CN112121042A - Application of PSO in preparation of anti-septicemia and myocardial damage drug induced by anti-septicemia - Google Patents

Abstract

The present invention discloses the application of psoralen for preparing medicine for treating and/or preventing sepsis and myocardial injury induced by it. The research of the present invention found that PSO can improve the survival rate of sepsis, improve the score of sepsis in mice, reduce the number of white blood cells, lymphocytes, monocytes, granulocytes, and reduce the levels of lactate dehydrogenase, urea nitrogen, creatine kinase and aspartate aminotransferase ; PSO can improve cardiac systolic function and maintain the normal shape of myocardial tissue; in addition, PSO can also inhibit the high expression of myeloperoxidase and interleukin-1β after sepsis-induced myocardial injury, reduce the inflammation-related molecule IL-1β , IL-6, NOD-like receptor family 3 mRNA levels, and reduce superoxide anion accumulation caused by sepsis, thereby exerting anti-inflammatory and anti-oxidative stress effects.

Description

Application of PSO in the preparation of anti-septicemia and its induced myocardial injury drugs

technical field

The invention relates to a new indication of Psoralidin (PSO), in particular to the application of PSO as an anti-septicemia medicine and the induced myocardial injury medicine.

Background technique

Psoralen PSO has the structural formula shown in formula I, and it has various pharmacological activities, including antioxidant, antibacterial, anti-inflammatory, antidepressant and estrogen-like effects.

SUMMARY OF THE INVENTION

By constructing sepsis and its induced myocardial injury animal model, the inventors observed various indicators of survival rate, blood routine, blood biochemistry, inflammation, oxidative stress and myocardial injury and found:

PSO can improve sepsis survival rate; improve sepsis score; reduce white blood cell (WBC), lymphocyte (Lymphocyte, LYM), monocyte (MON), granulocyte (Granulocyte, GRA) number and reduce lactate Levels of Lactate Dehydrogenase (LDH), Blood Urea Nitrogen (BUN), Creatine Kinase (CK), and Aspartate Aminotransferase (AST);

At the same time, PSO can improve myocardial tissue damage and cardiac systolic function caused by sepsis, at least by increasing the left ventricular end-systolic volume (Volume; s) and the decrease in left ventricular end-diastolic volume (Volume; d); and reducing left ventricular contraction End-stage posterior wall thickness (LVPW; s) and left ventricular end-diastolic posterior wall thickness (LVPW; d) and heart rate (Heart Rate, HR) increase, so as to improve cardiac systolic function;

PSO can also inhibit the high expression of Myeloperoxidase (MPO), Interleukin-1β (IL-1β), Ly6c, F4/80 after sepsis-induced myocardial tissue injury, reduce IL-1β , IL-6, NOD-like receptor pyrin domain-containing protein 3 (NLRP3) mRNA levels, and reduce superoxide anion accumulation caused by sepsis, thereby exerting anti-inflammatory and anti-oxidative stress effect;

Based on the above findings, the present invention provides the application of PSO for preparing a medicine for treating and/or preventing sepsis and its induced myocardial damage.

At the same time, a medicine for treating and/or preventing sepsis is provided, which is prepared from PSO and pharmaceutical excipients.

Further, the drug is a preparation for intravenous injection.

Further, the dosage of the drug is 12.5mg-50mg PSO per kilogram of body weight.

Description of drawings:

Figure 1 shows the effect of PSO on the survival rate of mice after CLP injury. The survival of mice in each group within 72 hours after CLP was observed. Picture A is the modeling picture of the survival rate of mice, and picture B is the survival rate curve of mice. Results Expressed as mean±standard deviation, n=12; ^* vs. Sham group, P<0.05;^# vs. CLP group, P<0.05;

Figure 2 shows the effect of PSO on the septicemia score of mice after CLP injury. At 8 hours after the operation, the mice were scored according to the relevant indicators of the septicemia score. The picture of the mice after the injury is shown in picture A, the model picture of the functional test of mice, picture B is the result of sepsis score, the results are expressed as mean ± standard deviation, n=8; ^* vs. Sham group, P<0.05;^# vs. CLP group;

Figure 3 shows the effect of PSO on various indicators of blood routine in mice after CLP injury. Blood routine was measured 8 hours after operation in mice, and the results were expressed as mean ± standard deviation, n=6; ^* vs. Sham group, P<0.05 ; ^# vs. CLP group;

Figure 4 shows the effect of PSO on the blood biochemical indexes of mice 8 hours after CLP injury. The blood routine was measured 8 hours after the operation. The results were expressed as mean ± standard deviation, n=6; ^* vs. Sham group, P<0.05;^# vs. CLP group;

Figure 5 shows the effect of PSO on the morphology of myocardial tissue 8h after CLP injury, the results of HE staining of myocardial tissue sections; HE, hematoxylin-eosin;

Figure 6 shows the effects of PSO on various indicators of cardiac function 8 hours after CLP injury. The mice were detected by echocardiography 8 hours after operation. Picture A is the long-axis section of the parasternal left ventricle of the echocardiogram, typical pictures of the M model, and various Statistical analysis chart of cardiac function indicators, picture B is the short-axis section of echocardiography, typical pictures of M mode and statistical analysis chart of various cardiac function indicators, the results are expressed as mean ± standard deviation, n=6, ^* vs. Sham group, P<0.05;^# vs. CLP group, P<0.05; left ventricular end-systolic volume; left ventricular end-diastolic volume; left ventricular end-systolic posterior wall thickness; left ventricular end-diastolic posterior wall thickness; heart rate;

Figure 7 shows the effect of PSO on myocardial inflammation-related indicators 8 hours after CLP injury, the immunohistochemical staining results and statistical analysis of typical inflammation-related indicators MPO and IL-1β; the mRNA expressions of inflammation-related molecules IL-1β, IL-6 and NLRP3 Among them, Panel A is the result of immunohistochemical staining, Panel B is the result of mRNA expression level, the results are expressed as mean ± standard deviation, n=6, ^* vs. Sham group, P<0.05;^# vs. CLP group, P<0.05;

Figure 8 shows the effect of PSO on the level of superoxide anion in myocardial tissue after 8 h of CLP injury, the results of DHE staining and statistical analysis of myocardial tissue, the results are expressed as mean ± standard deviation, n=6. ^* vs. Sham group, P<0.05;^# vs. CLP group, P<0.05, DHE, superoxide anion fluorescent probe.

Detailed ways

Sepsis refers to a systemic infection syndrome in which various pathogenic bacteria invade the blood circulation and rapidly grow and multiply in the blood, producing a large number of toxins and metabolites, causing severe toxemia. The aggravation of sepsis can cause septic shock, disseminated intravascular coagulation and organ function impairment, which seriously threatens the life of patients. The heart is one of the target organs vulnerable to sepsis, and there is ample evidence that sepsis can lead to cardiac dysfunction. Clinical studies have shown that the mortality rate of patients with sepsis complicated with cardiac dysfunction is as high as 70%-90%. In contrast, patients with sepsis without cardiac dysfunction have a mortality rate of approximately 20%.

Cecal ligation and puncture (Cecal Ligation and Puncture, CLP) animal model is a classic sepsis animal model, the present invention adopts CLP animal model as research object, according to prior art research, this animal model can cause acute myocardial injury, such as sepsis-induced myocardial injury damage.

Myocardial injury induced by sepsis is a type of infectious myocardial injury. Infectious myocardial injury refers to reactions such as cardiac enlargement, heart failure, cardiogenic shock, or abnormal heart rhythm that occur during viral infection or during recovery. Typical symptoms are fatigue. Weakness, loss of appetite, nausea, vomiting, difficulty breathing, paleness, and fever.

Infectious myocardial injury in the present invention particularly refers to bacterial endocarditis, myocarditis lesions caused by bacteremia caused by systemic or other organ infection, and sepsis caused by systemic or other organ infection In order to prevent myocarditis caused by the disease, adrenal corticosteroids and broad-spectrum antibiotics are often used clinically to prevent dysbacteriosis. Strict observation should be made during the application, and special attention should be paid to the presence or absence of fungal infection of the digestive tract, urinary tract and respiratory tract.

The present invention will be further described below through examples. Examples are given to describe the present invention in detail, but the present invention is not limited to these at all.

It should be noted that the purity of PSO used in the following examples is HPLC≥98%. The animals used were purchased from the Experimental Animal Center of the Fourth Military Medical University, and the reagents used were purchased from the market. Unless otherwise specified, the experimental methods or related detection methods used in the following examples adopt methods known in the art.

Example 1: The inventor's study found that PSO can improve the death of mice caused by sepsis

Program:

Using CLP surgery, the sepsis model was constructed at the in vivo level, and PSO pre-protection was given.

step:

Wild-type BALB/c mice were used as the research objects, according to the research design, grouped by the random number table method, and the model of infective myocardial injury in mice was replicated according to the CLP experimental method published by Rittirsch D et al. The specific experimental steps are as follows:

(1) Grouping: Survival rate experiment: BALB/c mice were divided into sham operation group, CLP group, and PSO group (12.5, 25, and 50 mg/kg doses), with 12 mice in each group;

(2) Administration: Pre-protection treatment (intraperitoneal injection) was performed 6 days before the operation, and DMSO (dimethyl sulfoxide) was administered to the sham-operated group and CLP group at a concentration of 1 ml/kg (body weight), and the protection group PSO (DMSO dissolved, PSO solution with doses of 12.5mg/kg, 25mg/kg and 50mg/kg respectively), administered once every 2 days for a total of 3 times, to ensure that the time period of each administration and the operation time are the same;

(3) Model establishment: ① Model establishment on the 7th day, the mice were anesthetized with a small animal inhalation anesthesia system: the mice inhaled oxygen containing isoflurane 2% (volume fraction vol/vol), and the anesthesia flow was 3L/min , during the modeling process, the flow rate was 1.5L/min, and the monitoring standard for the degree of anesthesia was the disappearance of the withdrawal reflex of the limbs. The mice were immobilized and continuously inhaled with 2% oxygen containing isoflurane to maintain anesthesia; The skin was disinfected twice with 75% ethanol, a 1 cm longitudinal incision was made along the middle of the lower abdomen, and the skin and subcutaneous tissue were incised layer by layer to separate the skin and subcutaneous tissue. Moisten with % normal saline, enter the abdomen with curved forceps, find the cecum and pull it out gently, gently squeeze the feces near the ileocecal valve to the end of the cecum (to avoid air remaining), and 2/3 of the line connecting the end of the cecum to the ileocecal valve (The survival rate experiment is this aggravated model, and the functional test adopts the ligation model at the 1/3 of the connection between the end of the cecum and the ileocecal valve, and the rest of the steps are the same as the aggravated model) ligature the cecum with No. 4 sterile surgical suture, and then ligate the cecum. Use a 2.5ml syringe needle to pierce the ligated intestine (avoiding blood vessels) at the midpoint of the end of the line and the cecum. After perforation, gently squeeze the cecum. It can be seen that the contents of the cecum in the ligated segment flow out through the puncture hole, and the cecum and all surrounding intestinal tubes are also included. In the abdominal cavity, the peritoneum and skin were sutured layer by layer with No. 3 sterile surgical sutures. Put it back in the cage and wait for it to wake up. In the sham operation group, the rest of the steps were the same as in the experimental group except that the cecal ligation and perforation were not performed.

(4) Start timing after CLP operation, observe every 1 h, record the number of deaths of mice in each group within 72 h, and count and analyze the survival rate;

(5) According to the survival rate results of each group, after selecting the appropriate concentration of PSO under the scheme of this embodiment, the animal experiment is carried out as above, and the obtained specimens are subjected to subsequent detection.

result:

The survival rate curve of mice is shown in Figure 1B. Compared with the control group, the survival rate of mice within 72 hours after CLP treatment was 40% (P<0.05), and the survival rate of mice treated with PSO (12.5 mg/kg) was about 40% (P<0.05). was 70% (P<0.05), the survival rate of mice after PSO (25mg/kg) treatment was about 70% (P<0.05), and after PSO (50mg/kg) treatment, the survival rate of mice was about 80% (P <0.05), suggesting that PSO can improve the survival rate of mice after CLP, and the optimal protective concentration of PSO is 50 mg/kg (the optimal protective concentration will be used in subsequent functional tests).

Example 2: The inventor's research found that PSO can improve the sepsis score and blood routine changes caused by sepsis.

Program:

Using CLP surgery, a model of sepsis and its induced myocardial injury was constructed at the body level (all subsequent functional tests were performed by ligating the 1/3 of the line connecting the end of the cecum to the ileocecal valve, and the rest of the steps were the same as the aggravation model, see Figure 2A ), and give PSO processing.

step:

(1) The mice at 8 h after CLP were scored according to the sepsis score table, and the scoring rules adopted the content disclosed in the following documents:

[1] Nemzek JA, Hugunin KM, Opp MR: Modeling sepsis in the laboratory: merging sound science with animal well-being. Comp Med 2008, 58:120–128.

[2] Huet O, Ramsey D, Miljavec S, Jenney A, Aubron C, Aprico A, Stefanovic N, Balkau B, Head GA, de Haan JB, Chin-Dusting JP: Ensuring animal welfare while meeting scientific aims using a murine pneumonia model of septic shock. Shock (Augusta, Ga) 2013, 39:488–494.

[3] Langford DJ, Bailey AL, Chanda ML, Clarke SE, Drummond TE, Echols S, Glick S, Ingrao J, Klassen-Ross T, Lacroix-Fralish ML, Matsumiya L, Sorge RE, Sotocinal SG, Tabaka JM, Wong D, van den Maagdenberg AM, Ferrari MD, Craig KD, Mogil JS: Coding of facial expressions of pain in the laboratory mouse. Naturemethods 2010, 7:447–449.

[4] Bradly Shrum, Ram V Anantha, Stacey X Xu, Marisa Donnelly, SM Mansour, Haeryfar, John K McCormick: A robust scoring system to evaluate sepsis severity in an animal model. BMC Research Notes 2014, 7:233.

(2) Detect the changes of blood routine indexes of mice after 8 hours of CLP operation: After 8 hours of injury, the blood was collected by removing the eyeball method, and the blood routine test was performed by an automatic blood routine instrument.

result:

The mice were treated with CLP for 8 hours for sepsis score. The results are shown in Figure 2. Compared with the control group, the sepsis score in the CLP group was significantly increased, and after PSO protection, the sepsis score was significantly reduced;

The changes of blood routine related indexes were detected after 8 h of CLP treatment in mice. As shown in Figure 3, compared with the control group, WBC and LYM increased significantly (P<0.05), while WBC and LYM decreased significantly after PSO treatment (P<0.05). P<0.05), but had no significant effect on RBC and PLT (P>0.05).

Example 3: The inventor's research found that PSO can improve blood biochemical changes caused by sepsis

Program:

Using CLP operation, the sepsis and its induced myocardial injury model were constructed at the body level, and PSO treatment was given.

step:

Detect the changes of blood biochemical indexes of mice after 8 hours of CLP operation: After 8 hours of injury, blood was collected by removing the eyeball method, collecting whole blood from each group, centrifuging at 3000 r/min for 10 min, drawing serum, and then using automatic blood biochemistry analyzer to detect.

result:

The changes of blood biochemical indexes were detected after 8 hours of CLP treatment in mice. As shown in Figure 4, compared with the control group, after 8 hours of CLP injury, the levels of LDH, BUN, CK and AST in serum were significantly increased (P<0.05). ), after PSO treatment, the levels of LDH, BUN, CK and AST decreased significantly (P<0.05).

Example 4: The inventor's research found that PSO can improve myocardial tissue damage caused by sepsis.

Program:

Using CLP operation, the sepsis and its induced myocardial injury model were constructed at the body level, and PSO treatment was given.

step:

(1) HE staining of myocardial tissue:

① Paraffin embedding: Slowly inject heparin-containing normal saline into the apex of the heart, and when the fluid flowing out of the right atrial appendage becomes transparent, replace the perfusion fluid with 4% paraformaldehyde fixative; Blood vessels, the heart is completely removed; after the heart is removed, the left half of the heart (left atrium and left ventricle) is cut off and placed in 4% paraformaldehyde, post-fixed for at least 24 hours; 80%, 95%, 95% in order , soaked in 100% ethanol for 40 min, and then soaked in 100% ethanol, 100% ethanol: xylene = 1:1 mixture, and xylene concentration gradient for 30 min to dehydrate the tissue; then immersed in wax for 3 h in the embedding machine, and finally Drop wax embedding.

② Slicing: Set the slice thickness to 5 μm, stick the slice on the poly-lysine-coated glass slide using the pick-up method, bake the slice at 70°C for 1 hour, and bake it at 60°C for 5 hours.

③Staining: Immerse the sections in xylene for 10 min, replace the xylene and soak again for 10 min, soak in the order of 100%, 100%, 95%, 95%, 80% ethanol, and deionized water for 2 min to deparaffinize to water. For staining; immerse the sections in hematoxylin staining solution for 3 min, and gently rinse with tap water for 5 min; immerse in 1% hydrochloric acid ethanol for 30 s, 1% ammonia water for 3 min for decolorization, and gently rinse with tap water for 3 min; immerse in eosin staining solution for 3 min and gently rinse with tap water 3min; soak in 70%, 80% concentration ethanol for 30s, 95%, 95%, 95%, 100%, 100% concentration gradient ethanol, xylene, xylene for 2min respectively for dehydration and transparent treatment; neutral resin seal .

result:

HE staining results of mouse myocardial tissue showed that, as shown in Figure 5, compared with the control group, the myocardial tissue structure was disordered after CLP injury, the number of nuclei in the visual field increased significantly, and the infiltration of microvascular mononuclear cells between muscle cells increased; Compared with PSO, HE staining showed that the myocardial tissue structure was relatively clear, the number of nuclei in the visual field was reduced, and the infiltration of microvascular mononuclear cells between muscle cells was reduced.

Example 5: The inventor's study found that PSO can improve myocardial function damage caused by sepsis

Program:

Using CLP operation, the sepsis and its induced myocardial injury model were constructed at the body level, and PSO treatment was given.

step:

Ultrasound of small animals to detect the cardiac function of mice 8 hours after CLP operation: The animals in each group took off the coat on the left thoracic region of the mice one day before the ultrasound examination, and the mice were anesthetized with 2% isoflurane, the anesthetic gas flow was 1 L/min, and the isoflurane was anesthetized. After halothane inhalation anesthesia, the patients were fixed on a 37°C thermostatic plate, and the left thorax was fully exposed. A 30MHz probe was used to select the standard parasternal long-axis view of the left ventricle and the standard short-axis view of the left ventricular papillary muscle, and record the M-mode echocardiographic images. Measurement indicators include: left ventricular end-systolic volume, left ventricular end-diastolic volume, left ventricular end-systolic posterior wall thickness, left ventricular end-diastolic posterior wall thickness, heart rate, etc.

result:

Small animal ultrasound detected myocardial contractile function in mice 8 hours after CLP surgery. The left ventricular end-systolic volume and left ventricular end-diastolic volume were significantly decreased (P<0.05), and the cardiac function was significantly improved after PSO protection (P<0.05); The wall thickness and posterior wall thickness of the left ventricle at end-diastole were significantly thicker, and the heart rate was significantly increased (P<0.05). After PSO protection, the cardiac function was significantly improved (P<0.05).

Example 6: The inventor's research found that PSO can improve myocardial injury caused by infectious CLP by reducing inflammatory response.

Program:

Using CLP operation, the sepsis and its induced myocardial injury model were constructed at the body level, and PSO treatment was given.

step:

(1) Immunohistochemical detection:

①The steps of paraffin embedding and sectioning are the same as above;

②Staining: routine deparaffinization of sections to water: paraffin sections of mouse heart tissue in each group were taken and subjected to xylene twice, 10 minutes each time, 100% ethanol twice, each time 10 minutes; 95%, 90%, 80%, 70% ethanol once for 5 min each, and finally immersed in distilled water for 5 min; antigen retrieval: microwave antigen retrieval in sodium citrate buffer for 20 min, rinsed with running water for 10 min; blocking endogenous peroxidase: 3% hydrogen peroxide, room temperature for 15 min . Wash with PBS for 3 times, 5 min each time; block: add 5% normal goat serum blocking solution dropwise, incubate at room temperature for 60 min; drop primary antibody: wipe off excess serum, add primary antibody, and incubate overnight at 4°C. Wash 3 times with PBS, 5 min each time; add secondary antibody dropwise: add horseradish peroxidase HRP-labeled secondary antibody (1:5000, prepared in PBS), incubate for 1 h in a 37°C incubator, wash 3 times with PBS, each time DAB color development: add DAB dropwise for 0.5-3 minutes, control the degree of color development under the microscope, rinse with running water for 10 minutes, counterstain with hematoxylin, differentiate with 1% hydrochloric acid alcohol, decolorize with 1% ammonia water, dehydrate, clear with xylene and then use neutral gum seal.

③ Observe and take pictures under a microscope: Observe and take pictures under a microscope. The tissue sections with brownish-yellow granular deposition areas under light microscopes are positive staining sites. Randomly find 20-30 non-overlapping visual fields for each section, and use medical image analysis software. Image J 5.0 software semi-quantitative calculation.

(2) Real-time quantitative fluorescent PCR

①Extraction of total RNA from the sample: Take out the centrifuge tube containing the mouse heart sample from the liquid nitrogen, add 1 mL of Trizol and 2 grinding beads, place it on a tissue disrupter at 60 Hz for 1 min, and shake gently to make Trizol and the sample Mix well and place on ice to lyse for 5 min; add 200 μL of chloroform, shake the tube vigorously for 15 s, incubate at room temperature for 15 min, and centrifuge in a high-speed centrifuge at 4°C for 15 min at a speed of 12000 rmp/min; at this time, all RNA is in the upper aqueous phase Transfer 400 μL of the aqueous phase to a clean RNase-free centrifuge tube with a pipette, add an equal volume of isopropanol to mix, shake by hand for 15 s, incubate at room temperature for 10 min, and place it in a high-speed centrifuge at 4°C for 15 min. , the speed is 12000rmp/min; discard the supernatant, add 75% ethanol (prepared with DEPC water), shake manually to suspend the RNA precipitate, incubate at room temperature for 5min, centrifuge at 4°C for 5min at 8000rmp/min, repeat this operation once; discard ethanol solution, leaving the RNA precipitate, and drying at room temperature for 5-10 min; dissolve the RNA precipitate with 20 μL of DEPC water, turn on the DNA/RNA concentration analyzer, measure the concentration of the sample, and store it in a -80°C refrigerator.

②Reverse transcription: Take the extracted total RNA out of the -80°C refrigerator, add reverse transcription reagent (purchased from Hunan Aikerui Bioengineering Co., Ltd.; Item No.: AG11706), and place it in a PCR instrument for reverse transcription. The reverse transcription program is: 37 °C for 15 min; 85 °C for 5 s; the reversed cDNA is stored at -20 °C. The reverse transcription system is shown in Table 1:

Table 1 Reverse transcription system

③qRT-PCR: cDNA, kit (purchased from Hunan Aikerui Bioengineering Co., Ltd.; product number: AG11409-S) and the required primers (purchased from Shanghai Sangon Bioengineering Co., Ltd.; NLRP3: forward primer 5'- GGAGTTCTTCGCTGCTATGTA-3'; reverse primer 5'-GGACCTTCACGTCTCGGTTC-3'; IL-1β: forward primer 5'-GTGTCTTTCCCGTGGACCTT-3'; reverse primer 5'-CATCTCGGAGCCTGTAGTGC-3'; IL-6: forward primer 5 '-TTGGGACTGATGCTGGTGAC-3'; reverse primer 5'-GGTATAGACAGGTTCTGTTGGGAGT-3') was taken out from the -20°C refrigerator, and the equipment required for the experiment was prepared; after the sample and the kit were melted, the solution was prepared according to the following ratio, and then used The RT-qPCR instrument set the program for the experiment; the reaction conditions were: 95°C pre-denaturation for 10 min; 95°C denaturation for 15s; 58°C annealing and extension for 20s; 72°C final extension for 30s, a total of 40 cycles, the reaction system is shown in Table 2- 1, 2-2, 2-3 show:

Table 2-1 qRT-PCR reaction system

Table 2-2 qRT-PCR reaction system

Table 2-3 qRT-PCR reaction system

result:

The IHC staining results of mouse myocardial tissue showed that, as shown in Figure 7A, compared with the control group, the expressions of MPO and IL-1β were significantly increased after CLP injury, and their expressions were significantly decreased after PSO treatment;

The qRT-PCR results of mouse myocardial tissue showed that, as shown in Figure 7B, compared with the control group, the mRNA levels of inflammation-related molecules IL-1β, IL-6, and NLRP3 were significantly increased after CLP injury (P<0.05). The expression level decreased after PSO treatment (P<0.05).

Example 7: The inventor's study found that PSO ameliorated myocardial damage caused by infectious CLP by reducing superoxide anion accumulation caused by sepsis.

Program:

Using CLP operation, the sepsis and its induced myocardial injury model were constructed at the body level, and PSO treatment was given.

step:

DHE staining:

①The steps of paraffin embedding and sectioning are the same as above.

②DHE staining: soak the sections in xylene for 10 minutes, replace the xylene and soak for 10 minutes again, soak in 100%, 100%, 95%, 95%, 80% ethanol and deionized water for 2 minutes to deparaffinize to water, then use Prepare for staining; after dewaxing, use DHE staining solution to incubate at 37 °C for 30 min, after proper washing, observe and take pictures under a fluorescence microscope, the tissue sections under the fluorescence microscope show red fluorescence positive staining, and each section randomly finds 20 -30 non-overlapping visual fields were used to semi-quantitatively calculate the relative content of positive substances using the medical image analysis software Image-J 5.0 software.

result:

The DHE staining results of mouse myocardial tissue showed that, as shown in Figure 8, compared with the control group, the red fluorescence sites were significantly increased after CLP injury, and the red fluorescence intensity was significantly enhanced (P<0.05), and the fluorescence sites were significantly reduced after PSO treatment. , the fluorescence intensity decreased significantly (P<0.05).

Claims (5)

1. Use of PSO for the preparation of a medicament for the treatment and/or prevention of sepsis.
2. Use of PSO for the preparation of a medicament for the treatment and/or prevention of sepsis-induced myocardial injury.
3. 3. The medicine for treating and/or preventing septicemia and myocardial damage induced by septicemia is characterized by being prepared from PSO and pharmaceutical excipients.
4. 4. The medicament of claim 3, wherein the medicament is an intravenous formulation.
5. 5. The medicament of claim 3 or 4, wherein said medicament is administered in a dose of 12.5mg to 50mg PSO per kilogram body weight.

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