Effects Of Yiqi Huoxue Method On Pulmonary Vascular Remodeling In Rats With Hypoxic Pulmonary Hypertension Based On Rho/Rock Pathwa

Purpose:To investigate the effect of invigorating qi and promoting blood circulation on pulmonary vascular remodeling in rats with hypoxic pulmonary hypertension based on Rho/Rock signal pathway.Material and method:1.With 10 rats in each group,fifty male SPF SD rats were divided into blank,model,traditio-nal Chinese medicine,western medicine,and Chinese medicine+Western medicine group s.2.Experiments were conducted out using the drug therapy technique while modeling.With the exception of the blank group,the other four groups of rats were placed in the hypoxic conditions chamber,and the oxygen concentration was managed to keep at（10±1）%.10 hours a day for a total of 28 days.Rats were administered daily 30 min before being placed in the hypoxic chamber.The traditional Chinese medicine group was perfused with 10.61 g·kg^-1·D^-1Yiqi Huoxue Granules;The western medicine group was given 9.45mg·kg^-1·D^-1fasudil hydrochloride injection intraperitoneally;In the group of Chinese medicine+Western medicine,10.61g·kg^-1·D^-1Yiqi Huoxue Granules was given by gavage combined with9.45mg·kg^-1·D^-1fasudilhydrochlorideinjectionintraperitoneally;all three categories were formu-lated into a 2ml drug solution.The blank group and the model group were given the same volume of normal saline by gavage.3.After the experiment,the general conditions of rats were observed.Whether the animal model of hypoxic pulmonary hypertension was successfully established was judged by the mean pulmonary artery pressure（m PAP）,right heart hypertrophy index（RVHI%）and morphological changes of pulmonary arterioles,the percentage of vascular wall thickness in outer diameter（WT%）and the percentage of vascular wall area in total vascular area（WA%）.The level of serum ET-1 was measured by ELISA;The expressions of Rock1 protein,Rho A protein,phosphorylated p-mypt1 and e NOS protein were measured by Western blot;The expressions of Rock1 m RNA and Rho A m RNA were measured by RT-PCR.4.The statistical software SPSS 26.0 was used to analyze the measurement data,and the homogeneity of variance was tested.The one-way ANOVA method was used for one-way ANOVA,and the LSD method was used for pairwise comparison between groups.The measurement data were expressed as mean±standard deviation,and the difference was considered statistically significant with P<0.05.Results:1.The model group’s m PAP,RVHI percent,morphological changes of pulmonary arterioles,a nd indexes of pulmonary arteriole vascular remodeling WT percent and WA percent were sig-nificantly abnormal compared to the blank group（p<0.05or p<0.01）after 28 days ofintermittent hypoxia under normal pressure,indicating that the animal model of HPH was successfully reproduced.2.The general condition of rats in Chinese medicine group,Western medicine group and Chinese medicine+Western medicine group was better.The levels of m PAP,RVHI%,pulmonary artery vascular morphological changes and pulmonary artery vascular remodeling indexes WT%and WA%were significantly improved in the Chinese medicine group,western medicine group and Chinese medicine+Western medicine group compared with the model group（P<0.05 or P<0.01）.There was no statistically significant difference between the Chinese medicine group and the western medicine group（P>0.05）.The efficacy of the Chinese medicine+Western medicine group was better than that of the Chinese and Western medicine groups,and the difference was statistically significant（P<0.05 or P<0.01）.3.Serum ET-1 of rats in each group was detected by ELISA:the level of ET-1 in the model group increased in comparison to the blank group,and the difference was statistically significant（P<0.01）;the level of ET-1 decreased in the other four groups as compared towards the model group,and the difference was statistically significant（P<0.01）;there’s no notable change between the Western medicine group and the Chinese medicine+Western medicine group.4.Western Blot method was used to detect e NOS protein of rats in each group:the expression of e NOS decreased in the rest of the four groups as compared to the blank group,and the thing that is different was statistically meaningful（P<0.01）;because once compared to the model group,the expression of e NOS increased in the Chinese medicine group,the wester n medicine group,and the Chinese medicine+Western medicine group（P<0.01）;Compared with the Chinese medicine group,the expression of e NOS in the Chinese medicine+Western medicine group was increased,and the difference was statistically significant（P<0.01）;there was no significant difference between the expression of e NOS in the Chinese medicine group and the western medicine group（P>0.05）.5.Western blot method was used to detect Rock1,Rho A and p-mypt1 proteins of rats in each group:compared with the blank group,the expressions of Rock1,Rho A and p-mypt1 in the model group increased（P<0.01 or P<0.05）;Compared with the model group,the expressions of Rock1,Rho A and p-mypt1 in traditional Chinese medicine group and Chinese medicine+Western medicine group decreased（P<0.01 or P<0.05）;There was no significant difference in the expression of Rock1,Rho A and p-mypt1 between traditional Chinese medicine group and Western medicine group（P>0.05）.Compared with TCM group,ROCK1 expression in TCM+WESTERN medicine group was decreased（P<0.05）.Compared with the traditional Chinese medicine group and the western medicine group,the expressions of Rho A and P-Myp T1 in the traditional Chinese medicine+Western medicine group were decreased（P<0.01）.6.The Rho A m RNA of rats in each group was detected by RT-PCR:compared with the blank group,Rho A m RNA in model group,traditional Chinese medicine group and Western medicine group increased,and the difference was statistically significant（P<0.01）;Compared with the model group,Rho A m RNA in traditional Chinese medicine group,western medicine group and Chinese medicine+Western medicine group decreased,and the difference was statistically significant（P<0.01）;There was no significant difference in the expression of Rho A m RNA between traditional Chinese medicine group and Western medicine group（P>0.05）;Rho A m RNA in the Chinese medicine+Western medicine group was significantly lower than that in the traditional Chinese medicine group and the western medicine group（P<0.01 or P<0.05）.7.Rock1 m RNA was detected by RT-PCR:compared with the blank group,the Rock1m RNA of the other four groups increased,and the difference was statistically significant（P<0.01 or P<0.05）;Compared with the model group,Rock1 m RNA in traditional Chinese medicine group,western medicine group and Chinese medicine+Western medicine group decreased,and the difference was statistically significant（P<0.01）;There was no significant difference in the expression of Rock1 m RNA between traditional Chinese medicine group and Western medicine group（P>0.05）;Rock1 m RNA in the Chinese medicine+Western medicine group was significantly lower than that in the traditional Chinese medicine group and in the western medicine group(（P<0.01 or P<0.05）.Conclusion:1.Normal pressure intermittent hypoxia can change the behavior of rats,and m PAP,RVHI%,HE staining results and WT%and WA%were significantly changed,indicating that the model of HPH rats was successfully constructed.2.Yiqi Huoxue granules have certain preventive and protective effects on HPH rats,the mechanism may be related to the inhibition of ET-1 expression,the increase of e NOS expression,the protection of vascular endothelial function,thus inhibiting pulmonary vascular remodeling,improving hemodynamic state,and improving right heart function.The regulation of Rho/Rock signaling pathway may inhibit pulmonary vascular remodeling,reduce pulmonary vascular resistance and pulmonary artery pressure,and delay the occurrence and development of disease.3.Chinese medicine and western medicine combined treatment of hypoxic pulmonary hypertension curative effect is better.It reflects the advantages of the combination of Chinese and Western medicine in the treatment of hypoxic pulmonary hypertension,indicating that Chinese medicine also plays a certain role in the prevention and treatment of HPH,providing a new idea and method for the treatment of hypoxic pulmonary hypertension with Chinese medicine.