主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

北京中医药大学学报 ›› 2015, Vol. 38 ›› Issue (10): 677-681.doi: 10.3969/j.issn.1006-2157.2015.10.007

• 博士之光 • 上一篇    下一篇

左归降糖益肾方药含药血浆对足细胞损伤模型miRNA-30d及GPP78表达的影响

陈聪,曾婧,喻嵘#,刘慧萍,唐元,罗文娟,张翔,米婷   

  1. 湖南中医药大学细胞生物学与分子技术湖南高校重点实验室 湖南 410208
  • 收稿日期:2015-04-12 出版日期:2015-10-30 发布日期:2015-10-30
  • 通讯作者: 喻嵘,女,教授,博士生导师,主要研究方向:内分泌疾病的中医辨证规律及其防治研究,E-mail:yuron@21cn.com
  • 作者简介:陈聪,女,博士
  • 基金资助:
    *国家自然科学基金面上项目(No.81273753),湖南省自然科学基金项目(No.12JJ2048,12JJ9031),湖南省高校创新平台开放基金项目(No.14K069),湖南省研究生科研创新项目(No.CX2013B321),“心脑疾病中西医结合防治研究”湖南省高校科技创新团队资助

Effects of Zuogui Jiangtang Yishen formula medicated serum on the expression of miRNA-30d and GRP78 in podocyte injured model*

CHEN Cong, ZENG Jing, YU Rong#, LIU Huiping, TANG Yuan, LUO Wenjuan, ZHANG Xiang, MI Ting   

  1. Hunan University of Chinese Medicine, Hunan 410208
  • Received:2015-04-12 Online:2015-10-30 Published:2015-10-30

摘要: 目的 探讨左归降糖益肾方药含药血浆处理体外培养的小鼠足细胞对miRNA-30d及葡萄糖调节蛋白78(GRP78)表达水平的影响。方法 将体外培养的小鼠足细胞随机分为空白血浆组(25 mmol/L葡萄糖,10%空白血浆)、模型组(D-葡萄糖50 mmo/L联合棕榈酸2 mmo/L,10%空白血浆)、中药含药血浆组(D-葡萄糖50 mmo/L联合棕榈酸2 mmo/L,10%中药含药血浆),4-苯基丁酸(4-PBA)含药血浆组(D-葡萄糖50 mmo/L联合棕榈酸2 mmo/L,10% 4-PBA含药血浆)。处理48 h后,采用Real-time RT-PCR法检测miRNA-30d表达水平;RT-PCR法、Western blot法检测GRP78 mRNA或蛋白表达水平的变化。结果 与空白组相比,模型组足细胞miRNA-30d表达水平下调、GRP78 mRNA和蛋白的表达水平均上调(P<0.01),且miRNA-30d与GRP78表达水平呈负相关(P<0.05);与模型组相比,中药含药血浆组足细胞miRNA-30d表达水平上调,GRP78 mRNA和蛋白的表达水平均下调,差异均有统计学意义(P<0.01),且在下调miRNA-30d表达水平方面与4-PBA含药血浆组作用相当。结论 左归降糖益肾方药含药血浆可以上调miRNA-30d表达水平,下调GRP78 mRNA和蛋白表达水平,从而改善D-葡萄糖联合棕榈酸诱导的足细胞损伤。

关键词: 足细胞, 左归降糖益肾方药, 含药血浆, miRNA-30d, 葡萄糖调节蛋白78, 小鼠

Abstract: Objective To investigate the expression levels of miRNA-30d and glucose-regulated protein 78kD (GRP78) in podocyte injured model induced by D-glucose (50mmo/L) plus Palmitic acid (2mmol/L), and to observe the changes in miRNA-30d and GRP78 expression levels with Zuogui Jiangtang Yishen (left-restoring sugar-lowering kidney-tonifying) formula in medicated serum 48h after treatment.Methods Cultured podocyte cells were divided into four groups: control group (25mmoL/L glucose, 10% blank serum), model group (D-glucose 50mmo/L combining Palmitic acid 2mmol/L, 10% blank serum), Zuogui Jiangtang Yishen formula medicated serum group (ZJYD, 10% medicated serum, D-glucose 50mmo/L combining Palmitic acid 2mmol/L), and 4-Phenylbutyric acid group (4-PBA, 10% medicated serum, D-glucose 50mmo/L combining Palmitic acid 2mmol/L). After incubation for 48h, the miRNA-30d expression level was analyzed with real-time PCR, and the protein and mRNA expression levels of GRP78 were measured with RT-PCR or western blot respectively. Results The expression level of miRNA-30d was significantly down-regulated in model group, while expression levels of GRP78 protein and mRNA were up-regulated, the differences were statistically significant, as compared with control group (P<0.01), and the expression level of miRNA-30d was inversely correlated with that of GRP78(P<0.05). As compared with model group, the expression level of miRNA-30d was significantly up-regulated in ZJYD group , while down-regulated of GRP78 protein and mRNA(P<0.01), and the effects were partly as good as 4-PBA group. Conclusion 10% ZJYD serum can improve podocyte injury induced by D-glucose 50mmo/L combining Palmitic acid 2mmol/L through up-regulating miRNA-30d expression level, while down-regulating GRP78 mRNA and protein expression levels.

Key words: podocyte, Zuogui Jiangtang Yishen formula, medicated serum, miRNA-30d, glucose-regulated protein 78kD, mouse

中图分类号: 

  • :R285.5