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
高俊丽 1张骢翀 1谷亮 2周建 1费秀丽 1姚卫国 1周锦明 1刘琨1
作者信息
- 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)
- 折叠
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1g/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-β1,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-β1、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-β1、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-β1 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.
Abstract
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 op-timal 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 proteins α-SMA,FN,TGF-β1,mitochondrial fis-sion protein Drp1 and mitochondrial fusion protein mfn2,and transmission electron microscopy was used to observe mitochondrial mor-phology.Results:(1)Compared with the normal group,the mRNA and protein expressions of α-SMA,FN,TGF-β1 and Drp1 in the high glucose group were significantly increased(P<0.001);The mRNA and protein expressions of Mfn2 were significantly de-creased(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 of α-SMA,FN,TGF-β1 and Drp1 in the Huangkui group were significantly decreased(P<0.01),and the mRNA and protein ex-pressions 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 frag-ments,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 the production of α-SMA,FN and TGF-β1,and inhibit HK2 fibrosis induced by high glucose.At the same time,it can promote the expression of mitochondrial fu-sion protein Mfn2 and reduce the expression of mitochondrial fission protein Drp1,thereby reversing renal fibrosis and delaying the progression of diabetic nephropathy.
关键词
HK2/黄葵浸膏粉/高糖/纤维化/线粒体分裂及融合Key words
HK2/Huangkui extract powder/High glucose/Fibrosis/Mitochondrial fission and fusion引用本文复制引用
基金项目
上海市金山区科学技术委员会项目(2019-3-9)
上海市卫生健康委员会科研项目(20214Y0144)
周锦明上海市名老中医学术经验研究工作室(SHGZS-202237)
上海市金山区第七周期医学重点专科建设项目(JSZK2023H04)
上海市"十四五"中医特色专科建设项目(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(JSYQ202310)
出版年
2024
NETL
NSTL
万方数据