| Purpose:To discusse the effect and mechanism of tanshinone ⅡA on hyperlipidemia rats liver lipid deposition at protein level by looking for differentially expressed proteins with i TRAQ technology through hyperlipidemia rat model.Material and method:30 SD rats were randomly divided into the control group(C group),the high-fat diet group(HFD group)and Tanshinone II A treatment group(TAN group)(10 rats per group).The rats in the the HFD group and the TAN group were fed with the high-fat diet.While the C group were fed with the regular balanced diet.At the end of 4 weeks,the rats in TAN group received sodium tanshinone ⅡA sulfonate 10 mg/kg·d by intraperitoneal injection for a further8 weeks.While those in the C group and the HFD group received the same amount of normal saline for the same amount of time.Material was taken after 8 weeks,total cholesterol(TC),total triglyceride(TG),low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C),alanine aminotransferase(ALT)and aspartate aminotransferase(AST)were detected by automatic biochemical analyser.Morphological changes of liver tissue were observed by HE staining.Hepatic lipid deposition were observed by oil red O staining.Liver total protein were extracted and labeled by i TRAQ,then the expressions of differential proteins were detected by mass spectrometric detection and analyzed by software.The expressions of some differential proteins were detected by Western blot.Results:1.The influence of tanshinone ⅡA on the lipid levels and liver function of hyperlipidemia rat.Compared with C group,the levels of TC,TG,LDL-C,ALT and AST levels in HFD group obviously increased(P<0.05,P<0.01),and the level of HDL-C level obviously decreased(P<0.01).Compared with HFD group,the levels of ALT and AST levels decreased(P<0.05,P<0.01)and the level of TC,TG,LDL and HDL have not changed in TAN group.2.The influence of tanshinone ⅡA on the liver pathological morphology and lipid deposition of hyperlipemia rats.HE staining showed : The rats in C group had a normal liver structure,and the liver cells were uniform and morphological structure was clear.The nucleus in C group rats livers was round,clear and middle.HFD group rats liver cell shape was irregular,swelling and had a serious fat vacuoles degeneration.Part of the nucleus boundary was not clear or disappear in HFD group rats liver.The level of cell arrangement was regular,the structure was clear,the degree of edema and cavitation change of cell were reduced,and the nucleus were clear in TAN group rats.Oil red O staining showed: C control group rats liver without lipid of oil red staining.There were lipid drops of Oil red staining significantly increased wich distributed diffuse in HFD group.The lipid drops in the liver cells of TAN group rats were alleviated.3.The influence of tanshinone ⅡA on proteomic expression of hyperlipemia rats liver.Protein quantitative results showed: Compared with C group,there were 485 differential proteins in HFD group,of wich there were 312 proteins up-regulated and 173 proteins down-regulated.Compared with HFD group,there were 252 differential proteins in TAN group,of wich there were 106 proteins up-regulated and 146 proteins down-regulated.The results of intersection analysis of different proteins in three groups showed: There were 61 differential proteins in three group,in witch there were 27 proteins up-regulated in HFD group and down-regulated in TAN group,and 34 proteins down-regulated in HFD group and up-regulated in TAN group.The results of KEGG pathway analysis of different proteins showed: There were 79 pathway were enriched by the proteins expresses up-regulated in HDF group and down-regulated in TAN group,of witch there were 10 path ways p-value<0.05;There were77 pathway were enriched by the proteins expresses down-regulated in HDF group and up-regulated in TAN group,of witch there were 20 path ways p-value<0.05.Among these pathways,there were 5 closed with lipid metabolism: they were arachidonic acid metabolism,fatty acid metabolism,fatty acid degradation,biosynthesis of unsaturate fatty acids and PPAR signaling pathway.4.The influence of tanshinone ⅡA on the expression of differential protein in PPAR signaling pathway of hyperlipemia rats liver.The results of Western Blot showed:Compared with C group,PPAR α、Apoa-I and CYP4A1 proteins expressed down-regulated significantly in HFD group(P<0.01 or P<0.05);Compared with HFD group,PPARα、Apoa-I and CYP4A1 proteins expressed up-regulated significantly in TAN group(P<0.01 or P<0.05).Conclusion:1.Tanshinone ⅡA did not change the serum lipid level of hyperlipidemia rats,but can significantly reduce lipid deposition in hyperlipidemia rats.2.Tanshinone ⅡA can affect the expression of liver proteins in hyperlipidemia rats,and these proteins had closed relationship with fatty acid metabolism,fatty acid degradation,biosynthesis of unsaturate fatty acids,arachidonic acid metabolism and PPAR signaling pathway.3.Tanshinone ⅡA may promote cholesterol transporter and fatty acid beta oxidation and reduce liver lipid deposition by up-regulate the expression of PPARα 、 Apoa-I and CYP4A1. |
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