摘要
目的探讨益气通络汤对体外膜攻击复合物(C5b-9)诱导的小鼠肾足细胞损伤模型的干预作用,以期明确益气通络汤在膜性肾病治疗中的作用。方法将小鼠足细胞(MPC5)完全随机分为空白对照组,模型组,低、中、高剂量含药血清组,厄贝沙坦组。以C5b-9(0.8μg/mL)诱导培养的MPC5细胞建立足细胞损伤模型。CCK-8法检测益气通络汤对损伤足细胞活性的影响;微板法测定乳酸脱氢酶(LDH)活性,检测益气通络汤对足细胞损伤的影响;流式细胞术检测益气通络汤对足细胞凋亡的影响;双抗体夹心法(ELISA法)测定白细胞介素1β(IL-1β)水平;流式细胞仪检测活性氧(ROS)荧光强度,检测益气通络汤对足细胞氧化应激的影响。结果低、中、高剂量含药血清组和厄贝沙坦组足细胞OD值均大于模型组,差异均有统计学意义(P<0.05)。低、中、高剂量含药血清组和厄贝沙坦组LDH水平均低于模型组,差异均有统计学意义(P<0.05)。低、中、高剂量含药血清组和厄贝沙坦组足细胞晚期细胞凋亡百分比均低于模型组,差异均有统计学意义(P<0.05)。正常的活细胞不被Annexin V-FITC和碘化丙啶(PI)染色;凋亡早期的细胞仅被Annexin V-FITC染色,PI染色呈阴性;凋亡晚期的细胞可以同时被Annexin V-FITC和PI染色;坏死细胞不能被Annexin V-FITC染色,只能被PI染色。低、中、高剂量含药血清组和厄贝沙坦组足细胞炎症因子IL-1β的OD值均小于模型组,差异均有统计学意义(P<0.05)。低、中、高剂量含药血清组和厄贝沙坦组足细胞ROS荧光强度均低于模型组,差异均有统计学意义(P<0.05)。结论益气通络汤可以减轻足细胞损伤,对足细胞活性有正向诱导作用,且能够提高足细胞的抗氧化能力,其可能是通过抑制足细胞的氧化应激、细胞凋亡和炎症反应等途径实现的。
Objective This paper aims to investigate the effect of Yiqi Tongluo Decoction on renal podocyte injury induced by extracorporeal complement membrane attack complex(C5b-9)in mice and to clarify its effect in the treatment of membranous nephropathy.Methods Mouse podocyte clone 5(MPC5)was randomly assigned to a blank control group,a model group,low,medium-and high-dose medicated serum groups,and an irbesartan group.A podocyte injury model was established by treating cultured MPC5 with C5b-9(0.8μg/mL).CCK-8 assay was used to detect the effect of Yiqi Tongluo Decoction on the activity of injured podocytes;lactate dehydrogenase(LDH)activity was measured by microplate assay to detect its effect on podocyte injury;flow cytometry was used to detect its effect on podocyte apoptosis;double-antibody sandwich assay(ELISA)was used to determine interleukin 1β(IL-1β)levels;reactive oxygen species(ROS)fluorescence intensity was measured by flow cytometry to detect its effect on oxidative stress in podocytes.Results The OD values of podocytes in low,medium,and high-dose medicated serum groups and the irbesartan group were significantly higher than those in the model group(P<0.05).LDH levels in the low,medium,and high-dose medicated serum groups and the irbesartan group were significantly lower than those in the model group(P<0.05).The percentages of late podocyte apoptosis in the low,medium,and high-dose medicated serum groups and irbesartan group were significantly lower than those in the model group(P<0.05).normal viable cells are not stained by Annexin V-FITC and propidium iodide(PI);cells in the early stage of apoptosis only stained by Annexin V-FITC and stained negatively by propidium iodide;cells in the late stage of apoptosis can be stained by Annexin V-FITC and PI simultaneously;necrotic cells cannot be stained by Annexin V-FITC and can only be stained by PI.The OD values of IL-1β,an inflammatory factor in podocytes,in the low,medium,and high-dose medicated serum groups and irbesartan group were significantly lower than those in the model group(P<0.05).ROS fluorescence intensity in podocytes was significantly lower in the low,medium,and high-dose medicated serum groups and irbesartan group than in the model group(P<0.05).Conclusion Yiqi Tongluo Decoction can reduce podocyte injury,positively induce podocyte activity,and improve the antioxidant capacity of podocytes.The mechanism may be through inhibiting oxidative stress,apoptosis,and inflammatory response pathways of podocytes.
作者
甘露
杨震
张春戬
李莹
郝芳艺
董壮
郑佳新
Ganlu;Yang Zhen;Zhang Chunjian;Li Ying;Hao Fangyi;Dong Zhuang;Zheng Jiaxin(Heilongjiang Academy of Chinese Medical Sciences,Harbin 150036,China;Department of Nephrology,The Second Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin 150001,China)
出处
《保健医学研究与实践》
2023年第9期1-7,F0002,共8页
Health Medicine Research and Practice
基金
黑龙江省重点研发计划项目(2022ZX06C16)
黑龙江省中医药科研项目(ZHY2020-049)。
关键词
益气通络汤
足细胞损伤
细胞凋亡
炎症因子
Yiqi Tongluo Decoction
Podocyte injury
Apoptosis
Inflammatory factors
