摘要
目的:探讨芪黄补肾泄浊方对人肾小球系膜细胞(HMC)核转录因子E2相关因子2(Nrf2)/血红素加氧酶(HO-1)信号通路的影响。方法:体外培养HMC细胞,制备芪黄补肾泄浊方的大鼠含药血清,并用芪黄补肾泄浊方药物血清干预,实验分5组,即正常对照组(A组)、高糖组(B组)、芪黄补肾泄浊方低剂量组(C组)、芪黄补肾泄浊方中剂量组(D组)、芪黄补肾泄浊方高剂量组(E组)。A组HMC给予正常大鼠血清的培养液(血清浓度10%)培养,B组HMC予含正常大鼠血清的高糖培养液(血清浓度10%)培养,C组予含芪黄补肾泄浊方低剂量含药血清的高糖培养液(血清浓度10%)培养干预,D组予含10%芪黄补肾泄浊方中剂量含药血清的高糖培养液(血清浓度10%)培养干预,E组予含10%芪黄补肾泄浊方高剂量含药血清的高糖培养液(血清浓度10%)培养干预,分别于培养48 h末收集细胞进行相关实验指标测定。免疫细胞化学检测方法分析Nrf2及HO-1的表达;运用Western Blot的方法从蛋白质水平观察芪黄补肾泄浊方对HMC Nrf2、HO-1蛋白表达的影响。Real-time PCR定量分析的方法在基因水平探究芪黄补肾泄浊方对HMC中HO-1mRNA表达水平的影响。结果:各组细胞培养48 h末免疫细胞化学法检测结果显示:B组细胞内Nrf2、HO-1的表达显示较A组减少,与B组比较,各药物血清组(C组、D组及E组)细胞内Nrf2、HO-1的表达明显增高,差异具有统计学意义(P<0.05)。Western blot结果显示:与A组比较,Nrf2、HO-1在B组细胞内的表达减少,C组、D组及E组与B组比较,细胞内Nrf2、HO-1的表达明显增高,差异具有统计学意义(P<0.05)。Real-time PCR研究结果显示,各组细胞HO-1mRNA在正常对照组可有少量表达,48 h末高糖组较对照组表达减少,不同剂量芪黄补肾泄浊方药物血清刺激后表达增加,且以中剂量组和高剂量组表达增加更明显(P<0.05)。结论:芪黄补肾泄浊方对高糖培养下HMC氧化应激损伤有改�
Objective: To investigate the effect of Qihuang Bushen Xiezhuo Formula on the signal pathway of nuclear transcription factor E2 related factor 2(Nrf2)/heme oxygenase(HO-1) in human mesangial cells(HMC). Methods: HMC cells were cultured in vitro to prepare the drug-containing serum of Qihuang Bushen Xiezhou Formula. The experiment was divided into five groups, the normal control group(group A), the high sugar group(group B), the low-dose group(Group C), the middle-dose group(Group D) and the high-dose group(Group E). After 48 h corresponding interventions of different concentrations of drug-containing serum, the cells were collected to measure the relevant experimental indicators. Immunocytochemistry was used to analyze the expressions of Nrf2 and HO-1, and Western blot method was used to observe the effect of Qihuang Bushen Xiezhou Formula on the protein expressions of Nrf2 and HO-1 in HMC. Real-time PCR quantitative analysis was used to explore the effect of Qihuang Bushen Xiezhou Formula on the expression of HO-1 mRNA in HMC. Results: In terms of immunocytochemistry results, compared to group A, the expressions of Nrf2 and HO-1 in group B were lower than those in group A;compared to group B, the expressions of Nrf2 and HO-1 in each drug-containing serum group(Group C, D and E) were significantly higher(P<0.05). In terms of Western blot results, compared to group A, the expressions of Nrf2 and HO-1 decreased in group B;the expressions of Nrf2 and HO-1 increased significantly in group C, D and E compared to those in group B(P<0.05). The results of Real-time PCR showed that HO-1 mRNA expressed in a small amount in the normal control group. At the end of 48 hours, the expression of HO-1 mRNA in the high glucose group was lower than that in the control group. After the stimulation of different doses of Qihuang Bushen Xiezhuo Decoction serum, the expression of HO-1 mRNA increased, the increases in the middle dose group and the high dose group were more significant(P<0.05). Conclusion: Qihuang Bushen Xiezhuo Formula c
作者
金丽霞
金丽军
栾仲秋
潘超
马艳春
JIN Lixia;JIN Lijun;LUAN Zhongqiu;PAN Chao;MA Yanchun(The First Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin 150040,China;Mudanjiang Tumor Hospital,Mudanjiang,China;Heilongjiang University of Chinese Medicine,Harbin 150040,China)
出处
《中医药学报》
CAS
2020年第7期21-26,共6页
Acta Chinese Medicine and Pharmacology
基金
黑龙江省第一批省级中医临床优秀人才研修项目
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)。
