Based On The MTOR Signal Pathway, To Explore The Effect Mechanism Of Bushen Jiangzhi Prescription Regulating Autophagy And Interfering With Atherosclerosis In ApoE^-/- Mice

OBJECTIVE:To explore the relevant mechanism of Bushen Jiangzhi Recipe in the treatment of AS by regulating the expression of key factors in mTOR signaling pathway and promoting autophagy.METHODS:In this study,96 male apoE^-/-mice of 12 weeks old fed with high-fat diet were randomly divided into 4 groups by random number table:apoE^-/-mice+high-fat diet control group?Model group,n=24?,apoE^-/-mice+high-fat diet+Bushen Jiangzhi Recipe group?Bushen group,n=24?,apoE^-/-mice+high-fat diet+atorvastatin group?Statin group,n=24?,apoE^-/-mice+high-fat diet+3-methyladenine group?3-MA group,n=24?,same age,same strain male C57BL/6 mice as normal control group?Normal group,n=24?.The pathological morphology,lipid accumulation,collagen fibers and elastic fibers of mouse aortic sinus were observed by HE,Oil Red O,Masson and EVG staining.The aortic aging phenotype of mice was detected by?-GAL staining.The autophagosomes of mouse aorta were observed by transmission electron microscope.Serum lipids?TC,TG,HDL-C,LDL-C?and inflammatory factors?TNF-?,IFN-?,IL-10?were detected by ELISA.The expression of mTOR,Akt,s6k1,LC3,Beclin 1,P62,P21 and P16 protein in mouse aorta was detected by Western Blot.The expression of mTOR and P62 protein in mouse aortic sinus was detected by immunohistochemistry.RESULTS:High-fat diet induced apoE^-/-mouse aortic sinus can form typical AS plaque,lipid accumulation,collagen fiber and elastic fiber content decreased;?-GAL staining showed that the aorta positive staining area of the model group increased.Transmission electron microscopy showed that aortic autophagosomes decreased;aortic mTOR,Akt,s6k1,P62,P21,P16 protein expression and serum TNF-?,IFN-?,TC,LDL-C levels were significantly increased,LC3,Beclin1 protein expression and serum IL-10 The HDL-C content is significantly reduced.3-MA could further aggravate the atherosclerotic lesion of aortic sinus in apoE^-/-mice,increase the area of atherosclerotic plaque,lipid accumulation and beta-GAL positive staining of aortic sinus,reduce the content of collagen fibers and autophagic bodies in aortic sinus,and reduce the elastic fibers of aortic sinus vessels.It could significantly reduce the content of LC3,Beclin1 protein in aorta of apoE^-/-mice fed with high-fat diet and serum IL-10 in peripheral blood.The expression of P62,P21 and P16 protein in aorta and serum levels of TC,LDL-C,TNF-?and IFN-?in mice were elevated.Bushen Jiangzhi Recipe can significantly improve aortic sinus AS lesions induced by high-fat diet in apoE^-/-mice,reduce AS plaque area and lipid accumulation,increase collagen fiber and elastic fiber content;?-GAL staining showed a reduction in aortic positive staining area;Increased aortic autophagy bodies;Bushen Jiangzhi Recipe can also decrease the expression of mTOR,Akt,s6k1,P62,P21,P16 protein and serum TNF-?,IFN?,TC and LDL-C in mouse aorta,increase LC3,Beclin1 protein and serum IL-10,HDL-C content.CONCLUSION:Bushen Jiangzhi Recipe may effectively treat AS by regulating the expression of key factors in mTOR signaling pathway and promoting autophagy.