The Effects Of Mst1 In Resveratrol Upregulated Autophagy To Postpone Atherosclerosis Development

In recent years,coronary disease has been an increasingly serious global health issue,which produced enormous economic and social burden.In 2012,about 17 million 400 thousand people died of cardiovascular disease,with an estimated 7 million 400 thousand deaths from coronary heart disease,according to WHO.China cardiovascular disease report 2015 shows that cardiovascular disease deaths accounted for the first cause of death of urban and rural residents in the first place,coronary heart disease mortality continues rising.AS is recognized as the pathophysiological basis of cardiovascular and cerebrovascular diseases,for which research of AS is of great significance.Autophagy is a process of self degradation of cell components,which is important for maintaining the homeostasis of cells.A large number of studies have confirmed that autophagy is involved in the pathological process of many diseases,especially in tumor,neurological diseases,metabolic diseases and cardiovascular diseases.Previous studies have shown that autophagy plays an important role in the regulation of the stability of atherosclerosis and atherosclerotic plaque,which provides new opportunities for AS therapy.Resveratrol(RSV)has attracted considerable attention due to its health benefits in cardiovascular disease,cancer,diabetes,and nervous system diseases.Current cell or animal experiments have shown that resveratrol has antioxidant,anti-inflammatory,anti-cancer,cardiovascular protection and protection of the nervous system and other physiological effects.Mst1 is an important component of the Hippo signaling pathway,Hippo signaling pathway is involved in apoptosis and proliferation by regulating the control of organ size and tissue homeostasis.Studies have shown that Mst1 can promote myocardial apoptosis by inhibiting autophagy.In this study,through the establishment of atherosclerosis in mice model to research and discuss the change of autophagy in the process of atherosclerosis,resveratrol intervention to detect its effect on autophagy and atherosclerotic plaque,and detect the change of Mst1 level.At the same time cultured mouse macrophages,establishment of macrophage injury model were observed of resveratrol on macrophage autophagy and apoptosis effect of Mst1 knockdown and overexpression conditions.【Objective】1.To establish AS model and observe the effect of resveratrol on atherosclerotic plaque and the level of autophagy.2.To explore the effect of autophagy in macrophage.3.To investigate the effect of Mst1 on the inhibition of autophagy by inhibiting the development of atherosclerosis.【Methods&Results】1.RSV inhibits atherosclerotic plaque formation in Apo E-/-mice.Apo E-/-mice fed with high fat food to establish atherosclerosis model,Compared with the AS group,serum lipid assay showed that serum triglyceride(TG),total cholesterol(TC),low density lipoprotein cholesterol(LDL-C)of AS+RSV group were significantly decreased(P < 0.05).Compared to the AS group,AS plaque area in the AS+RSV group was statistically significant(P < 0.05),while the AS+RSV+3-MA group decreased the area of autophagy plaque(P < 0.05)Compared with the AS+RSV group.Western blot showed that compared with AS group,the expression of autophagy marker protein LC3 increased and the expression of P62/SQSTM1 decreased in the AS+RSV group,while the expression of Mst1 was down regulated(P < 0.05).Therefore,we suggest that RSV can inhibit the expression of Mst1 by inhibiting the formation of atherosclerotic plaques by regulating autophagy.2.RSV inhibits the formation of macrophage derived foam cells.Cell injury model was established by ox-LDL stimulation of macrophages.Transmission electron microscope showed that,compared with ox-LDL group,the formation of autophagosomes of ox-LDL+RSV group increased(P < 0.05);TUNEL staining results show that,compared with ox-LDL group,ox-LDL+RSV reduced the apoptosis level significantly(P < 0.05).At the same time,the results of Western blot showed that compared with ox-LDL group,the expression level of autophagy marker protein LC3 was increased in ox-LDL+RSV group,the expression of P62/SQSTM1 was decreased significantly,and the expression level of Mst1 protein was decreased overtly(P < 0.05).Therefore,we suggest that resveratrol can increase the level of autophagy,down regulate the level of apoptosis and inhibit the expression of Mst1 in macrophage derived foam cells.3.Resveratrol upregulated autophagy by inhibits Mst1.Mst1 and MOI:100(Ad-sh-Mst1)and Mst1 sh RNA(MOI:100)were used to over express and knock out macrophages Mst1.The results of transmission electron microscope showed that,compared to CON+sh-Lac Z +ox-LDL group,autophagosomes of CON+sh-Mst1 +ox-LDL group was significantly increased(P < 0.05);In addition,CON+sh-Mst1+ox-LDL +RSV group had no significant change of autophagosomes compared with the CON+ sh-Mst1+ox-LDL group.TUNEL staining showed that compared to CON+sh-Lac Z +ox-LDL group,apoptosis level of CON+sh-Mst1 +ox-LDL group decreased significantly(P < 0.05);and compared to the CON+ sh-Mst1+ox-LDL group,CON+sh-Mst1+ox-LDL +RSV group had no significant change on apoptosis level.Western blot results showed that compared to CON+sh-Lac Z +ox-LDL group,CON+sh-Mst1 +ox-LDL group LC3 II expression level increased,P62/SQSTM1 expression decreased obviously(P < 0.05);and compared to the CON+ sh-Mst1+ox-LDL group,CON+sh-Mst1+ox-LDL group,+RSV LC3 II P62/SQSTM1 expression changes were not statistically significant.Therefore,we suggest that: 1.Mst1 inhibits macrophage autophagy.2.RSV inhibits autophagy by inhibiting Mst1.【Conclusion】In this study,we established an AS model with high fat feeding in Apo E-/-mice.Cell injury model was established by ox-LDL stimulation of macrophages.Then,we observed the morphology,blood lipids,protein expressions of AS mice and cell morphology,functions,expressions of macrophages to analyze the effect of RSV on the level of autophagy and the role of Mst1.The results showed that RSV could decrease the level of blood lipid and inhibit the formation of atherosclerotic plaque in AS mice by inhibiting the function of macrophages by Mst1.