The Study Of Diosgenin On Monocrotaline Induced Pulmonary Arterial Hypertension In Rats

Objective:The objective of this study was to observe the early application of diosgenin’s effect in PAH rats induced by monocrotaline(monocrotaline,MCT)and to explore its mechanism.Methods: 36 healthy male SD rats,were randomly divided into normal control group(group Control,n=12),MCT model group(group MCT,n=12),pulmonary arterial hypertension diosgenin intervention group(group S,n=12),3 groups rats were feed in the same conditions of feeding standard.Control group: inject the solvent control(ethanol+saline 2:8)in abdominal cavity once,in the 1-28 days after modeling,inject 0.5ml solvent(5%ethanol+distilled water).MCT group: intraperitoneal injection of 1% MCT solution(60mg/kg)once(preparing ethanol and saline according to the volume ratio of 2:8)model,in the 1-28 days after modeling,inject 0.5ml solvent.S group: intraperitoneal injection of 1%MCT solution once(60mg/kg)model,in the 1-28 days after modeling,inject Dio 80mg/kg(dissolved in 0.5ml solvent).According to Sun Bo right heart catheterization method for the determination of mPAP,remove the heart,separate the right ventricular(right ventricular RV)and left ventricular+septum LV+S(left ventricular+septum LV+S),use the paper suck up the moisture,scales to weigh weight,calculate the ratio of RV/LV+S,to determine the degree of right ventricular hypertrophy.HE Chromatids to Observe pulmonary artery structure,measuring the inner,outer diameter,calculating blood vessel wall thickness coefficient D=(vascular outer diameter-lumen inner diameter)* 0.5/outer diameter of vessel,used to evaluate the degree of pulmonary arterioles in membrane thickening.Masson Chromatids to measure proliferation on the pulmonary wall,collagen fiber area(CA),vascular cross-sectional area(CVA)and the ratio of CA/CVA.enzyme-linked immunosorbent,assay,ELISA,detects the level of IL-6 in lung tissue homogenate.Terminal-deoxynucleoitidyl Transferase Mediated Nick End Labeling,TUNEL,detects apoptosis of pulmonary arterial smooth muscle cells,and finally analyzed and compared all data between groups by statistical method.Results: 1.Comparing with Control group,MCT group rat are sign ificantly high(mPAP)、RV/(LV+S)、blood vessel wall thickness coefficient(D)、collagen fiber area(CA)/ vascular cross-sectional area(CVA)(P<0.05),That diosgenin intervenes mPAP、RV/(LV+S)、D、CA/CVA is significantly lower than in MCT group(P < 0.05).2.HE Chromatids Observe pulmonary artery structure in Control Group under light microscope,Pulmonary structure is clear,thin,and consistent,little infiltration of inflammatory substances,MCT pulmonary artery structure is unclear,wall is obvious thickening,the lumen is narrowed,vascular infiltrates a large number of inflammatory cytokines.Comparing with MCT group,the pulmonary artery wall thickness,narrowed lumen,infiltration of inflammatory cytokines significantly lower in S group.3.Masson Chromatids Observe the Control group: Rat pulmonary arteriola has smooth intima,little collagen content,intima,media has a small amount of blue collagen fiber.MCT group: Rat pulmonary arteriola intima disorder,intima,medie has great amount blue collagen fiber,obvious thickening.Comparing with MCT group,Rat pulmonary arteriola intima,media blue collagen fiber is significantly lower in S group.4.ELISA tests and finds that the level of IL-6 in lung tissue homogenate in MCT group is significantly high than in Control group(P< 0.05),in S group was significantly lower than in MCT group(P < 0.05),these results suggest that MCT can induce inflammatory reaction of lung tissue,and Dio significantly inhibited inflammatory reaction induced by the MCT.5.TUNEL method tests and finds that more than staining is positive smooth muscle cells in the Control group,a little positive smooth muscle cells are in the MCT group,more positive smooth muscle cells are in S group.Comparing with Control group,MCT group AI decrease significantly,the difference has statistically significance,Comparing with MCT group,S group’AI significantly reduced,the difference had statistical significance.Conclusion: 1.Dio may prevent rat mPAP、 RV/(LV+S)、 D 、 CA/CVA heightening induced by MCT,suggesting that Dio on MCT-induced rat model of PAH has preventive effect.2.Dio can prevent the inflammatory factors induced by the MCT in lung tissue of rats,in order to prevent their PAH effect mechanism may be related to inhibition of the inflammatory response.3.Dio can promote apoptosis of rat pulmonary arterial smooth muscle cells induced by the MCT,in order to prevent their PAH effect mechanism may be related to promote vascular cells apoptosis.