| Objective:By observing the related mechanism of the different combinations of Astragalus membranaceus in Buyang Huanwu Decoction regulating the molecular and protein level of Rho kinase signaling pathway,to explore the effect of specific Chinese medicine dosage differences on atherosclerosis in the Rho kinase signaling pathway,so as to clarify the target and pathway of Chinese medicine compound anti-AS and provide relevant evidence for the further application of traditional Chinese medicine prescriptions in the prevention and treatment of AS.Methods:1.The AS rat model was replicated with high-fat diet combined with vitamin D3.After the model was successfully replicated,it was treated with Buyang Huanwu Decoction of different ratio of Astragalus.2.After the intervention of Buyang Huanwu Decoction,the whole aorta was taken,and the pathological changes of the aortic tissue of the rats in each group were detected.The histomorphological changes of the aorta in different treatment groups were observed.3.Blood was taken from the abdominal aorta,and the levels of serum total cholesterol(TC),triglycerides(TG),high-density lipoprotein cholesterol(HDL-C)and low-density lipoprotein cholesterol(LDL-C)were determined by enzymatic method.The changes of biochemical indexes were analyzed and the intervention effect of Buyang Huanwu decoction was observed.4.Western Blot method was used to determine the expression of ROCK1,ROCK2,p-MYPT-1,p-MLCP and p-MLC in the aorta,to evaluate the changes in Rho kinase activity and the effect of Rho kinase on p-MYPT-1,p-MLCP and p-MLC.The intervention effect of Buyang Huanwu decoction was observed.Results:1.Compared with the blank control group,the levels of TC,TG,and LDL-C in the model control group were significantly increased,and the levels of HDL-C were significantly reduced,and the differences were statistically significant(P<0.05).Compared with the model control group,the levels of TC,TG,and LDL-C in all treatment groups were significantly reduced,and the level of HDL-C was significantly increased.Among them,the Astragalus 120 g group reduced TC,TG,LDL-C and increased HDL-C levels better than other Astragalus different ratio of Buyang Huanwu Decoction treatment group,the difference was statistically significant(P<0.05).There was no significant difference from the simvastatin treatment group(P>0.05).2.Under the light microscope,the model control group can observe the media atrophy of the abdominal aorta,disordered smooth muscle arrangement,a large amount of calcium deposits under the obvious fibrous cap,and a large area of inflammatory cell infiltration and partly hyperplastic connective tissue can be seen.Compared with the model control group,the pathological changes of AS in all treatment groups were improved to varying degrees.Among them,the abdominal aorta of rats in the Astragalus 30 g group showed damage to the vascular endothelial structure,and some areas had atherosclerotic plaque formation and inflammatory cell infiltration;the abdominal aorta of rats in the Astragalus 60 g group had a relatively unclear vascular structure.The smooth muscle cells of the media are arranged disorderly,and some atheromatous plaques can be seen;In the astragalus 90 g group,the smooth muscle cells of the intima of the abdominal aorta of rats proliferated,and a lipid nucleus was formed between the smooth basal layer of the intima and media;the vascular structure of the abdominal aorta of rats in the Astragalus 120 g group was normal and clear,and there was a tendency for atherosclerosis in some areas;the vascular wall structure of the abdominal aorta of rats in the simvastatin group was relatively clear and normal,with a small amount of foam cells visible,and atherosclerotic plaques formed in some areas or had formed atherosclerotic plaques.3.Compared with the blank control group,the expression of ROCK1 protein in the model control group,Astragalus 30 g group,Astragalus 60 g group,and Astragalus 90 g group increased,and the difference was statistically significant(P<0.05).Compared with the model control group,the expression of ROCK1 protein in the Astragalus 60 g group,Astragalus 90 g group,Astragalus 120 g group and Simvastatin group decreased,and the difference was statistically significant(P<0.05).Astragalus 30 g group protein expression decreased,but the difference was not statistically significant(P>0.05).Compared with the other Buyang Huanwu Decoction treatment groups,the Astragalus 120 g group showed a decrease in ROCK1 protein expression,and the difference was statistically significant(P<0.05).Compared with the Simvastatin treatment group,the 120 g group protein expression was reduced,but the difference was not statistically significant(P>0.05).Compared with the astragalus 30 g group,the astragalus 90 g group and simvastatin group protein expression were significantly reduced,the difference was statistically significant(P<0.05);the other differences were not statistically significant.4.Compared with the blank control group,the expression of ROCK2 protein in the model control group,Astragalus 30 g group and Astragalus 60 g group increased,and the difference was statistically significant(P<0.05).There was no significant change in the ROCK2 protein expression of 90 g astragalus group,120 g astragalus group and simvastatin group,and the difference was not statistically significant(P>0.05).Compared with the model control group,ROCK2 protein expression in the Astragalus 30 g group,Astragalus60 g group,Astragalus 90 g group,Astragalus 120 g group and Simvastatin group was reduced,and the difference was statistically significant(P<0.05).Compared with other groups treated with Buyang Huanwu Decoction,the Astragalus 120 g group showed a decrease in ROCK2 protein expression,and the difference was statistically significant(P<0.05).Compared with the Astragalus 120 g group and the Simvastatin treatment group,although the ROCK2 protein expression was reduced,the difference was not statistically significant(P>0.05).There was no significant difference in ROCK2 protein expression in the Astragalus 30 g group,Astragalus 60 g group,Astragalus 90 g group and Simvastatin group,and the difference was not statistically significant(P>0.05).5.Compared with the blank control group,the expression of p-MYPT-1 protein in the model control group,Astragalus 30 g group,Astragalus 60 g group,and Astragalus 90 g group increased,and the expression of Astragalus 120 g protein decreased,and the difference was statistically significant(P<0.05),Simvastatin protein expression did not change significantly,and the difference was not statistically significant(P>0.05).Compared with the model control group,the expression of p-MYPT-1 protein in the Astragalus 30 g group,Astragalus 60 g group,Astragalus 90 g group,Astragalus 120 g group and Simvastatin group decreased,and the difference was statistically significant(P<0.05).Compared with other treatment groups,the expression of p-MYPT-1 protein in the astragalus 120 g group was reduced,and the difference was statistically significant(P<0.05);compared with the astragalus 30 g group,the astragalus 60 g group,the astragalus 90 g group,the astragalus 120 g group and Simvastatin group protein expression was significantly reduced,the difference was statistically significant(P<0.05);compared with the Astragalus 60 g group,the simvastatin group expression of p-MYPT-1protein was reduced,the difference was statistically significant(P<0.05);the other differences were not statistically significant.6.Compared with the blank control group,the expression of p-MLCP protein increased in the model control group,Astragalus 30 g group,Astragalus 60 g group,Astragalus 90 g group and Simvastatin group,and the difference was statistically significant(P<0.05),the expression of Astragalus 120 g protein did not change significantly,the difference was not statistically significant(P>0.05).Compared with the model control group,the expression of p-MLCP protein in the Astragalus 30 g group,Astragalus 60 g group,Astragalus 90 g group,Astragalus 120 g group and Simvastatin group was decreased,and the difference was statistically significant(P<0.05).Compared with other treatment groups,the expression of p-MLCP protein in the Astragalus 120 g group was reduced,and the difference was statistically significant(P<0.05);Compared with the Astragalus 30 g group,simvastatin protein expression was significantly reduced,and the difference was statistically significant(P<0.05);the other differences were not statistically significant.7.Compared with the blank control group,the expression of p-MLC protein in the model control group,Astragalus 30 g group,Astragalus 60 g group,and Astragalus 90 g group increased,and the difference was statistically significant(P<0.05).The astragalus120 g group and simvastatin protein expression did not change significantly,and the difference was not statistically significant(P>0.05).Compared with the model control group,the expression of p-MLC protein in the Astragalus 30 g group,Astragalus 60 g group,Astragalus 90 g group,Astragalus 120 g group and Simvastatin group decreased,and the difference was statistically significant(P<0.05).Compared with the other treatment groups,the Astragalus 120 g group had reduced p-MLC protein expression,and the difference was statistically significant(P<0.05);compared with the Astragalus 60 g group,simvastatin protein expression was significantly reduced,and the difference was statistically significant(P<0.05);the other differences were not statistically significant.Conclusion:1.Buyang Huanwu Decoction can significantly reduce the levels of TC,TG,LDL-C and increase the level of HDL-C in AS model rats,indicating that AS can be treated by regulating blood lipid levels.2.Buyang Huanwu Decoction can improve the damage of the aortic vascular endothelial cells of AS model rats,reduce the formation of foam cells,and reduce the inflammation of the intima and the degree of plaque sclerosis.3.Buyang Huanwu Decoction can regulate the Rho kinase signaling pathway by inhibiting the expression of ROCK1 and ROCK2 and phosphorylation of MLC,so as to achieve the effect of treating AS.4.During the treatment of AS model rats with Buyang Huanwu Decoction with different ratio of Astragalus,Buyang Huanwu Decoction with Astragalus 120 g content played the most significant role in all aspects.It can provide a reference for the prescription to choose the best combination. |
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