Application of scopalactone in treatment of pulmonary arterial hypertension

Abstract

The invention discloses application of scopalactone in treatment of pulmonary arterial hypertension, and relates to the technical field of medicinal chemistry. The scoparone is applied to preparation of the medicine for treating pulmonary arterial hypertension. The medicine comprises scopalactone, a pharmaceutically acceptable carrier, a diluent and an excipient, the medicine further comprises other medicines for treating the pulmonary arterial hypertension, and the other medicines for treating the pulmonary arterial hypertension are selected from a prostacyclin medicine, an endothelin receptor antagonist and a phosphodiesterase-5 inhibitor. The scopalactone is applied to preparation of the medicine for treating pulmonary arterial hypertension, right heart insufficiency caused by hypoxia can be remarkably reversed, and the right heart function is effectively improved by reducing the transverse diameter of the right ventricle and the transverse diameter of the right atrium and improving the indexes such as tricuspid valve ring plane contraction displacement; the average pulmonary arterial pressure and right ventricular systolic pressure can be obviously reduced, and hemodynamic parameters are improved; the area and thickness of a blood vessel wall can be remarkably reduced, the blood vessel structure is improved, and pulmonary arteriole reconstruction is relieved.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of pharmaceutical chemistry, and particularly relates to application of scoparone in treatment of pulmonary arterial hypertension. BACKGROUND

[0002] Pulmonary arterial hypertension (PAH) is a severe cardiovascular disease characterized by elevated pulmonary arterial pressure, leading to excessive right heart load and eventually causing right heart failure. Currently, the drugs used for treating pulmonary arterial hypertension in clinical practice mainly include prostacyclin drugs, endothelin receptor antagonists and phosphodiesterase-5 inhibitors, etc. However, these drugs all have certain side effects and limitations. For example, prostacyclin drugs can cause adverse reactions such as headache and diarrhea; endothelin receptor antagonists can cause abnormal liver function; and phosphodiesterase-5 inhibitors can cause side effects such as hypotension. Therefore, it is of great significance to find new effective therapeutic drugs.

[0003] In recent years, natural products have shown great potential in the treatment of cardiovascular diseases. Scoparone (Scop), as an effective component extracted from traditional Chinese medicine, has multiple biological activities such as antioxidant, anti-inflammatory and anti-fibrosis. Studies have shown that Scop has good therapeutic effect in various cardiovascular disease models, but its application in the treatment of pulmonary arterial hypertension has not been fully explored. Based on this, the present application aims to explore the potential application value of Scop in the treatment of pulmonary arterial hypertension, and to systematically evaluate the effects of Scop on right heart function, hemodynamic parameters and pulmonary arteriole remodeling by constructing a hypoxia-induced rat model of pulmonary arterial hypertension, so as to provide a scientific basis for its clinical application. SUMMARY

[0004] The present application aims to solve the problems existing in the prior art and provides application of scoparone in treatment of pulmonary arterial hypertension.

[0005] To achieve the above-mentioned purpose, the present application adopts the following technical solutions:

[0006] The application of scoparone in treatment of pulmonary arterial hypertension applies scoparone in the preparation of a drug for treating pulmonary arterial hypertension.

[0007] Preferably, the drug comprises scoparone and a pharmaceutically acceptable carrier, diluent or excipient.

[0008] Preferably, the drug further comprises other drugs for treating pulmonary arterial hypertension, and the other drugs for treating pulmonary arterial hypertension are selected from prostacyclin drugs, endothelin receptor antagonists and phosphodiesterase-5 inhibitors.

[0009] The application of a kind of scabrosin in treating pulmonary arterial hypertension, the scabrosin in treating pulmonary arterial hypertension, the treatment effect is verified by experiment, the treatment dosage is determined, and specific steps include as follows:

[0010] S1: hypoxia-induced rat pulmonary arterial hypertension model;

[0011] S2: right heart function detection by echocardiography;

[0012] S3: right heart catheter measurement of hemodynamic parameters;

[0013] S4: HE staining observation of pulmonary arterioles;

[0014] S5: result analysis.

[0015] Preferably: in the S1, the hypoxia-induced rat pulmonary arterial hypertension model is established, and the specific steps are as follows:

[0016] S11: male specific pathogen-free level SD rats are bred in an independent cage feeding system, and free feeding is carried out;

[0017] S12: the rats are randomly divided into a control group, a Scop group, a hypoxia group and a hypoxia+Scop group;

[0018] S13: the rats in the hypoxia group and the hypoxia+Scop group are subcutaneously injected with SU5416 and bred in a normal pressure hypoxia incubator, and the other groups are injected with an equal volume of DMSO / PEG300 / Tween-80 / normal saline mixture and bred in a normal pressure and normal oxygen condition;

[0019] S14: after 7 days, the rats in the Scop group and the hypoxia+Scop group are fed with 50mg / kg Scop, and 1mg / kg Scop is fed every other day to maintain the drug concentration;

[0020] S15: 21 days after Scop treatment, the rats are lightly anesthetized with isoflurane, the right heart function is evaluated by echocardiography, and the hemodynamic parameters are measured by right heart catheter.

[0021] Preferably: in the S13, the rats in the hypoxia group and the hypoxia+Scop group are subcutaneously injected with SU5416, the dosage is 20mg / kg, once a week, and the O2 concentration in the normal pressure hypoxia incubator is controlled at 10%; the other groups are injected with an equal volume of DMSO / PEG300 / Tween-80 / normal saline mixture, and the proportion of each group is controlled at 2:8:1:9.

[0022] Preferably: in the S2, the echocardiography is detected by using a flyinuo VINN06 LAB rat ultrasonic imaging system, and the specific steps are as follows:

[0023] S21: After the rats were anesthetized, they were fixed on the 37℃ animal operating table in a supine position, and the chest hair was removed with depilatory cream;

[0024] S22: The probe was placed at the root of the parasternal aorta to perform short-axis imaging, and the right ventricular transverse diameter, right atrial transverse diameter, pulmonary artery diameter, and aortic diameter were measured;

[0025] S23: When the apical four-chamber section was used, M-mode ultrasound was used to measure the tricuspid annular plane displacement in the systolic phase;

[0026] S24: The probe was placed at the root of the parasternal aorta to perform short-axis imaging at the pulmonary valve orifice, and the pulmonary valve blood flow spectrum, acceleration time, ejection time, and pulmonary valve regurgitation spectrum were measured using spectral Doppler mode.

[0027] Preferably, in S3, the specific steps are:

[0028] S31: After the rats were anesthetized, they were fixed on the 37℃ animal operating table in a supine position, and the right neck was incised to expose the right jugular vein;

[0029] S32: At the 1 / 3 of the jugular vein, the blood vessel was cut 1 / 3 with ophthalmic scissors at a 45° angle towards the ventricle to form a V-shaped incision of 2-3mm;

[0030] S33: A polyethylene catheter filled with 0.3% heparin saline was inserted into the right atrium through the port, and an MP150 multi-channel physiological recorder was used to record the pressure at each site;

[0031] S34: The direction was kept unchanged, the catheter continued to advance and rotate counterclockwise to hook the tricuspid valve septum with the catheter head pointing to the left, enter the right ventricle, observe the waveform changes, and further advance the catheter to the pulmonary artery to record the pulmonary artery pressure.

[0032] Preferably, in S4, the pulmonary arteriole remodeling is observed by HE staining, and the specific steps are as follows:

[0033] S41: The heart and lung were flushed with physiological saline, the lung tissue was cut and rinsed with PBS to remove blood;

[0034] S42: The lung tissue was immersed in 4% paraformaldehyde and stored at 4℃ for 48 hours;

[0035] S43: Tissue wax blocks were prepared for embedding and sectioning for HE staining;

[0036] S44: The staining was observed under an electron microscope and photographed, and 5-8 pulmonary arterioles with a diameter <100mm were randomly selected from each section;

[0037] S45: The percentage of the medial muscle layer of the pulmonary arteriole to its outer diameter and the percentage of the total area of the blood vessel were calculated.

[0038] Preferably: in the S45, the specific mode is to measure the vascular peripheral length and the lumen peripheral length by an Image Pro Plus 6.0 color image analysis system, to calculate the vascular outer diameter, the lumen inner diameter, the total vascular area and the lumen area, and finally to calculate the percentage of the pulmonary arteriole media muscle layer to the outer diameter and the percentage of the pulmonary arteriole media muscle layer to the total vascular area.

[0039] The beneficial effects of the present application are:

[0040] 1. The present application applies scabrosin to the preparation of drugs for treating pulmonary arterial hypertension, which can significantly reverse right heart dysfunction caused by hypoxia, effectively improve right heart function by reducing the right ventricular transverse diameter, the right atrial transverse diameter and the tricuspid annular plane systolic displacement, etc., can significantly reduce the mean pulmonary arterial pressure and the right ventricular systolic pressure, thereby improving the hemodynamic parameters, and can significantly reduce the vascular wall area and thickness, improve the vascular structure, and thus alleviate the pulmonary arteriole remodeling.

[0041] 2. The present application comprehensively evaluates the therapeutic effect of Scop on the pulmonary arterial hypertension rat model through echocardiography, right heart catheter measurement and HE staining, etc., to ensure the scientificity and reliability of the results; and adopts random grouping, control experiments and various detection means to ensure the objectivity and accuracy of the experimental results. BRIEF DESCRIPTION OF DRAWINGS

[0042] Figure 1 Fig. 1 is a representative echocardiogram of each group of rats in the experiment of the present application;

[0043] Figure 2 Fig. 2 is a statistical analysis diagram of the PAT / PET(A), PAT(B), TAPSE(C), RATD(D), PA / AO(E) and RVTD(F) indexes of the right heart function in the experiment of the present application;

[0044] Figure 3 Fig. 3 is a representative image of the RVSP and mPAP in the experiment of the present application and a statistical analysis diagram of the RVSP and mPAP;

[0045] Figure 4 Fig. 4 is a representative image of the pulmonary arteriole of each group of rats in the hypoxia-induced pulmonary arterial hypertension model in the experiment of the present application and a statistical analysis diagram of the vascular wall thickness, the vascular diameter, the vascular wall area and the total vascular area. DETAILED DESCRIPTION

[0046] The technical solutions of the present application will be further described in detail in combination with the specific embodiments.

[0047] Example 1:

[0048] The application of scoparone in treating pulmonary arterial hypertension, the treatment effect is verified through experiments, the treatment dosage is determined, and the specific steps include the following steps:

[0049] S1: hypoxia-induced pulmonary arterial hypertension model in rats;

[0050] S2: cardiac ultrasound detection of right heart function;

[0051] S3: right heart catheter measurement of hemodynamic parameters;

[0052] S4: HE staining observation of pulmonary arterioles;

[0053] S5: result analysis.

[0054] I. Experimental materials

[0055] Scoparone (Scop) (BP1272) was purchased from Chengdu Purify Science and Technology Development Co., Ltd. (www.biopurify.cn). SU5416 (A3847) was purchased from APExBIO. Other reagents were purchased from ordinary reagent manufacturers. The reagents used are of analytical purity.

[0056] II. Experimental method

[0057] 1. Hypoxia-induced pulmonary arterial hypertension model in rats

[0058] Male specific pathogen-free grade SD rats (weight 180-220g) were purchased from the Experimental Animal Center of Chongqing Medical University and were raised in the independent cage feeding system of the animal center, and were free to eat. The rats were randomly divided into Ctrl, Scop, hypoxia and hypoxia+Scop groups, and the rats in the hypoxia and hypoxia+Scop groups were subcutaneously injected with SU5416 (20mg / kg, once a week) and raised in a normal pressure hypoxia incubator (10% O2), and other rats were injected with an equal volume of DMSO / PEG300 / Tween-80 / normal saline (2:8:1:9) mixture and raised in normal pressure and normal oxygen conditions (21% O2). After 7 days, the Scop and hypoxia+Scop groups of rats were fed with 50mg / kg Scop. In order to maintain the drug concentration, 1mg / kg Scop was fed every other day. The control group and the hypoxia group of rats were fed with an equal volume of DMSO and PBS mixture. 21 days after Scop treatment, the rats were lightly anesthetized with isoflurane (2%), the right heart function was evaluated by echocardiography, and the hemodynamic parameters were determined by right heart catheter.

[0059] 2. Cardiac ultrasound detection of right heart function

[0060] The rat ultrasonic imaging system of Vingmed Ultrasound VINN06LAB was used for cardiac ultrasonic detection, and the specific operation was as follows: after the rats were anesthetized with 2% isoflurane, they were fixed on the 37°C animal operating table in a supine position, and the chest hair was removed with depilatory cream before ultrasonic detection. First, the probe was placed in the short axis section of the aortic root to image the right ventricular transverse diameter, right atrial transverse diameter, pulmonary artery diameter and aortic diameter; when the four-chamber apical section was used, M-mode ultrasonic measurement was used to measure the tricuspid annular plane displacement; finally, the probe was placed in the short axis section of the aortic root of the pulmonary valve to measure the pulmonary valve blood flow spectrum, acceleration time, ejection time and pulmonary valve regurgitation spectrum by using the spectrum Doppler mode.

[0061] 3. Right heart catheterization to measure hemodynamic parameters

[0062] After the rats were anesthetized with 2% isoflurane, they were fixed on the 37°C animal operating table in a supine position, the right jugular vein was exposed by cutting the skin on the right side of the neck. At 1 / 3 of the jugular vein, the blood vessel was cut 1 / 3 obliquely at 45° with ophthalmic scissors towards the ventricle to form a V-shaped incision of 2-3 mm. A polyethylene catheter (inner diameter 0.5 mm, outer diameter 0.9 mm) filled with 0.3% heparin saline was sent into the right atrium through the port, and an MP150 multi-channel physiological recorder (BIOPAC Systems, CA, United States) was used to record the pressure at each part. When the pressure sensor is in the atrium, we can see the right atrial waveform with small waves, gentle curves, and no obvious difference between diastolic pressure and systolic pressure. Keeping the direction unchanged, the catheter continues to advance and rotate counterclockwise to hook the tricuspid valve septum with the catheter head pointing to the left and down, and continue to advance to enter the right ventricle. The right ventricular waveform rises and falls suddenly, and the blood pressure at the trough is close to 0 mmHg. Further advancing the catheter and slightly counterclockwise rotation can reach the pulmonary artery, and the pulmonary artery waveform is observed. At this time, the catheter can be fixed, and after the waveform is stable, the pulmonary artery pressure can be recorded.

[0063] 4. HE staining to observe pulmonary arterioles

[0064] After the right heart catheterization, the heart and lung were perfused with normal saline, and the lung tissue was cut and washed with PBS to remove blood. The left upper lobe of the lung was immersed in 4% paraformaldehyde at 4°C for 48 h, and then sent to Hunan Aifang Biological Technology Co., Ltd. for embedding and sectioning (thickness 5 pm) for HE staining. Each slice was observed under an electron microscope (x200) and photographed, and 5-8 pulmonary arterioles with a diameter <100 mm were randomly selected from each slice. The ImageProPlus6.0 color image analysis system was used to measure the vessel perimeter and lumen perimeter, and then calculate the external diameter (ED), internal diameter (ID), total area (TA), and lumen area (LA). Finally, the percentage of the pulmonary arteriole media muscle layer to its external diameter (WT%) and the percentage of the wall area to the total vessel area (WA%) were calculated. WT% = (ED-ID) / ED x 100%; WA% = (TA-LA) / TA x 100%. The data were arranged and plotted using GraphPad Prism 9.

[0065] III. Results and picture display

[0066] 1. Scop can improve the right heart function of hypoxia-induced pulmonary hypertension rats

[0067] In the previous stage, we constructed a hypoxia-induced pulmonary hypertension rat model and detected the effect of Scop on the right heart function of pulmonary hypertension rats. As shown in FIGS. 1A-1F, Scop can reverse the right heart dysfunction caused by hypoxia. The key parameters PAT / PET, PAT, and TAPSE of the right heart function of hypoxic rats were decreased, and Scop significantly reversed these parameters. The key parameters RATD, RVTD, and PA / AO reflecting the right heart structure remodeling were increased by 1.0-1.5 times by hypoxia, but Scop significantly reduced these parameters. Figure 1 Figure 2

[0068] Figures 1-2 ​​Scop can improve right heart function in MCT-induced pulmonary hypertension rats Wild-type male SD rats were randomly divided into Ctrl, Scop, hypoxia and hypoxia + Scop groups. The rats in the hypoxia and hypoxia + Scop groups were subcutaneously injected with SU5416 (20 mg / kg, once a week) and raised in a normobaric hypoxic chamber (10% O2). The other rats were injected with an equal volume of DMSO / PEG300 / Tween-80 / normal saline (2:8:1:9) mixture and raised in a normobaric normoxic condition (21% O2). After 7 days, the rats in the Scop and hypoxia + Scop groups were gavaged with 50 mg / kg Scop. To maintain the drug concentration, 1 mg / kg Scop was gavaged every other day. The control and hypoxia group rats were gavaged with an equal volume of DMSO and PBS mixture. Twenty-one days after Scop treatment, the rats were mildly anesthetized with isoflurane (2%), and right heart function was evaluated by echocardiography. Figure 1 : Representative images of echocardiography in each group; Figure 2 (A-F) Statistical analysis of right heart function-related indicators: PAT / PET (A), PAT (B), TAPSE (C), RATD (D), PA / AO (E) and RVTD (F). Data are expressed as mean ± SEM, n = 6. ***, p < 0.001; **, p < 0.01; *, p < 0.05. RVTD: right ventricular transverse diameter; RATD: right atrial transverse diameter; PA: pulmonary artery diameter; AO: aortic diameter; PET: pulmonary artery ejection time; PAT: pulmonary artery acceleration time; TAPSE: tricuspid annular plane systolic excursion.

[0069] 2. Scop can improve hemodynamic parameters in hypoxia-induced pulmonary hypertension rats

[0070] We observed the effect of Scop on hemodynamic parameters. As shown in A-C, the increased RVSP and mPAP induced by hypoxia were reversed by Scop. Figure 3

[0071] Figure 3 . Scop can improve hemodynamic parameters in pulmonary hypertension rats Representative images of hemodynamic parameters (A) RVSP and mPAP; (B) statistical analysis of RVSP; (C) statistical analysis of mPAP. Data are expressed as mean ± SEM, n = 6. **, p < 0.01; ***, p < 0.001. RVSP: right ventricular systolic pressure; mPAP: mean pulmonary arterial pressure.

[0072] 3. Scop reduces pulmonary arteriole remodeling in pulmonary hypertension rats

[0073] We detected the effect of Scop on pulmonary arteriole remodeling in a hypoxia-induced rat pulmonary hypertension model. As shown in A-C, the increased RVSP and mPAP induced by hypoxia were reversed by Scop.​Figure 4 As shown in FIGS. 12A-C, both hypoxia-induced increases in vascular wall area / total vessel area and vascular wall thickness / vessel diameter were reversed by Scop (p<0.001). The mean value of vascular wall area / total vessel area in the hypoxia group was 78.60±9.48%, while that in the hypoxia+Scop group was 48.53±5.09%. The mean value of vascular wall thickness / vessel diameter in the hypoxia group was 54.92±10.60%, while that in the hypoxia+Scop group was 28.34±3.54%.

[0074] Figure 4 Scop attenuates pulmonary arteriole remodeling in rats with pulmonary hypertension. Rats were sacrificed after evaluation of right heart function and hemodynamic parameters in the hypoxia-induced rat model of pulmonary hypertension. Fresh lung tissues were fixed with 4% paraformaldehyde and embedded in paraffin, and sections with a thickness of 5 μm were cut. The sections were stained with hematoxylin-eosin (HE) and photographed under a microscope. (A) Representative images of pulmonary arterioles in rats in each group in the hypoxia-induced rat model of pulmonary hypertension, scale bar = 50 μm; (B) Statistical analysis of vascular wall thickness / vessel diameter in rats in the hypoxia-induced rat model of pulmonary hypertension; (C) Statistical analysis of vascular wall area / total vessel area in rats in the hypoxia-induced rat model of pulmonary hypertension. Five different fields of each sample were selected for imaging for analysis. Data are expressed as mean ± SEM, n = 6. ***, p < 0.001. wall thickness / vessel diameter: vascular wall thickness / vessel diameter; wall area / total vessel area: vascular wall area / total vessel area.

[0075] The above merely describes preferred specific embodiments of the present application, but the protection scope of the present application is not limited thereto, and any person skilled in the art, according to the technical solution and inventive concept of the present application, can make equivalent replacements or changes within the technical scope disclosed by the present application, which should be encompassed within the protection scope of the present application.

Claims

1. Use of a costunolide in the treatment of pulmonary arterial hypertension, characterized in that, The scopenin is applied to the preparation of a drug for treating pulmonary arterial hypertension.

2. The use of a scullcaplactone according to claim 1 for the treatment of pulmonary arterial hypertension, characterized in that, The drug comprises scopenin and a pharmaceutically acceptable carrier, diluent, excipient.

3. The use of a scullcaplactone according to claim 1 for the treatment of pulmonary arterial hypertension, characterized in that, The drug also comprises other drugs for treating pulmonary arterial hypertension, and the other drugs for treating pulmonary arterial hypertension are selected from the group consisting of prostacyclin drugs, endothelin receptor antagonists and phosphodiesterase-5 inhibitors.

4. Use of a costunolide for the treatment of pulmonary arterial hypertension, characterized in that, The scopenin in the treatment of pulmonary arterial hypertension is verified by experiments, and the treatment amount is determined, and the specific steps include the following steps: S1: an anoxic-induced pulmonary arterial hypertension model of a rat is established; S2: right heart function is detected by echocardiography; S3: hemodynamic parameters are measured by a right heart catheter; S4: pulmonary arterioles are observed by HE staining; S5: results are analyzed.

5. The use of a scullcap-12 in the treatment of pulmonary arterial hypertension according to claim 4, characterized in that, In S1, the anoxic-induced pulmonary arterial hypertension model of the rat is established, and the specific steps are as follows: S11: male specific-pathogen-free grade SD rats are bred in an independent cage feeding system, and are free to eat; S12: the rats are randomly divided into a control group, a Scop group, an anoxic group and an anoxic+Scop group; S13: the rats in the anoxic group and the anoxic+Scop group are subcutaneously injected with SU5416 and bred in a normal-pressure hypoxia incubator, and the other groups are injected with an equal volume of a DMSO / PEG300 / Tween-80 / normal saline mixture and bred in a normal-pressure normoxia environment; S14: after 7 days, the rats in the Scop group and the anoxic+Scop group are fed with 50mg / kg Scop, and 1mg / kg Scop is fed every other day to maintain the drug concentration; S15: 21 days after Scop treatment, the rats are lightly anesthetized with isoflurane, right heart function is evaluated by echocardiography, and hemodynamic parameters are measured by a right heart catheter.

6. The use of a scullcap-12 in the treatment of pulmonary arterial hypertension according to claim 5, characterized in that, In S13, the rats in the anoxic group and the anoxic+Scop group are subcutaneously injected with SU5416 at a dose of 20mg / kg once a week, and the O2 concentration in the normal-pressure hypoxia incubator is controlled at 10%; the other groups are injected with an equal volume of a DMSO / PEG300 / Tween-80 / normal saline mixture, and the proportion of each component is controlled at 2:8:1:

9.

7. The use of a scullcap-12 in the treatment of pulmonary arterial hypertension according to claim 6, characterized in that, In S2, echocardiography is used to detect heart function, and the specific steps are as follows: S21: after the rats are anesthetized, the rats are fixed on a 37℃ animal operating table in a supine position, and chest hair is removed by depilatory cream; S22: the probe is placed at the root of the parasternal aorta to perform short-axis imaging, and the right ventricular transverse diameter, right atrial transverse diameter, pulmonary artery diameter and aortic diameter are measured; S23: when the apical four-chamber section is used, M-mode ultrasound is used to measure the tricuspid annular plane displacement in the systolic phase; S24: the probe is placed at the root of the parasternal aorta short-axis section at the pulmonary valve to measure the pulmonary valve blood flow spectrum, acceleration time, ejection time and pulmonary valve regurgitation spectrum in the spectrum Doppler mode.

8. The use of a scullcap-12 in the treatment of pulmonary arterial hypertension according to claim 7, characterized in that, In S3, the specific steps are as follows: S31: after the rats are anesthetized, the rats are fixed on a 37℃ animal operating table in a supine position, the right neck is incised, and the right jugular vein is exposed; S32: the jugular vein is cut at a 45° angle with ophthalmic scissors, and the cut is 1 / 3 of the vascular diameter, forming a V-shaped incision of 2-3mm; S33: A polyethylene catheter filled with 0.3% heparin physiological saline was inserted into the right atrium through the port, and the pressure at each site was recorded by an MP150 multi-channel physiological recorder; S34: Keeping the direction unchanged, the catheter continued to advance and rotate counterclockwise so that the head of the catheter hooked the tricuspid valve septum to the left lower, entered the right ventricle, observed the waveform change, and further advanced the catheter to the pulmonary artery to record the pulmonary artery pressure.

9. The use of a scullcap-12 in the treatment of pulmonary arterial hypertension according to claim 8, characterized in that, In S4, the pulmonary arteriole remodeling was observed by HE staining, and the specific steps were as follows: S41: The heart and lung were flushed with physiological saline, the lung tissue was cut, and the blood was removed by fully rinsing with PBS; S42: The lung tissue was soaked in 4% paraformaldehyde and stored at 4°C for 48 hours; S43: Tissue wax block embedding and sectioning were performed for HE staining; S44: The staining condition was observed under an electron microscope and photographed, and 5-8 pulmonary arterioles with a diameter of <100 mm were randomly selected from each slice; S45: The percentage of the pulmonary arteriole media muscle layer to its outer diameter and the percentage of the pulmonary arteriole media muscle layer to the total area of the blood vessel were calculated.

10. The use of a scullcap-12 in the treatment of pulmonary arterial hypertension according to claim 9, characterized in that, In S45, the specific method was to measure the blood vessel outer perimeter and lumen perimeter by Image Pro Plus 6.0 color image analysis system, calculate the blood vessel outer diameter, lumen inner diameter, total blood vessel area and lumen area, and finally calculate the percentage of the pulmonary arteriole media muscle layer to its outer diameter and the percentage of the pulmonary arteriole media muscle layer to the total area of the blood vessel.

Images