Study On The Mechanism Of XuefuZhuyu Decoction Regulated Atherosclerotic Vascular Calcification Through Piezo1/NLRP3 Signal Pathway

Objective:Through animal experiments,the therapeutic effect of Xuefu Zhuyu Decoction on atherosclerotic vascular calcification model mice and the expression of Piezo1,NLRP3 and other related genes and proteins were measured to explore the mechanism of Xuefu Zhuyu decoction in reducing vascular inflammation and calcification.This study aimed to provide theoretical support and experimental basis for the treatment of atherosclerotic vascular calcification with Huoxue Huayu prescription.Methods:72 Apo E^-/-mice were randomly divided into control group,model group,atorvastatin group,Xuefu Zhuyu decoction low-dose,medium-dose and high-dose groups.Except for the control group,the other groups were fed high-fat diet for 20 weeks to establish atherosclerotic vascular calcification model.After 20 weeks,two model mice were randomly selected to determine whether the AS model of vascular calcification was successful by alizarin red and silver nitrate staining.After successful modeling,high,medium and low doses of Xuefu Zhuyu decoction and positive control drug atorvastatin were given by intragastric administration for 8 weeks.The general state and body weight of mice were recorded,and the levels of TG,CHOL,HDL-C and LDL-C were detected by automatic biochemical analyzer.HE staining was used to observe the inflammatory infiltration of aorta.Alizarin red and silver nitrate staining were used to detect vascular calcification.The expressions of ALP,BMP4,RUNX2,Piezo1,NLRP3,Casepase1 and IL-1βrelated to aortic calcification were detected by q RT-PCR,immunohistochemistry and western blot.To explore the mechanism of Xuefu Zhuyu Decoction in improving vascular inflammation and calcification in atherosclerotic vascular calcification mice.Results:1.Xuefu Zhuyu Decoction can reduce the weight gain and the increase of serum TG,CHOL and LDL in atherosclerotic vascular calcification mice,and improve the expression level of HDL,with statistically significant differences between groups.2.HE results showed that Xuefu Zhuyu decoction could significantly reduce aortic intimal thickening and inflammatory infiltration in atherosclerotic vascular calcification model mice;The results of alizarin red and silver nitrate showed that Xuefu Zhuyu decoction could effectively reduce the level of aortic calcification in mice with atherosclerotic vascular calcification.3.q RT-PCR results showed that the m RNA expression levels of ALP,BMP4 and RUNX2 were significantly inhibited after administration of Xuefu Zhuyouu Decoction,and the m RNA expression levels of Piezo1 and NLRP3 were significantly decreased in the aorta calcification mice with atherosclerosis.Western blot and immunohistochemical tests showed that Xuefu Zhuyu decoction significantly down-regulated the protein expression of Piezo1 and NLRP3.The results indicated that Xuefu Zhuyu decoction could reduce the inflammation of atherosclerotic vascular calcifications and reverse the development of atherosclerotic vascular calcifications by decreasing the m RNA and protein expression of Piezo1 and NLRP3 in the aorta of atherosclerotic vascular calcifications in mice,and the therapeutic effect was in a dose-dependent manner.Conclusion:Xuefu Zhuyu Decoction can effectively reduce the deposition of calcium salt in aorta,thickening and inflammatory infiltration of vascular intima,and inhibit the development of atherosclerotic vascular calcification in mice with atherosclerotic vascular calcification,which may be related to inhibiting the expression of Piezo1 and NLRP3.