Effect and its possible mechanism of invigorating spleen and removing phlegm and blood stasis on lipid peroxidation in liver of hyperlipidemia rats
Objective To observe the effect of invigorating spleen and removing phlegm and blood stasis on iron death in liver cells of rats with hyperlipidemia,and to explore its effect and its mechanism on liver lipid peroxidation.Methods Thirty-two SPF SD rats were randomly divided into 4 groups:blank group,model group,Huayu Qutan recipe group and simvastatin group.The rats in the blank group were given normal diet,and the rats in the model group,Huayu Qutan recipe group and simvastatin group were given high fat diet.After 8 weeks,the rats in the Huayu Qutan recipe group and simvastatin group were given intragastric ad-ministration,while the blank group and model group were given the same volume of normal saline.After the last administration,the contents of serum total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDL-C)and high density lipoprotein cholesterol(HDL-C)were detected by automatic biochemical analyzer.The pathological changes of liver were ob-served by hematoxylin-eosin staining;Liver lipid deposition was observed by oil red O staining;the content of iron ion in liver was detected by colorimetry;and the content of ROS and GSH in liver tissue was detected by enzyme-linked immunosorbent as-say(Elisa).The expression of GPx4,SLC7A11,TFR1 mRNA and protein in liver tissue was detected by real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting.Results Compared with the blank group,the contents of serum TC,TG and LDL-C in the model group increased significantly,while the content of HDL-C decreased significantly(P＜0.01).Hepatic steatosis is full of fat vacuoles and hepatocytes become larger.The obvious distribution of red lipid droplets was observed.The con-tents of ROS and Fe2+in liver tissue increased significantly,while the content of GSH decreased significantly(P＜0.01).The ex-pression of TFR1 mRNA and protein in liver tissue increased significantly,while the expression of GPx4 and SLC7A11 mRNA and protein decreased significantly(P＜0.01).Huayu Qutan recipe and simvastatin can significantly reduce the content of TC,TG,LDL-C and increase the content of HDL-C(P＜0.05 or P＜0.01).The degree of steatosis was significantly alleviated and the fatty vacuoles in hepatocytes were significantly reduced.The distribution of lipid droplets decreased significantly.The content of ROS and Fe2+in liver tissue decreased significantly,while the content of GSH increased significantly(P＜0.05 or P＜0.01).The expression of TFR1 mRNA and protein decreased significantly,while the expression of GPx4 and SLC7A11 mRNA and pro-tein increased significantly in liver tissue(P＜0.05 or P＜0.01).Conclusion The mechanism of invigorating spleen and remo-ving phlegm and blood stasis in improving hyperlipidemia rats may be related to the regulation of iron death mediated by SLC7A11/GPx4/ROS signal pathway.
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