摘要
目的 观察槐杞黄颗粒含药血清(以下简称HQH)对高糖培养的肾小球系膜细胞增殖的影响,并探讨其可能机制。方法 用HQH及生理盐水分别对SPF级雄性SD大鼠进行灌胃,制备HQH含药血清与正常血清。将HBZY-1细胞分为正常糖组,高糖组,高糖+低、中、高浓度HQH含药血清组(HQH含药血清浓度分别为5%、10%、15%),CCK-8法检测各组细胞增殖率,筛选出有效浓度HQH含药血清用于后续实验;倒置光学显微镜下观察各组HBZY-1细胞形态并拍照;流式细胞仪检测各组细胞凋亡率;试剂盒检测各组细胞超氧化物歧化酶(Superoxide dismutase,SOD)、过氧化氢酶(Catalase,CAT)、谷胱甘肽过氧化物酶(Glutathione peroxidase,GSH-Px)和丙二醛(Malondialdehyde,MDA)活性;Western blot检测各组细胞TLR4/NF-κB/NLRP3炎症通路相关蛋白的表达。结果 与高糖组相比,5%含药血清组细胞增殖率变化不明显,10%含药血清组和15%含药血清组细胞增殖率明显下降(P<0.05或P<0.01),因此,选择10%和15%浓度HQH含药血清作为后续实验用药处理组。对高糖环境下的HBZY-1细胞予以10%含药血清和15%含药血清干预后,与高糖组相比,10%含药血清组和15%含药血清组细胞增殖度降低,细胞凋亡比例逐渐增加,细胞SOD、CAT、GSH-Px活性升高(P<0.05或P<0.01),MDA水平降低(P<0.01),TLR4、NLRP3、p-NF-κB p65和p-IκBα蛋白表达明显下降(P<0.01)。结论 槐杞黄颗粒含药血清能够抑制高糖环境下HBZY-1细胞增殖并诱导其凋亡,其机制可能与抑制细胞氧化应激及TLR4/NF-κB/NLRP3介导的炎症信号通路有关。
Objective To observe the effect of Huaiqihuang granules(HQH)containing serum on the proliferation in rat glomerular mesangial cells(HBZY-1)cultured with high glucose,and to explore its possible mechanism.Methods HQH and physiological saline were administered orally to SPF grade male SD rats for preparing HQH containing serum and normal serum.HBZY-1 cells were divided into normal glucose group,high glucose group,high glucose+low,medium,and high concentration HQH drug containing serum group(HQH drug containing serum concentration was 5%,10%,and 15%,respectively).The cell proliferation rate of each group was measured by CCK-8 method,and effective HQH drug containing serum was screened for subsequent experiments;Observe the morphology of HBZY-1 cells in each group using an inverted optical microscope and take photos;Flow cytometry was used to detect the apoptosis rate;The assay kit was used to detect the activities of superoxide dismutase(SOD),catalase(CAT),glutathione peroxidase(GSH-Px),and malondialdehyde(MDA)in each group of cells;Western blot was used to detect the expression of TLR4/NF-κB/NLRP3 inflammatory pathway related proteins in each group of cells.Results Compared with the high glucose group,the 5%medicated serum group showed no significant change in cell proliferation rate,while the 10%medicated serum group and 15%medicated serum group showed a significant decrease in cell proliferation rate(P<0.05 or P<0.01).Therefore,10%and 15%concentrations of HQH medicated serum were selected as the subsequent experimental drug treatment groups.After intervention with 10%medicated serum and 15%medicated serum on HBZY-1 cells in a high glucose environment,compared with the high glucose group,the 10%medicated serum group and 15%medicated serum group showed a decrease in cell proliferation,a gradual increase in cell apoptosis,an increase in cell SOD,CAT,GSH-Px activity(P<0.05 or P<0.01),a decrease in MDA levels(P<0.01),the protein expression of TLR4,NLRP3,pNF-κB p65 and p-IκBαsignificantly decreased(P<0.01).Conclusion Huaiqihuang granules containing serum can inhibit high glucose induced proliferation and induce apoptosis of HBZY-1 cells,which may be related to the inhibition of cellular oxidative stress and TLR4/NF-κB/NLRP3 mediated inflammatory signaling pathway.
作者
刘红
姜南
邱文慧
薛雪
LIU Hong;JIANG Nan;QIU Wenhui;XUE Xue(Wuhan No.1 Hospital,Wuhan 430022,China;Affiliated Hospital of Hubei University of Chinese Medicine,Hubei Shizhen Laboratory,Hubei Key Laboratory of Theory and Application Research of Liver and kidney in Traditional Chinese Medicine,Wuhan 430061,China)
出处
《世界科学技术-中医药现代化》
北大核心
2025年第3期815-822,共8页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
武汉市卫生健康委员会中医药科研青年项目(WZ22Q36):槐杞黄颗粒通过抑制氧化应激和TLR4/NF-κB/NLRP3炎症信号通路延缓糖肾进展的实验研究,负责人:刘红
湖北省自然科学基金项目(2022CFC056):基于AMPK通路探讨IgA血管炎肾炎Treg/Th17失衡的机制及益气养阴化瘀通络方药的干预作用,负责人:薛雪。
