The Effect Of Gualou Xiebai Banxia Decoction On Lipid Metabolism And Expression Of Lox-1 In ApoE^-/- Mice With Atherosclerosis

Objective:To observe the effects of Gualou Xiebai Banxia Decoction on lipid metabolism?TC,TG,LDL-C,HDL-C?,oxidative stress?SOD,MDA,GSH-px?,ox-LDL and Lox-1 in ApoE^-/-mice with atherosclerosis,and possible mechanism was explored.Method:1.Establishment of ApoE^-/-mice model with atherosclerosis?AS?42 ApoE^-/-and 12 C57BL/6J male mouse were fed with normal diet in laboratory.One week later,ApoE^-/-male mouse were fed with high fat diet and C57BL/6J male mouse were fed with normal diet.Then the two were selected from Apo-E^-/-mouse and C57BL/6J mouse randomly after 8 weeks.Blood samples were drawn from their orbital vein to test blood lipid levels,and observed the pathological changes by HE staining,that determined the establishment of AS model.2.Group and administrationAS model mouse were randomly divided into GXBD high dose group(GXBD-H,20 g·kg^-1)and GXBD low dose group(GXBD-L,10 g·kg^-1),simvastatin group(XFTT,5 mg·kg^-1)and the model group?Model?.C57BL/6J mice was used as the normal control group?Normal?.Each group of mouse was orally administered according to the corresponding dose,once a day for 8weeks.The Model group and the Normal group were oral administration with an equal volume of saline.The mouse were weighed and anesthetized at 24 h after the last administration.Blood samples were drawn from mice orbital vein,and took supernatant after centrifugation with speed of 3000 r·min^-1,stored at 20?.3.Detection indicator3.1 The serum levels of TC,TG,LDL-C,and HDL-C in ApoE^-/-mice were measured by an automatic biochemical analyzer.3.2 The pathological changes of aorta in ApoE^-/-mice were observed by HE staining.3.3 The content of ox-LDL in serum of ApoE^-/-mice was detected by ELISA.3.4 The activity of SOD in serum of ApoE^-/-mice was detected by the hydroxylamine method.The content of MDA was detected by TBA method.The content of GSH-px was detected by colorimetry.3.5 The expression of Lox-1 mRNA in aortic tissue of ApoE^-/-mice was detected by RT-qPCR.3.6 The expression of Lox-1 protein in aortic tissue of ApoE^-/-mice was detected by Western Blot.Result:1.Effect of GXBD on blood lipid levels in ApoE^-/-miceCompared with the Normal group,the levels of TC,TG,and LDL-C in the model group increased,and the HDL-C level decreased?P<0.05?,which was demonstrated the mice model was successfully established.Compared with the Model group,levels of TC,TG,and LDL-C in the GXBD-L,GXBD-H,and XFTT group decreased?P<0.05?,and HDL-C levels increased?P<0.05?.2.The effect of GXBD on the pathological changes of aorta in ApoE^-/-miceNormal group:The aortic intima was intact and smooth,and the endothelial cells were fold in fold.Model group:The blood vessel wall was partially damaged,atherosclerotic plaques formed in the blood vessels,there were abundant lipids in the plaques,some inflammatory cells infiltrated around the plaques,and there are a lot of foam cells under the fiber caps.Compared with Model group,GXBD-L group less damaged the inner wall of aorta,amounts of foam cells,plaque area and lipid contents decreased,and there was a small amount of foam cells and inflammatory cells in plaques.In the GXBD-H group and the XFTT group,the inner wall of the blood vessel was relatively smooth and the inner membrane was damaged occasionally.There was less foam cell aggregation and inflammatory cell infiltration under the microscope.3.The effect of GXBD on the content of ox-LDL in serum of ApoE^-/-miceCompared with the Normal group,the level of ox-LDL in the plasma of Model mice increased?P<0.05?.Compared with the Model group,the levels of ox-LDL in the plasma of GXBD-H,GXBD-L and XFTT mice were reduced in varying degrees?P<0.05?.Compared with the GXBD-L group,the serum levels of ox-LDL in XFFT and GXBD-H group were decreased?P<0.05?.Compared with the high dose group of GXBD,the level of ox-LDL in the serum of XFTT group was decreased?P<0.05?.4.The effect of GXBD on the content of SOD,GSH-px and MDA in serum of miceCompared with the Normal group,the level of MDA in the plasma of Model mice increased and the level of SOD and GSH-px decreased?P<0.05?.Compared with the Model group,the level of MDA in the plasma of GXBD-H,GXBD-L and XFTT mice decreased in varying degrees,and the level of SOD and GSH-px increased?P<0.05?.Compared with GXBD-L,MDA level in group GXBD-H and XFTT decreased?P<0.05?,GSH-px level increased?P<0.05?,SOD level increased?P>0.05?,and compared with GXBD-L group,levels of MDA,SOD and GSH-px in XFTT group increased?P>0.05?.5.The effect of GXBD on the expression of Lox-1 mRNA in the aorta of ApoE^-/-miceCompared with the Normal group,the expression of Lox-1 mRNA in the aorta tissue of the Model group increased?P<0.05?.Compared with the Model group,the expression level of Lox-1 mRNA in the aorta of GXBD-H,GXBD-L and XFTT mice decreased in varying degrees?P<0.05?.Compared with GXBD-L group,the expression level of Lox-1 mRNA in aortic tissue of GXBD-H and XFTT group decreased?P<0.05?,compared with GXBD-H group,the expression level of Lox1 mRNA in XFTT group decreased?P<0.05?.6.The effect of GXBD on the expression of Lox-1 protein in the aorta of ApoE^-/-miceCompared with the Normal group,the expression of Lox-1 protein in the aorta tissue of the Model group increased?P<0.05?.Compared with the Model group,the level of Lox-1 protein in the active tissues of GXBD-H,GXBD-L and XFTT mice decreased in varying degrees?P<0.05?.Compared with GXBD-L group,the expression level of Lox-1 protein in aortic tissue of GXBD-H and XFTT group decreased?P<0.05?.Compared with GXBD-H group,Lox-1 mRNA expression level in aorta tissue of XFTT group decreased?P<0.05?.Conclusion:In this study,the AS model of ApoE^-/-mice was established,and GXBD and simvastatin were oral administration to compare results,which showed that GXBD can significantly improve lipid metabolism and oxidative stress on AS mice.It played the role of prevention and treatment of AS in many ways.The possible mechanism of action is as follows:1.GXBD can inhibit the development of atherosclerosis by regulating blood lipid metabolism,increasing HDL levels and decreasing LDL levels.2.GXBD can inhibit lipid peroxidation by strongthening the antioxidant capacity of AS mice.3.GXBD can reduce the levels of ox-LDL in serum on AS mice and delay the progression of atherosclerosis.4.GXBD can reduce the levels of Lox-1 in aortic tissue,inhibit oxidative stress injury mediated by Lox-1 and induced by ox-LDL,which is a possible mechanism for delaying the development of atherosclerosis.