黄葵浸膏粉对高糖诱导的人肾小管上皮细胞纤维化的影响  

Effect of Yellow Sunflower Extract Powder on Fibrosis of Human Renal Tubular Epithelial Cells Induced by High Glucose

作　　者：高俊丽 张骢翀[1] 谷亮 周建 费秀丽 姚卫国 周锦明[1] 刘琨[1] GAO Junli;ZHANG Congchong;GU Liang(Jinshan Branch,Shanghai Sixth People’s Hspital,Shanghai,201500)

机构地区：[1]上海市第六人民医院金山分院肾内科,上海201500 [2]复旦大学附属中山医院青浦分院呼吸内科,上海201700

出　　处：《中国中西医结合肾病杂志》2024年第7期578-581,I0002,I0003,共6页Chinese Journal of Integrated Traditional and Western Nephrology

基　　金：上海市金山区科学技术委员会项目(No.2019-3-9);上海市卫生健康委员会科研项目(No.20214Y0144);周锦明上海市名老中医学术经验研究工作室(No.SHGZS-202237);上海市金山区第七周期医学重点专科建设项目(No.JSZK2023H04);上海市“十四五”中医特色专科建设项目(No.ZYTSZK2-13);上海市金山区卫生系统第五周期“优秀青年人才”培养计划项目(No.JSYQ202310)。

摘　　要：目的:研究黄葵浸膏粉对高糖诱导的人肾小管上皮细胞(HK2)纤维化影响,探讨其改善肾纤维化的可能机制。方法:不同浓度黄葵浸膏粉干预24 h、48 h、72 h,CCK8检测细胞增殖,最佳干预浓度1 g/L、时间48 h。分组:正常组(葡萄糖5.5 mmol/L)、甘露醇组(葡萄糖5.5 mmol/L+甘露醇24.5 mmol/L)、高糖组(葡萄糖30 mmol/L)及黄葵组(葡萄糖30 mmol/L+黄葵浸膏粉1 g/L),干预48 h,qPCR及Western Blot检测纤维化相关蛋白α-SMA、FN、TGF-β_(1),线粒体分裂蛋白Drp1及线粒体融合蛋白mfn2表达,透射电镜观察线粒体形态。结果:(1)与正常组比,高糖组α-SMA、FN、TGF-β_(1)、Drp1的mRNA和蛋白表达显著升高(P<0.001);Mfn2的mRNA和蛋白表达显著降低(P<0.001)。(2)与正常组比,甘露醇组指标差异均无统计学意义(P>0.05),即高浓度渗透压对HK2无作用。(3)与高糖组比,黄葵组α-SMA、FN、TGF-β_(1)、Drp1的mRNA和蛋白表达显著降低(P<0.01),Mfn2的mRNA和蛋白表达显著升高(P<0.01)。(4)透射电镜:正常组及甘露醇组线粒体数量较多,形态正常;高糖组可见肿胀的线粒体,线粒体碎片较多,线粒体基质及线粒体嵴断裂;黄葵浸膏粉干预后可减轻线粒体损伤。结论:黄葵浸膏粉可减少α-SMA、FN、TGF-β_(1)的产生,抑制高糖诱导的HK2纤维化;同时可促进线粒体融合蛋白Mfn2表达,减少线粒体分裂蛋白Drp1表达,从而逆转肾纤维化,延缓糖尿病肾病进展。Objective:To investigate the effect of Huangkui extract powder on the fibrosis of human renal tubular epithelial cells(HK2)induced by high glucose,and to explore the possible mechanism of improving renal fibrosis.Methods:CCK8 was used to detect cell proliferation after 24 h,48 h and 72 h intervention with different concentrations of Huangkui extract powder,and the optimal intervention concentration was 1 g/L and 48 h.Grouping:Normal group(glucose 5.5 mmol/L),mannitol group(glucose 5.5 mmol/L+mannitol 24.5 mmol/L),high glucose group(glucose 30 mmol/L)and yellow sunflower group(glucose 30 mmol/L+yellow sunflower extract powder 1 g/L),intervention for 48 hours,and mannitol group(glucose 5.5 mmol/L+mannitol 24.5 mmol/L).qPCR and Western Blot were used to detect the expressions of fibrosis-related proteinsα-SMA,FN,TGF-β_(1),mitochondrial fission protein Drp1 and mitochondrial fusion protein mfn2,and transmission electron microscopy was used to observe mitochondrial morphology.Results:(1)Compared with the normal group,the mRNA and protein expressions ofα-SMA,FN,TGF-β_(1) and Drp1 in the high glucose group were significantly increased(P<0.001);The mRNA and protein expressions of Mfn2 were significantly decreased(P<0.001).(2)Compared with the normal group,there was no difference in the indexes of the mannitol group(P>0.05),that is,high osmolality had no effect on HK2.(3)Compared with the high glucose group,the mRNA and protein expressions ofα-SMA,FN,TGF-β_(1) and Drp1 in the Huangkui group were significantly decreased(P<0.01),and the mRNA and protein expressions of Mfn2 were significantly increased(P<0.01).(4)Transmission electron microscopy:the number of mitochondria in the normal group and the mannitol group was large,and the morphology was normal.Swollen mitochondria,more mitochondrial fragments,mitochondrial matrix and mitochondrial cristae disruption were observed in the high glucose group.Huangkui extract powder can reduce mitochondrial damage after intervention.Conclusion:Huangkui extract powder can reduce t

关 键 词：HK2 黄葵浸膏粉 高糖 纤维化 线粒体分裂及融合 

分 类 号：R285[医药卫生—中药学]