CN112972638B - Application of psoriasis treating traditional Chinese medicine in preparation of medicine for preventing and treating myocardial ischemia reperfusion injury - Google Patents

Abstract

The invention relates to an application of a traditional Chinese medicine for treating psoriasis in preparing a medicine for preventing and treating myocardial ischemia-reperfusion injury, wherein the effective components of the traditional Chinese medicine for treating psoriasis comprise powder prepared from 20 parts by weight of glabrous greenbrier rhizome and water extracts of the following raw material medicines: 10 parts of curcuma zedoary, 20 parts of glabrous sarcandra herb, 20 parts of lithospermum, 15 parts of red paeony root, 20 parts of dark plum fruit, 10 parts of liquorice, 10 parts of Chinese angelica, 10 parts of szechuan lovage rhizome and 20 parts of rehmannia root. The medicine for preventing and treating myocardial ischemia-reperfusion injury can improve the cardiac function of a model rat, inhibit inflammatory reaction, regulate immunity, relieve the pathological change of myocardial fibrosis and relieve myocardial ischemia-reperfusion injury.

Description

Application of psoriasis treating traditional Chinese medicine in preparation of medicine for preventing and treating myocardial ischemia reperfusion injury

Technical Field

The invention relates to a medicinal preparation with an undetermined structure containing traditional Chinese medicines, in particular to a medicament for treating cardiovascular system diseases.

Background

The morbidity and mortality of coronary heart disease myocardial infarction are rising year by year, and with the development and progress of medical research, various treatment means such as thrombolysis, percutaneous coronary intervention, coronary artery stenting and the like are generally applied in clinical treatment. While restoring myocardial blood supply and reducing myocardial damage through these therapies, ischemia-reperfusion may exacerbate cardiac dysfunction and structural damage and even cause irreversible death of cardiomyocytes, known as ischemia-reperfusion injury (MIRI). Therefore, much attention has been paid to prevention and treatment of myocardial ischemia-reperfusion injury, and it is a hot issue of research.

Ischemia reperfusion injury involves inflammatory reactions, the release of large amounts of free radicals, the activation of excitatory amino acids and metabolic receptors, the activation of endonucleases, the activation of platelet activating factor, etc. The body's stressful (immunological) inflammatory responses underlie ischemia reperfusion injury, and various inflammatory mediators, including cytotoxins, underlie these inflammatory responses.

The traditional Chinese medicine can intervene in the inflammatory reaction after myocardial ischemia/reperfusion injury by various ways of inhibiting chemotaxis and infiltration of inflammatory cells, regulating synthesis and secretion of cytokines, restoring the balance of inflammatory inhibitory factors and proinflammatory factors and the like, and plays a role in protecting the myocardium. In recent years, researches prove that the traditional Chinese medicine has the effects of inhibiting platelet activation, regulating inflammatory response, resisting thrombosis, inhibiting oxidative stress injury, promoting myocardial capillary vessel neogenesis and the like, can act on a plurality of pathological links of myocardial injury, and shows good application prospects.

Example 1 of the patent application with publication number CN105233150A discloses a traditional Chinese medicine composition, which is prepared from the following raw materials in parts by weight; 10 parts of curcuma zedoary, 20 parts of glabrous sarcandra herb, 20 parts of sinkiang arnebia root, 15 parts of red paeony root, 20 parts of glabrous greenbrier rhizome, 20 parts of dark plum fruit, 10 parts of liquoric root, 10 parts of Chinese angelica, 10 parts of szechuan lovage rhizome and 20 parts of rehmannia root. Meanwhile, the above patent application also discloses that the traditional Chinese medicine composition has the following effects and therapeutic uses: the invention has the functions of clearing away heat and toxic material, promoting blood circulation and removing blood stasis, is used for treating psoriasis, and has definite curative effect; has the functions of dispelling wind and clearing heat, eliminating dampness and reducing swelling, and promoting blood circulation and removing obstruction in channels, is used for treating psoriatic arthritis and rheumatoid arthritis, and has definite curative effect; the invention also has the functions of promoting diuresis and clearing heat, activating blood and dissolving stasis, and detoxifying and resolving masses, is used for treating malignant tumors, and has definite curative effect. However, there is no report on other new uses of the Chinese medicinal composition described in the above patent applications.

Disclosure of Invention

The invention aims to solve the technical problem of providing a new application of a traditional Chinese medicine for treating psoriasis in pharmacy.

The new application of the traditional Chinese medicine for treating psoriasis in pharmacy is specifically as follows:

the application of the traditional Chinese medicine for treating psoriasis in preparing the medicine for preventing and treating myocardial ischemia-reperfusion injury is characterized in that the effective components of the traditional Chinese medicine for treating psoriasis are the same as in example 1 of the patent application with the publication number of CN105233150A, namely the traditional Chinese medicine for treating psoriasis is composed of powder prepared from 20 parts by weight of glabrous greenbrier rhizome and water extracts of the following raw material medicines:

10 parts of curcuma zedoary, 20 parts of glabrous sarcandra herb, 20 parts of lithospermum, 15 parts of red paeony root, 20 parts of dark plum fruit, 10 parts of liquorice, 10 parts of Chinese angelica, 10 parts of szechuan lovage rhizome and 20 parts of rehmannia root.

In the application, the effective components are prepared by the following method:

(1) pulverizing rhizoma Smilacis Glabrae, and sieving with 80 mesh sieve to obtain rhizoma Smilacis Glabrae powder;

(2) selecting Curcumae rhizoma, herba Pileae Scriptae, radix Arnebiae, radix Paeoniae Rubra, mume fructus, and fructus PhyllanthiDecocting herbal pieces of grass, angelica, ligusticum wallichii and radix rehmanniae for three times, adding 10-12 times of water for the first time, decocting and extracting for 2 hours, and adding 8-10 times of water for the second time and the third time respectively, and decocting and extracting for 1 hour; mixing the extractive solutions, filtering, and concentrating under reduced pressure to specific gravity of 1.10-1.20 g/cm at 80 deg.C³Cooling the clear paste to room temperature, adding ethanol, controlling the volume concentration of the ethanol to be 70%, standing for 24h, filtering, recovering the ethanol, and concentrating to the specific gravity of 1.30-1.40 g/cm at 80 DEG C³The thick paste of (4);

(3) adding the glabrous greenbrier rhizome powder prepared in the step (1) into the thick paste prepared in the step (2), and uniformly mixing to obtain the effective component.

After the active ingredients are prepared by the method, tablets containing 1-5 g of crude drugs can be prepared by the method disclosed in the patent application example 1 with the publication number of CN105233150A, and the taking method of the tablets is as follows: the preparation is administered 3 times a day (once in the morning, noon and evening) for 5 tablets each time by an adult, and 7 days is a treatment course.

The medicine for preventing and treating myocardial ischemia-reperfusion injury can improve the cardiac function of a model rat, inhibit inflammatory reaction, regulate immunity, relieve the pathological change of myocardial fibrosis and relieve myocardial ischemia-reperfusion injury.

Drawings

FIG. 1 is a graph of the effect of psoriatin tablets after 30 days treatment on each group of rats, wherein (a) is an electrocardiogram of the rats, (b) is an ultrasound image, (c) is a histogram of left ventricular ejection fraction results, and (d) is a histogram of fractional shortening results; (c) in graphs (d), P < 0.01.

FIG. 2 is a bar graph of the effect of psoriatin tablets on serum levels in groups of rats, wherein (a) is creatine kinase isoenzyme (CK-MB), (b) is cardiac troponin I (CTn-I), and (c) is Lactate Dehydrogenase (LDH); in the figure, P < 0.05P < 0.01.

FIG. 3 is a bar graph showing the relative expression levels of inflammatory factors at myocardial injury sites in rats of various groups, wherein (a) is IL-4, (b) is IL-6, (c) is Foxp3, and (d) is IL-17; in the figure, P < 0.05P < 0.01.

FIG. 4 is a staining pattern of myocardial tissue of rats in each group, wherein (a) is an HE staining pattern and (b) is a Masson staining pattern.

Detailed Description

The present invention will be described in further detail with reference to the following examples and the accompanying drawings.

A. Experimental drugs (hereinafter called psoriasis tablet)

1. The raw material medicines and the proportion are as follows: 10 parts of curcuma zedoary, 20 parts of glabrous sarcandra herb, 20 parts of sinkiang arnebia root, 15 parts of red paeony root, 20 parts of glabrous greenbrier rhizome, 20 parts of dark plum fruit, 10 parts of liquoric root, 10 parts of Chinese angelica, 10 parts of szechuan lovage rhizome and 20 parts of rehmannia root.

2. The preparation method comprises the following steps:

(1) weighing rhizoma Smilacis Glabrae, pulverizing, and sieving with 80 mesh sieve to obtain rhizoma Smilacis Glabrae powder;

(2) weighing dried decoction pieces of rhizoma Curcumae, herba Pileae Scriptae, radix Arnebiae, radix Paeoniae Rubra, fructus mume, radix Glycyrrhizae, radix Angelicae sinensis, rhizoma Chuanxiong, and radix rehmanniae, decocting for three times, adding 11 times of water for the first time, and extracting for 2 hr; adding 9 times of water for the second and third times, and extracting for 1 hr; mixing extractive solutions, filtering, and concentrating under reduced pressure to specific gravity of 1.15g/cm at 80 deg.C³Cooling the fluid extract to room temperature, adding ethanol while controlling the volume concentration of ethanol to be 70%, standing for 24h, filtering, recovering ethanol, concentrating to specific gravity of 1.35g/cm at 80 deg.C³The thick paste of (4);

(3) mixing lactose, starch, dextrin and the soft extract obtained in step (2), drying, pulverizing, sieving with 80 mesh sieve, adding magnesium stearate and the rhizoma Smilacis Glabrae powder obtained in step (1), mixing, and making into tablet according to conventional method; wherein the adding amount of the lactose, the starch and the dextrin is 32.67 percent of the weight of the thick paste, the weight ratio of the lactose, the starch and the dextrin is 1:2:6, and the adding amount of the magnesium stearate is 0.67 percent of the weight of the thick paste.

B. Experiment of drug effect

Experiment of psoriasis tablet on myocardial function protection and inflammation inhibition of MIRI rats and myocardial fibrosis alleviation

1 materials and methods

1.1 Experimental materials

1.1.1 Experimental animals

50 male SPF SD rats with weight of 180-. Rat feeding conditions: the room temperature is 24 +/-1 ℃, the humidity is 40-80%, the illumination is 12h, the light and the shade are alternated, and the water and the food can be freely drunk and eaten. Adaptive feeding for 3-7 days before experiment. The experimental process complies with the relevant guidelines for the protection of experimental animals.

1.1.2 Main Instrument

Small animal ultra-high resolution B-ultrasound Vevo2100(FUJIFILM visual sonic Inc, Canada); 16-channel high-speed electrophysiological recording instrument (Millar, stz-0007593); small animal ventilator (HARVARD APPATUS CAT:55-7058, USA) microtome LEICA RM 2235; fully automated inverted fluorescence microscopy analysis system (NiKon T300); a centrifuge; a low temperature refrigerator (Sanyo, Japan), an ultra-low temperature refrigerator (Mitsubishi, China), a homogenizer (BD, USA), a desktop high speed refrigerated centrifuge (Eppendorf Co., Ltd., Germany), a PCR instrument, an electrophoresis apparatus (Bio-Rad Co., Ltd., USA), a 7500 sequence fluorescence quantitative PCR instrument (applied Biosystems, USA), and a PM-30 inverted microscope-computer image analysis system (Olympus, Japan)

1.1.3 reagents

Promega Eastep Super total RNA extraction kit; RNA reverse transcription kit (Thermo Fisher Scientific LOT 00662404); primer synthesis (huada gene); SYBR Green Super Mix (Bio-rad); hematoxylin; eosin (chemical reagent factory, guangzhou, china); masson staining kit (Mixin MST-8003/8004), CK-MB kit (constructed by Nanjing), CTn-I kit (constructed by Nanjing), and LDH kit (constructed by Nanjing).

1.1.4 preparation of liquid medicine

According to the regulation of 'traditional Chinese medicine pharmacological experimental methodology', the administration dosage of a rat is calculated according to a direct conversion formula of equivalent dosage between animals of different species, 7.5g (5 tablets multiplied by 3 times multiplied by 0.5g) of psoriasis tablets are taken by adults every day, the administration dosage of the rat is 6 times of that of the adults, the psoriasis tablets are taken and added with distilled water to be fully suspended to prepare a suspension containing 0.075g tablet weight per milliliter, and the suspension is stored at 4 ℃ for standby.

1.2 Experimental methods

1.2.1 preparation of rat MIRI model

All rats were fed routinely under the same conditions and fasted for 12h prior to surgery. The experimental animals are anesthetized by intraperitoneal injection with 1% sodium pentobarbital solution (40mg/Kg), after full anesthesia, the animals are inserted into an air tube and connected with a small animal respirator (harvard ventilator CAT:55-7058) to carry out mechanical ventilation, the tidal volume is 1ml/100g, the ventilation frequency is 70 times/minute, and the breathing ratio is 1: 1. needle electrodes were subcutaneously inserted through the right forelimb, left hind limb and right hind limb and connected to a multichannel electrophysiology apparatus (Millar, stz-0007593) to perform II lead monitoring. Fixing the mouse on an anatomical table, after preparing skin on the chest conventionally, carefully cutting the skin by about 1.5cm by using an ophthalmological scissors between 3 rd to 4 th ribs, separating the pectoralis major and the anterior serratus muscle bluntly, separating two sides by using a spreader for fixing, gently pushing the left lung downwards or outwards by using a physiological saline cotton ball, tearing the pericardium bluntly to expose the left anterior wall and the auricle of the heart, inserting a needle from the middle point of the lower edge of the left auricle by using a No. 7-0 nylon wire with a round needle, wherein the depth is about 1mm, withdrawing the needle between the right edge of the left auricle and the pulmonary artery cone, and ligating the anterior-inferior branch of the left coronary artery (about 2mm below the left auricle). Subsequently, both ends of the suture were passed through a polyethylene tube of 2mm diameter and 1.5cm in length to form a noose. Ligation was performed for 1 hour, and the ligation was released to allow reperfusion. And (5) performing layer-by-layer suture after 30min of reperfusion observation, and pulling out the ventilation tube when the rat generates autonomous swallowing reflex. The intraperitoneal injection of penicillin is 10 ten thousand U/100g after the operation. And keeping the temperature and waiting for reviving. The sham operation group only needs to thread the needle without pricking, and the rest treatment is the same as the experimental group. After revival, the animals in each group are fed regularly under the same environment.

Criteria for successful establishment of ischemia-reperfusion injury model

The successful implementation of myocardial ischemia treatment through ligation is characterized in that cyanosis or grayish white epicardium at the distal end of the ligature, synchronous II lead ECG shows that ST segment is obviously raised, and ventricular arrhythmia appears during ischemia; following restoration of myocardial blood perfusion by loosening the blocked LAD, the elevated ST segment is relatively reduced (> 50%) and the epicardial color of the ischemic area returns to ruddy. According to this criterion, the sham-operated group of rats finally included in the experiment totaled 13 rats, and the ischemia-reperfusion injury model of rats 26.

1.2.2 grouping and administration

Rats were randomly divided into 3 groups, i.e., a sham operation group (13), a model group (13), and a psoriasis treatment group (13) according to a random number table method. The administration by gavage is started on the 5 th day after the model building, 1 time per day, the administration by gavage tablet (0.75g tablet weight/kg) is carried out on the psoriasis tablet treatment group, the administration by distilled water with equal dose is carried out on the model group and the sham operation group, and the administration by gavage is continuously carried out for 30 days. The dosage is obtained by conversion according to the dosage of human body.

1.3 rat cardiac ultrasound, electrocardiographic testing

Echocardiography of left ventricular function

Weighing animals, carrying out isoflurane air anesthesia, clipping the left chest hair of a rat, dipping a cotton ball in a depilatory, smearing, cleaning with warm water after depilatory minutes, and then wiping with gauze; the limbs are fixed on the operation table. The left lateral decubitus is coated with a small amount of coupling agent on the front of the left chest, the high-frequency linear array probe is placed on the front of the left chest and pointed to the right, and the standard combination of the short axis section of the papillary muscle of the left ventricle and the long axis section of the left ventricle and Doppler ultrasound are selected for examination. The data measured for each cardiac cycle are averaged. Due to the fast heart rate of the murine animals, the profile and doppler blood flow spectrum scanning speed was set to >100 mm/s. And (3) observation indexes are as follows: left ventricular end-diastolic diameter (LVIDd), left ventricular end-systolic diameter, (LVIDs) ventricular interval end-diastolic thickness (IVSTd), ventricular interval end-systolic thickness (IVSTs), left ventricular posterior wall end-diastolic thickness (LVPWs), left ventricular posterior wall end-systolic thickness (LVPWs); left ventricular ejection fraction (EF%) and left ventricular minor axis shortening (FS%).

1.4 obtaining materials and preparing specimens

After 3% sodium pentobarbital (1.5ml/kg) is used for intraperitoneal injection and anesthesia, 3 ml/rat blood is taken from the inferior vena cava and used for an enzyme-linked immunosorbent assay of rat serum. Then randomly selecting 5 in each group, carrying out transcental perfusion, taking cardiac tissues for paraffin section staining, and calculating the myocardial fibrosis success of the cardiac tissues; after 8 random anesthetized animals in each group, the damaged part of the heart was removed on ice and placed in an EP tube, and then the heart was quickly frozen with liquid nitrogen and stored in a refrigerator at-80 ℃ for detecting the mRNA expression of the myocardial tissue.

1.5 rat serum enzyme-linked immunosorbent assay (ELISA)

The serum: naturally coagulating the blood at room temperature for 10-20min at 3000 rpm at 2000-.

Serum was diluted 5-fold for all experiments;

placing the kit at room temperature for balancing for half an hour before use;

respectively adding standard sample wells, zero wells and sample wells, and then adding 50 mu l of HPR reagent;

fourthly, lightly shaking, covering a sealing plate membrane, and incubating for 60min in an incubator at 37 ℃;

washing: carefully uncovering the sealing plate film, discarding the liquid, spin-drying, filling the cleaning solution into each hole with a row gun, standing for 30 seconds

Then discarding, repeating for 5 times, and patting dry;

sixthly, color development: adding 50 μ l of color-developing agent A into each well, adding 50 μ l of color-developing agent B, shaking gently, mixing, and developing at 37 deg.C in dark for 10 min;

and seventhly, terminating: stop solution 50. mu.l was added to each well to stop the reaction (blue color turned to yellow color immediately);

measurement of the formula (b): the absorbance (OD value) of each well was measured sequentially at a wavelength of 450nm with the blank well being zeroed. The determination should be adding

Stopping within 10 min;

ninthly, calculating: and calculating a regression equation of the standard curve according to the concentration and the OD value, and fitting the simulation curve to select a logistic curve.

1.6 Total RNA extraction and isolation of myocardial tissue (Promega Eastep Super)

50mg of tissues are transferred into a mortar precooled by liquid nitrogen and baked for 8 hours in an oven at 180 ℃, a specimen is ground into powder, and the liquid nitrogen is continuously added in the grinding process; adding a proper amount of RNA lysate and diluent and grinding; mixing with pipette, standing at room temperature for 3-5 min or heating at 70 deg.C for 3 min to increase RNA yield.

② after mixing, 12000-14000Xg for 5 minutes. The supernatant was carefully aspirated.

③ adding 0.5 time of absolute ethyl alcohol of the volume of the supernatant, and mixing evenly.

Fourthly, the mixed solution is transferred to a centrifugal column, and is centrifuged for 1 minute by 12000-14000Xg, and the filtrate is discarded. Add 600 u l RNA washing solution washing, 12000 and 14000Xg centrifugal 45 seconds, discard filtrate.

Fifthly, adding 50 mul of DNase I incubation liquid to incubate for 15 minutes.

Sixthly, 600 mu l of RNA washing solution is added, 12000-14000Xg is centrifugated for 45 seconds, and the filtrate is discarded. Repeating twice

Seventhly, resetting the centrifugal column on the collecting pipe, and centrifuging for 2 minutes at 12000-14000 Xg.

Will spin the column on the elution tube, add 50 u l of nuclease-free water, 12000-.

1.7 reverse transcription

The synthesis of cDNA was performed according to the procedures in the kit instructions. A10. mu.l sample reaction containing 2. mu.g of gRNA and 2. mu.l of random primer was added with nuclease-free water to 3.2. mu.l, and heated at 70 ℃ for 5 minutes. Immediately frozen on ice for at least 5 minutes, add 4.8. mu.l of reverse transcription mix and flash centrifuge for 10 seconds. Mixing, annealing, heating to 25 deg.C, incubating for 5 min, incubating at 37 deg.C for 120 min, heating to 85 deg.C for 5 min, cooling to 4 deg.C, taking out, and storing at-20 deg.C. The reverse transcription mixture is shown in table 1 below:

table 1: 10 mul reagent system for reverse transcription of RNA into cDNA

1.8Real Time-PCR analysis

The reaction system contained in 20. mu.l of a real-time reaction system: mu.l of univarial SYBR Green supermix (2X), 5. mu.l of the upstream and downstream primer mix (each at a final concentration of 500nM), and 5. mu.l of 20-fold diluted cDNA.

Covering 96-well plate with special fluorescent quantitative sealing film, instantaneous centrifuging, and placing into fluorescent quantitative PCR instrument for amplification for 40 cycles at 95 deg.C 30s, 60 deg.C 30s, and 72 deg.C 90s for denaturation, annealing, and extension.

GAPDH was used as a reference gene and the assay was repeated 3 times per sample. The relative amount of mRNA is expressed as Ct value, and the average relative expression is analyzed by the 2-Delta Ct calculation method.

The primer sequences used are shown in table 2 below:

table 2: primer sequences

1.9HE staining

After 30 days of dosing, the rats were hearted, fixed, sectioned, and HE stained as follows.

Hematoxylin-eosin (HE) staining: slicing paraffin samples with a thickness of 4.5-5 μm with a paraffin microtome, continuously slicing, and staining with baking sheet for 20 minutes, xylene for 2 × 10 minutes, absolute ethanol for 2 × 2 minutes, 95% ethanol for 1 minute, 80% ethanol for 1 minute, 70% ethanol for 1 minute, water washing for 1 minute, hematoxylin for 8 minutes, hematoxylin for 10 minutes, water washing for 2 × 1 minute, 0.5% hydrochloric acid ethanol for 10 seconds, water washing for 10 minutes, eosin for 2 minutes, water washing for 1 minute, 80% ethanol for 5 seconds, 85% ethanol for 5 seconds, 90% ethanol for 5 seconds, 95% ethanol for 1 minute, absolute ethanol for 2 × 2 minutes, absolute ethanol for 3 minutes, xylene for 2 × 2 minutes, and sealing with neutral gum after staining.

2.0Masson staining

According to the kit (maixin MST-8003/8004) instructions:

(1) the tissue paraffin blocks are sliced, each slice is 4.5 mu m thick, the slices are dried by a 65 ℃ slice drying instrument, and the slices are dried in a 65 ℃ electric heating constant temperature air blast drying oven overnight.

(2) 1 drop of Masson's complex stain (reagent A) was added to the tissue and stained for 5 minutes. The dye liquor was rinsed off with distilled water.

(3) Phosphomolybdic acid (reagent C) staining 1 drop was added to the tissue, followed by staining for 5 minutes, and then the stain was spun dry.

(4) 1 drop (100. mu.l) of aniline blue (reagent D) was added directly dropwise and stained for 5 minutes with distilled water and rinsed slightly.

(5) Dropping 1 drop of differentiation solution (reagent B) for differentiation for 30-60 seconds (2 times)

(6) Dehydrating with 95% ethanol and anhydrous ethanol, and sealing.

2.1 statistical treatment

Statistical analysis SPSS20.0 statistical analysis software was used. The measured data conforms to the normal distribution and adopts mean plus or minus standard deviation

The measured data comparison among the groups is in accordance with the conditions, the analysis of variance is designed by adopting a single factor, the comparison between every two groups adopts an LSD method, and the difference is more than 0.05, so that the statistical significance is achieved.

Second, results and analysis

1. The effect of psoriasis tablets on the cardiac function of rats in each group;

electrocardiogram: compared with the MIRI model group, the psoriasis tablet treatment group has less ST segment shift and improved myocardial ischemia symptoms, and the results are shown in FIG. 1 (a).

Cardiac ultrasound: the psoriasis tablet can obviously improve the myocardial contraction function and compliance and protect the myocardium from further damage, and the result is shown in figure 1 (b).

The LVEF value of the MIRI group is reduced to 43.09 +/-2.117 (%) on the 35 th day after operation, the LVFS value is 24.86 +/-1.461, the LVEF value of the psoriasis tablet is recovered to 67 +/-1.728 (%), and the LVFS value is 38.95 +/-1.193; it is indicated that the psoriasis treating tablet can restore cardiac muscle function effectively.

Comparisons between groups were made due to variance differences, and examined by Dunnett's T3 test. The results show that after 30 days, the left ventricular ejection fraction and the shortened fraction of the model group have P <0.01 compared with the sham operation group, which indicates that the model is stable; the left ventricular ejection fraction and the foreshortening fraction of the model group compared to the treatment group, P <0.01, indicated that the drug treatment was effective, and the results are shown in fig. 1(c) and fig. 1 (d).

2. The influence of the psoriasis tablet on the serum CK-MB, CTn-I and LDH of each group of rats;

myocardial damage caused damage to the biological membrane system, creatine kinase isoenzyme (CK-MB), cardiac troponin I (CTn-I) and Lactate Dehydrogenase (LDH) entering the blood from within the damaged myocardium, creatine kinase isoenzyme (CK-MB), cardiac troponin I (CTn-I) and Lactate Dehydrogenase (LDH) levels in the psoriasis patch treatment group were much lower than in the MIRI model group, and statistical analysis was performed using independent sample t-test. The results showed that the difference was statistically significant, P <0.05, compared to the sham group. The psoriasis treating tablet can effectively improve myocardial damage and relieve the myocardial infarction process.

3. The effect of the psoriasis treating tablet on the relative expression quantity of the inflammatory factors at the myocardial damaged parts of the rats in each group is as follows:

IL-4 is capable of stimulating B cell proliferation and inhibiting IFN-gamma production. And (3) carrying out statistical analysis by adopting an independent sample t test, and displaying the result: the relative expression of IL-4 gene in the treated group was higher than that in the model group, and the difference was statistically significant (P < 0.05), and the results are shown in FIG. 3 a.

IL-6 can promote the secretion of inflammatory cytokines and aggravate inflammatory reaction by stimulating the differentiation of T cells, and can promote the generation of autoantibodies and induce autoimmune lesions on the other hand. Meanwhile, IL-6 is an important proinflammatory factor, and the concentration of the IL-6 in the myocardial ischemia-reperfusion injury process reflects the degree of myocardial injury. And (3) carrying out statistical analysis by adopting an independent sample t test, and displaying the result: the relative expression of IL-6 gene in the model group was higher than that in the treatment group, and the difference was statistically significant (P < 0.05), and the results are shown in FIG. 3 b.

The Foxp3 mutation can cause autoimmune endocrine diseases, inflammatory bowel diseases, fatal infections, severe allergic reactions and the like, and research proves that the Foxp3 plays an important role in immune regulation. And (3) carrying out statistical analysis by adopting an independent sample t test, and displaying the result: the relative expression of the Foxp3 gene in the model group was much higher than that in the treatment group, and the difference was significant (P < 0.01), and the result is shown in FIG. 3 c.

The IL-17 family also has a wide variety of roles, including recruitment of macrophages, neutrophils to inflammatory tissue, action on endothelial cells and macrophages, production of IL-6, IL-1, TNF- α, C-Reactive protein (CRP) and metallo-matrix protease mediated inflammatory responses, increased inflammatory cell adhesion function by modulating chemokine expression, and promotion of apoptosis. Studies have shown cytokine and inflammatory mediator release from damaged tissues as: IL-6, TNF alpha, IL-10, IL-17, etc. can cause apoptosis and distant organ damage. Inhibiting IL-17 can delay inflammation and relieve myocardial pathological injury. And (3) carrying out statistical analysis by adopting an independent sample t test, and displaying the result: the relative expression level of IL-17 gene in the model group was much higher than that in the treatment group, and the difference was statistically significant (P < 0.05), and the results are shown in FIG. 3 d.

4. Effect of psoriatic tablets on myocardial fibrosis in MIRI rats:

myocardial HE staining results: the arrangement of the myocardium of the rats in the sham operation group is approximately regular, and the form of the myocardium is relatively complete; the rats in the model group had disorganized myocardial arrangement, myocardial rupture, and increased area of myocardial fibrosis accompanied by more inflammatory cell infiltration. The psoriasis tablet rat myocardial tissue fiber arrangement is less regular, myocardial fibrosis area is smaller, and inflammatory cell number is less. The results are shown in FIG. 4 a. Results of myocardial Masson staining: masson staining showed large areas of MIRI staining by aniline blue, indicating massive deposition of collagen, fibronectin and other substrates. In the psoriasis treating group, the above lesions are obviously improved. The results are shown in FIG. 4 b. Calculating the myocardial fibrosis area ratio: after Masson staining of R myocardial sections from each group of rats, 5 fields were randomly selected on the sections, cell size counting was performed using Image-pro software and collagen volume fraction (CVF%) was calculated. CVF% ═ area of myocardial collagen/total area of myocardial cells 100%. Performing normalization test and homogeneity test of variance on the data to obtain normalized distribution, and performing statistical analysis by Wilcoxon test in non-parametric test method

A description will be given. The results show that the P is less than 0.01 and the difference is more obvious when the treatment group is compared with the model group.

Table 3: effect of myocardial fibrosis in rats of Each group

Note: comparing model group with sham operation group, P is less than 0.01; psoriasis tablet group compared with model group by P < 0.01.

Sequence listing

<110> southern medical university

Application of psoriasis treating traditional Chinese medicine in preparation of medicine for preventing and treating myocardial ischemia reperfusion injury

<160> 10

<170> SIPOSequenceListing 1.0

<210> 1

<211> 24

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 1

cggagtcaac ggatttggtc gtat 24

<210> 2

<211> 24

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 2

agccttctcc atggtggtga agac 24

<210> 3

<211> 24

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 3

tccatgcacc gagatgtttg tacc 24

<210> 4

<211> 24

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 4

gcaagtattt ccctcgtagg atgc 24

<210> 5

<211> 25

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 5

gccagagtca ttcagagcaa tactg 25

<210> 6

<211> 24

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 6

ttgggatatc aggtttctgg atgg 24

<210> 7

<211> 21

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 7

gcccaccagc atcttctcca c 21

<210> 8

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 8

ccatccatgt gcctgatgct 20

<210> 9

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 9

tgagctggct gcaattctgg 20

<210> 10

<211> 24

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 10

atctagctgc tctgcatgag gtga 24

Claims (2)

1. The application of the traditional Chinese medicine for treating psoriasis in preparing the medicine for preventing and treating myocardial ischemia-reperfusion injury is characterized in that the effective components of the traditional Chinese medicine for treating psoriasis comprise powder prepared from 20 parts by weight of glabrous greenbrier rhizome and aqueous extracts of the following raw material medicines:

10 parts of curcuma zedoary, 20 parts of glabrous sarcandra herb, 20 parts of lithospermum, 15 parts of red paeony root, 20 parts of dark plum fruit, 10 parts of liquorice, 10 parts of Chinese angelica, 10 parts of szechuan lovage rhizome and 20 parts of rehmannia root.

2. The use according to claim 1, wherein the active principle is prepared by the following method:

(1) pulverizing rhizoma Smilacis Glabrae, and sieving with 80 mesh sieve to obtain rhizoma Smilacis Glabrae powder;

(2) taking rhizoma zedoariae, glabrous sarcandra herb, lithospermum, red paeony root, dark plum fruit, liquorice root, Chinese angelica, Szechuan lovage rhizome and rehmannia root decoction pieces, decocting and extracting for three times by adding 10-12 times of water for the first time, decocting and extracting for 2 hours, and respectively adding 8-10 times of water for the second time and the third time, and decocting and extracting for 1 hour; mixing the extractive solutions, filtering, and concentrating under reduced pressure until the specific gravity of the extract measured by heat is 1.10-1.20 g/cm at 80 deg.C³Cooling the clear paste to room temperature, adding ethanol, controlling the volume concentration of the ethanol to be 70%, standing for 24h, filtering, recovering the ethanol, and concentrating to the specific gravity of 1.30-1.40 g/cm at 80 DEG C³The thick paste of (4);

(3) adding the glabrous greenbrier rhizome powder prepared in the step (1) into the thick paste prepared in the step (2), and uniformly mixing to obtain the effective component.

Images