage, and tested it 24 hours later. The rats were put back into the water tank and observed continuously for 5 minutes. The accumulated rats stopped struggling in the water, were in a floating and upright state, and had only occasional limb movements to keep their heads floating on the water surface (immobilization time) for ten consecutive days. Six days of forced swimming. Those who did not move for more than 150s were defined as depression model animals.

② intragastric administration: with the crude extract of Tribulus terrestris in Example 1 and the total saponins of Tribulus terrestris in Example 2 as samples, divide into high-dose group (200mg/kg) and low-dose group (100mg/kg); Qulin (produced by Shanghai Institute of Pharmaceutical Industry) was used as a control group (4.5 mg/kg); normal saline was used as a blank control (5 ml/body). Each rat was administered 3 times, 24h, 5h, and 1h before the determination of immobility time, orally administered.

③ Swimming test: After intragastric administration, the forced swimming test was carried out according to the steps in ①, and the swimming immobility time of each wistar rat was recorded within 5 minutes. The results are shown in Table 1.

④Monitoring: After swimming, the rats were sacrificed, the brain was taken out, and the monoamine neurotransmitters in the rat brain were measured by fluorescence method, 5-hydroxytryptamine (5-HT) (provided by sigma), 5-hydroxyindoleacetic acid (5- HIAA) (provided by sigma), dopamine (DA) (provided by sigma) and norepinephrine (NA) (provided by sigma), the results are shown in Table 2.

Table 1 immobility time of forced swimming of wistar rats

^* p<0.01 compared with blank group

Table 2 Determination of neurotransmitters in wistar rat brain by fluorescence method

Compared with the blank group, ^* p<0.01, ^** p<0.05

The comparison of effect embodiment 2 antidepressant effect

Get the ICR mice (20-22g), the total saponins of Tribulus terrestris (143.6mg/kg), the polysaccharides of Tribulus terrestris (14.4mg/kg), the total flavonoids of Tribulus terrestris ( 7.2mg/kg) as the sample, sertraline as the control group (5.92mg/kg), and normal saline as the blank control (0.5ml/cat), and they were administered by gavage for 14 consecutive days. After gavage on the 14th day, they were forced to swim for 6 minutes , after calculating the swimming immobility time within 4 minutes, the results are shown in Table 3. Wherein, the ratio of saponins, polysaccharides and flavonoids contained in the crude extract of Tribulus terrestris extracted in Example 1 is about 10:1:0.5, and the dosage ratio of the above three in the test is set.

Table 3 The immobility time of forced swimming of ICR mice

^* p<0.01 compared with blank group

The above effect examples 1 and 2 studied the antidepressant activity of the extract of Tribulus terrestris through the rat depression model, and measured the content of neurotransmitters in the rat brain. The results show that Tribulus terrestris extract can significantly improve the depression state of rats, and wherein the high-dose group Tribulus terrestris total saponins (200mg/kg) can also significantly improve 5-HT (5-hydroxytryptamine) content and 5-HIAA ( 5-hydroxyindoleacetic acid), but has no effect on the content of DA and NE, and its effect is similar to that of the third class of antidepressant sertraline. Further studies on the antidepressant activity of flavonoids and polysaccharides in Tribulus terrestris showed that neither of them has antidepressant effect.

Claims (1)

1, the purposes of tribulus terrestris extraction in medicine, health food or the food of preparation treatment or prevention depression is characterized in that this tribulus terrestris extraction is a Fructus Tribulus total saponins.