黄芪熊竹素通过NOX4/ROS/NF-κB通路抑制血管紧张素Ⅱ诱导的血管平滑肌细胞增殖  

Jaranol Attenuating AngⅡ-induced HA-SMCs Proliferation by Inhibiting NOX4/ROS/NF-κB Pathway

作　　者：张铭杰 刘晶[1] 聂慧娟[2] 刘天龙[1] 肖志彬 Zhang Mingjie;Liu Jing;Nie Huijuan;Liu Tianlong;Xiao Zhibin(Department of Pharmacy,Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010030,China;Department of Cardiology,Affiliated Hospital of Inner Mongolia Medical University;Department of Clinical Pharmacy,College of Pharmacy,Inner Mongolia Medical University)

机构地区：[1]内蒙古医科大学附属医院药剂部,呼和浩特010030 [2]内蒙古医科大学附属医院心内科 [3]内蒙古医科大学药学院临床药学教研室

出　　处：《中国药师》2022年第10期1697-1702,共6页China Pharmacist

基　　金：国家自然科学基金项目(编号:81960048、82160058);内蒙古自治区自然科学基金项目(编号:2019BS08003、2021LHMS08043);内蒙古医科大学百万工程项目[编号:YKD2018KJBW(LH)0026];内蒙古医科大学附属医院博士启动金项目(编号:NYFY BS 202114)。

摘　　要：目的:本研究旨在评价熊竹素(JAR)对血管紧张素Ⅱ(AngⅡ)诱导的血管平滑肌细胞增殖的抑制作用及机制。方法:体外培养人主动脉平滑肌细胞株(HA-SMCs),10^(-6) mol·L^(-1)的AngⅡ诱导HA-SMCs增殖并使用梯度稀释的JAR进行干预,检测HA-SMCs活性、培养基中过氧化氢酶(CAT)、超氧化物歧化酶(SOD)及谷胱甘肽过氧化物酶(GSH-Px)活力。使用2′,7′-二氯荧光素二乙酸酯(DCFH-DA)探针和流式细胞仪检测HA-SMCs中ROS水平。利用实时荧光定量聚合酶链式反应(RT-PCR)和蛋白质印迹法(Western blot)检测HA-SMCs中尼克酰胺腺嘌呤二核苷酸磷酸氧化酶(NOX)4、NOX2及核因子κB(NF-κB)的表达水平。结果:JAR对AngⅡ诱导的HA-SMCs活性增加具有显著抑制作用(P<0.01)。AngⅡ诱导组HA-SMCs中荧光强度较对照组细胞显著增加(P<0.05或P<0.01),说明细胞中ROS水平较对照组显著增加,而JAR干预能够显著地降低AngⅡ诱导的ROS生成(P<0.05)。AngⅡ诱导组HA-SMCs培养基中CAT、SOD及GSH-Px活力较对照组显著降低(P<0.01),而JAR干预组上述酶活力显著增加(P<0.05或P<0.01)。RT-PCR检测结果显示,AngⅡ组NOX4和NOX2在HA-SMCs中的表达较对照组显著升高(P<0.05或P<0.01),而JAR干预能够显著抑制AngⅡ诱导的NOX4和NOX2在HA-SMCs中的表达(P<0.05)。Western blot检测结果显示,AngⅡ诱导HA-SMCs 48 h后,NOX4、NOX2及p-NF-κB在HA-SMCs中的表达水平显著增加(P<0.01),而JAR干预能够显著抑制上述蛋白的表达(P<0.05)。结论:黄芪熊竹素通过NOX4/ROS/NF-κB通路抑制AngⅡ诱导的血管平滑肌细胞增殖。Objective:To evaluate the inhibitory effect of ianarol(JAR)on angiotensinⅡ(AngⅡ)-induced HA-SMCs proliferation.Methods:10^(-6) mol·L^(-1) AngⅡwas used to induce HA-SMCs proliferation in vitro,which was treated with jaranol(JAR).HA-SMCs viability,and enzyme activities of CAT,SOD and GSH-Px in medium were detected by kits.DCFH-DA probe and flow cytometer were used to detect ROS level in HA-SMCs.Expression levels of NOX4,NOX2 and NF-κB in HA-SMCs were detected by real time-PCR(RT-PCR)and Western blot.Results:The results showed that JAR had a significantly inhibitory effect on AngⅡ-induced HA-SMCs proliferation(P<0.01).The level of ROS in cells was significantly higher than that in the control group(P<0.05 or P<0.01),and JAR intervention could significantly reduce the production of ROS induced by AngⅡ(P<0.05).Significant increase in enzyme activities of CAT,SOD and GSH-Px was found in AngⅡ-induced HA-SMCs(P<0.01),and those were significantly lower in JAR-treated HA-SMCs than AngⅡ-induce cells(P<0.05 or P<0.01).The results from RT-PCR showed that expression levels of NOX4 and NOX2 were significantly increased in AngⅡ-induced HA-SMCs compared to those in the control group(P<0.05 or P<0.01),while those were inhibited in JAR-treated HA-SMCs compared to AngⅡ-induced HA-SMCs(P<0.05).Western blot showed that the expression levels of NOX4,NOX2 and p-NF-κB in AngⅡgroup were significantly higher than those in the control group(P<0.01),and JAR intervention could significantly inhibit the expression of the proteins(P<0.05).Conclusion:JAR attenuates AngⅡ-induced HA-SMCs proliferation by inhibiting NOX4/ROS/NF-κB pathway.

关 键 词：熊竹素 平滑肌细胞 增殖 氧化应激 尼克酰胺腺嘌呤二核苷酸磷酸氧化酶4/活性氧/核因子κB 

分 类 号：R965.1[医药卫生—药理学]