摘要
目的:探讨芪黄补肾泄浊方对高糖培养下人肾小球系膜细胞(HMC)核因子E2相关因子2(Nrf2)/抗氧化反应元件信号通路表达的影响及其对氧化应激的保护作用。方法:离体培养HMC,制备正常大鼠血清,大鼠灌胃给予厄贝沙坦及芪黄补肾泄浊方,达到血药浓度后制备含药血清,实验分为对照组、高糖组、厄贝沙坦组及芪黄补肾泄浊方组。对照组HMC给予正常大鼠血清(10%血清浓度)培养液培养,高糖组HMC予大鼠血清(10%血清浓度)的高糖培养液培养,厄贝沙坦组与芪黄补肾泄浊方组分别给予10%剂量厄贝沙坦与10%剂量芪黄补肾泄浊方含药血清高糖培养液干预培养,培养48 h后收集相关指标检测。采用Real⁃time PCR定量分析的方法检测各组HMC中核因子E2相关因子(Nrf2)、γ⁃谷氨酰半胱氨酸合成酶(γ‐GCS)、超氧化物歧化酶(SOD)的mRNA表达水平。免疫细胞化学法检测各组HMC中γ⁃GCS和SOD的表达;采用Western blot的方法观察各组HMC中γ⁃GCS、SOD蛋白表达的情况。结果:各组实验细胞Nrf2、γ‐GCS、SOD mRNA、蛋白的表达,在对照组中只有少量,高糖组较对照组减少(P<0.05),厄贝沙坦与芪黄补肾泄浊方干预后增加,且以芪黄补肾泄浊方组增加更显著(P<0.05)。结论:芪黄补肾泄浊方可改善高糖培养下HMC氧化应激损伤,其作用机制可能是通过激活Nrf2及其相关下游蛋白γ⁃GCS和SOD达到的。
Objective:To investigate the effect of Qihuang Bushen Xiezhuo Formula on the expression of the nuclear factor E2 related factor 2(Nrf2)/antioxidant response element(ARE)signaling pathway of human glomerular mesangial cells(HMC)in high glucose medium and its protective effect on oxidative stress.Methods:The HMC were cultured in vitro to prepare normal rat serum.The rats were intragastrically administered with irbesartan and Qihuang Bushen Xiezhuo Formula.The serum contain⁃ing the drugs was prepared after the blood concentration was reached.The rats were divided into the control group,the high glu⁃cose group,the irbesartan group and the Qihuang Bushen Xiezhuo Formula group.The HMC of the control group was cultured with normal rat serum(10%serum concentration)medium,while that of the high glucose group was cultured with high glucose medium of rat serum(10%serum concentration).The irbesartan group and the Qihuang Bushen Xiezhuo Formula group were re⁃spectively treated with 10%irbesartan and 10%Qihuang Bushen Xiezhuo Formula in serum high glucose medium.After 48 h of culture,the relevant indicators were collected and detected.The mRNA expression levels of Nrf2,gamma⁃glutamylcysteine syn⁃thetase(γ‐GCS)and superoxide dismutase(SOD)in each group were detected by real⁃time PCR.The expressions ofγ⁃GCS and SOD were detected by immunohistochemistry.The protein expressions ofγ⁃GCS and SOD in HMC of each group were observed by Western Blot.Results:The expressions of Nrf2,γ‐GCS,SOD mRNA and protein in experimental cells of each group were low in the control group,while those in the high glucose group were decreased as compared with the control group(P<0.05).Af⁃ter interference of irbesartan and Qihuang Bushen Xiezhuo Formula the expressions were increased,and the increase in Qihuang Bushen Xiezhuo Formula group was more significant(P<0.05).Conclusion:Qihuang Bushen Xiezhuo Formula can improve HMC oxidative stress injury in high glucose culture,and its mechanism may be achieved by activating Nrf2 and it
作者
金丽霞
张晓东
潘超
栾仲秋
宋淑娟
尹丽颖
王文凯
JIN Li-xia;ZHANG Xiao-dong;PAN Chao;LUAN Zhong-qiu;SONG Shu-juan;YIN Li-ying;WANG Wen-kai(The First Affiliated Hospital of Heilongjiang University of Traditional Chinese Medicine,Harbin 150040,China;Heilongjiang University of Traditional Chinese Medicine,Harbin 150006,China)
出处
《海南医学院学报》
CAS
2022年第20期1533-1539,共7页
Journal of Hainan Medical University
基金
黑龙江省自然科学基金面上项目(H2016066)
黑龙江省中医药学会青年中医药科技创新项目(ZHY19⁃023)。
关键词
芪黄补肾泄浊方
氧化应激
高糖
人肾小球系膜细胞
厄贝沙坦
Qihuang Bushen Xiezhuo Formula
Oxidative stress
High sugar
Human glomerular mesangial cells
Irbesar⁃tan
