| Purpose: By observing the effect of Xiangsha Liujunzi Tang on the expression of related genes and proteins in the dysfunctional high density lipoprotein(dy HDL)and reverse cholesterol transport(RCT)pathway in rats with spleen deficiency hyperlipidemia,taking dy HDL as an entry point,we explore the mechanism of improving blood lipid disorder and liver lipid deposition in rats with spleen deficiency hyperlipidemia.Material and method: Seventy-five SPF SD rats were randomly divided into the normal control group,the high fat group,spleen deficiency and high fat group,the Xiangsha Liujunzi low dose group(5.67 g·kg-1·d-1)and the Xiangsha Liujunzi high dose group(11.34g·kg-1·d-1).In the spleen deficiency and high fat group,the Xiangsha Liujunzi low dose group and the Xiangsha Liujunzi high dose group,the composite method of diet and exhaustively swimming was used for 15 days,and normal group was fed with basic diet,the high-fat group,spleen-deficiency and high-fat group,the Xiangsha Liujunzi low dose group and the Xiangsha Liujunzi high dose group were fed with high-fat diet.Free food intake,unlimited water.Animal samples were collected after 14 weeks of feeding with high-fat diet,4 weeks of drug intervention before animal samples were collected,,the Xiangsha Liujunzi low dose group and the Xiangsha Liujunzi high dose group were given the corresponding doses,the normal group,the high fat group and spleen deficiency high fat group were given the corresponding volume of normal saline.The content of total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol cholesterol(LDL-C),high density lipoprotein cholesterol(HDL-C)were detected by automatic biochemistry analyzer;The D-xylose excretion rate was measured by phloroglucinol method;The morphological changes of liver cells were observed by HE staining;The level of PON1?apo A1?SAA of blood plasma and PLTP?LCAT?CETP of blood serum were detected by ELISA;The expression of gene m RNA of PON1?apo A1?SAA?PLTP?LCAT and CETP in rat liver was detected by Realtime Q-PCR;The expression of protein of PON1?apo A1?SAA?PLTP?LCAT?CETP and CYP7A1 in rat liver was detected by Western blotting.Results:1.Effect of Xiangsha Liujunzi Tang on blood lipid level in rats with spleen-deficiency hyperlipidemiaCompared with the normal control group,the serum TC,TG and LDL-C in the high-fat group and the spleen deficiency and high fat group were significantly increased(P<0.05 or P<0.01),and the HDL-C was significantly reduced(P<0.01).The changes in blood lipids in rats with spleen-deficiency hyperlipidemia were significantly larger than those in the high-fat group,but there was no statistical significance.Compared with the spleen deficiency and high-fat group,the serum TC,TG and LDL-C in the low-dose and high-dose groups of Xiangsha Liujunzi Tang were significantly reduced(P<0.01),and HDL-C increased significantly(P<0.01),There was no obvious dose-effect relationship between low dose group and the high dose group of Xiangsha Liujunzi.2.Changes in urine D-xylose excretion rate of rats in various groups and the intervention effect of Xiangsha Liujunzi TangCompared with the normal control group,the urine D-xylose excretion rate of rats in the spleen deficiency and high-fat group was significantly reduced(P<0.01).After the intervention of Xiangsha Liujunzi Tang,the urine D-xylose excretion rate in rats was significantly increased(P<0.05),Among them,the urine D-xylose excretion rate of rats in the high dose group of Xiangsha Liujunzi was significantly increased.3.Effect of Xiangsha Liujunzi Tang on pathological morphology of liver tissue in rats with spleen deficiency and hyperlipidemiaThe hepatocytes in the normal control group were round,large and centered,with clear cell lines and normal hepatic sinus.Compared with the normal control group,hepatocytes in the high-fat group and spleen deficiency and high-fat group became enlarged and rounded,with a large number of fat vacuoles and abnormal liver cell morphology,the cytoplasm was loose,part of the nucleus squeezed into the cell membrane;and compared with the high-fat group,fat vacuoles in liver cells of the spleen deficiency and high-fat group were significantly increased and liver cells were abnormal;After the intervention of Xiangsha Liujunzi Tang,Rat liver cell fat The vacuoles were significantly reduced,the hepatocytes were less swollen,the cells arranged neatly and clearly,and the nuclei were normal,Compared with the low dosegroup of Xiangsha Liujunzi,the liver cell status of rats in the high dose group of Xiangsha Liujunzi improved more obviously.4.Effect of Xiangsha Liujunzi Tang on Content of PON1?apo A1?SAA of blood plasma and PLTP?LCAT?CETP of blood serumCompared with the normal control group,the contents of SAA of blood plasma,the contents of PLTP and CETP of blood serum in rats with high fat group and spleen deficiency and high fat group were significantly increased(P<0.01),and the contents of PON1 and apo A1 of blood plasma were significantly reduced(P<0.01),changes of content of LCAT of blood serum showed a downward trend,LCAT of blood serum and the PON1 and SAA contents of blood plasma in the blood of rats with spleen deficiency and high fat group were significantly lower than those in high fat group(P<0.05 or P<0.01),and the contents of PLTP and CETP of blood serum were significantly higher than those in high fat group(P<0.05);After the intervention of Xiangsha Liujunzi Tang in the hypertensive rats,the contents of SAA of blood plasma,the contents of PLTP and CETP of blood serum in rats were significantly reduced(P<0.01),and the contents of PON1 and apo A1 of blood plasma,LCAT of blood serum were significantly increased(P<0.01),Among them,the content of plasma dy HDL and serum RCT pathway related proteins in the high dose group of Xiangsha Liujunzi was more obvious.5.Effect of Xiangsha Liujunzi Tang on expression of gene m RNA of PON1?apo A1?SAA?PLTP?LCAT and CETP in liverCompared with the normal control group,the levels of SAA,PLTP and CETP m RNAs in the liver of rats with high fat group and spleen deficiency and high fat group were significantly increased(P<0.01),and the levels of PON1,apo A1 and LCAT genes were significantly reduced(P<0.05 or P<0.01).And the level of PLTP gene m RNA in liver of rats with spleen deficiency and high fat group was significantly lower than that in high fat group(P<0.05).After the intervention of Xiangsha Liujunzi Tang,the levels of SAA,PLTP and CETP gene m RNA in liver of rats were significantly reduced(P<0.05 or P<0.01),m RNA levels of apo A1 and LCAT genes were significantly increased(P<0.05 or P<0.01),m RNA levels of PON1 gene showed an upward trend,but no statistical significance.Among them,the expression changes of dy HDL and RCT-related gene m RNA in the liver of XiangshaLiujunzi high dose group was more obvious.6.Effect of Xiangsha Liujunzi Tang on expression of protein expression of PON1?apo A1?SAA?PLTP?LCAT?CETP and CYP7A1 in liverCompared with the normal control group,the relative expression levels of SAA and PLTP protein in liver of rats with high fat group and spleen deficiency and high fat group were significantly increased(P<0.05),and the relative expression levels of apo A1,LCAT and CYP7A1 proteins Significantly decreased(P<0.05 or P<0.01),the relative expression level of CETP protein in liver of rats with spleen deficiency and high fat group was significantly increased(P<0.05);After the Xiangsha Liujunzi Tang intervention,the relative expression levels of PON1 and CYP7A1 proteins in the liver of the low dose group of Xiangsha Liujunzi Tang significantly increased(P<0.05),while the relative expression levels of PLTP protein significantly decreased(P<0.05).The relative expression levels of PON1,apo A1,LCAT,and CYP7A1 proteins in the liver of rats in Xiangsha Liujunzi high dose group were significantly increased(P<0.05),and the relative expression levels of SAA proteins were significantly reduced(P<0.05).Conclusion:1.Xiangsha Liujunzi Tang can effectively regulate dyslipidemia in rats with spleen-deficiency hyperlipidemia and liver foaming state2.Xiangsha Liujunzi Tang may play a role in correcting lipid disorders by affecting the expression of relevant genes and proteins in the dy HDL and pathways of RCT. |
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