半夏在动脉粥样硬化颈动脉内膜增生中的修复机制研究

Study on Pinellia′s repair mechanism in atherosclerotic carotid intimal hyperplasia

目的研究半夏对动脉粥样硬化大鼠颈动脉内膜增生的修复作用及其潜在机制。方法建立颈动脉损伤内膜增生大鼠模型,随机分为四组,模型组、低剂量半夏组,中剂量半夏组和高剂量半夏组。对各组模型分别使用生理盐水、低[5 g/(m L·kg)]、中[20 g/(m L·kg)]、高[40 g/(m L·kg)]剂量半夏水提液分别进行1、4和8周灌胃干预。通过HE染色观察颈动脉形态,采用Western blotting法检测血管内皮生长因子(VEGF)和内皮型一氧化氮合成酶(e NOS)的表达,采用流式细胞术检测内皮祖细胞(EPCs)的数量变化,采用生化法检测血清中总胆固醇(TCHO)、甘油三酯(TG)和高密度脂蛋白胆固醇(HDL-C)和低密度脂蛋白胆固醇(LDL-C)水平,ELISA法检测血清中白细胞介素6(IL-6)、肿瘤坏死因子-α(TNF-α)、IL-10水平。结果HE染色显示半夏可减轻动脉粥样硬化颈动脉内膜增生。与模型组比较,经半夏干预1周后VEGF、e NOS蛋白表达水平,Dil-ac LDL和FITC-UEA双阳性EPCs细胞比例,TCHO、TG、HDL-C和LDL-C水平,IL-6、TNF-α和IL-10水平比较,差异均无统计学意义(P>0.05);经半夏干预4周和8周后,VEGF和e NOS蛋白表达水平,Dil-ac LDL和FITC-UEA双阳性EPCs细胞比例,HDL-C、IL-10水平显著上调;TCHO、TG、LDL-C、IL-6、TNF-α水平显著下调,差异有统计学意义(P<0.05);均呈时间和剂量效应。结论半夏可通过增加EPCs数量来抑制动脉粥样硬化大鼠颈动脉的内膜增生,同时降低动脉粥样硬化大鼠的血脂和炎症水平。

Objective To study the repairing effects of Pinellia on carotid intimal hyperplasia in atherosclerotic rats and its potential mechanism.Methods The carotid artery intimal hyperplasia rats model was established and randomly divided into 4 groups:model group,low-dose Pinellia group,medium-dose Pinellia group and high-dose Pinellia group.Each group was administered with saline(model group),low-dose[5 g/(mL·kg)],medium-dose[20 g/(mL·kg)]),high-dose[40 g/(mL·kg)]Pinellia water extract for 1 week,4 weeks and 8 weeks.The carotid artery morphology was observed by HE staining;the expressions of vascular endothelial growth factor(VEGF)and endothelial nitric oxide synthase(eNOS)were detected by Western blotting;the number of endothelial progenitor cells(EPCs)was detected by flow cytometry.The serum levels of total cholesterol(TCHO),triglycerides(TG),high density lipoprotein cholesterol(HDL-C)and low density lipoprotein cholesterol(LDL-C)were determined by biochemical methods.The serum interleukins 6(IL-6),tumor necrosis factor-α(TNF-α)and IL-10 levels were determined by ELISA.Results HE staining showed that Pinellia could reduce atherosclerotic carotid artery intimal hyperplasia.Compared with the model group,the expression levels of VEGF and eNOS protein,the proportion of Dil-acLDL and FITC-UEA double-positive EPCs,the levels of TCHO,TG,HDL-C and LDL-c,IL-6,TNF after 1 week of intervention with Pinellia in the treatment groups showed no significant differences(P>0.05).After 4 and 8 weeks of Pinellia intervention,the expression levels of VEGF and eNOS protein,the proportion of Dil-acLDL and FITC-UEA double-positive EPCs,levels of HDL-C,IL-10,TCHO,TG,LDL-C,IL-6 and TNF-αin the treatment groups were significantly different from those in the model group(P<0.05),while HDL-C,IL-10 levels were significantly up-regulated and TCHO,TG,LDL-C,IL-6 and TNF-αlevels were significantly down-regulated;all showed time and dose effects.Conclusion Pinellia could inhibit the intimal hyperplasia of carotid arteries in atherosclerotic rats through increasing the number of EPCs,and reduce the blood lipid and inflammation levels of atherosclerotic rats.