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 ›› 2021, Vol. 44 ›› Issue (5): 421-427.doi: 10.3969/j.issn.1006-2157.2021.05.006

• 中药药理 • 上一篇    下一篇

甘草酸对TGF-β1诱导的NRK-49F细胞活化及分泌细胞外基质的影响*

邸黎明, 张澜, 申萌萌, 贺双双, 李彧#   

  1. 北京中医药大学中医学院 北京 100029
  • 收稿日期:2020-10-31 出版日期:2021-05-30 发布日期:2021-06-01
  • 通讯作者: #李彧,女,博士,研究员,博士生导师,主要研究方向:中医药防治多器官纤维化,E-mail:liyubeijing1973@163.com
  • 作者简介:邸黎明,女,在读硕士生
  • 基金资助:
    *国家自然科学基金资助项目(No.81573716),北京中医药大学科研创新团队项目(No.2019-JYB-TD-006)

Effect of glycyrrhizic acid on the activation and extracellular matrix secretion in renal interstitial fibroblast cells induced by transforming growth factor β1*

Di Liming, Zhang Lan, Shen Mengmeng, He Shuangshuang, Li Yu#   

  1. School of Chinese Medicine, Beijing University of Chinese Medicine, Beijing 100029, China
  • Received:2020-10-31 Online:2021-05-30 Published:2021-06-01
  • Contact: Li Yu, Ph. D., Researcher, Doctoral Supervisor. School of Chinese Medicine, Beijing University of Chinese Medicine, No. 11, Beisanhuan East Road, Chaoyang District, Beijing 100029. E-mail: liyubeijing1973@163com
  • Supported by:
    National Natural Science Foundation of China(No.81573716), Innovation Team of Beijing University of Chinese Medicine (No.2019-JYB-TD-006)

摘要: 目的 探讨甘草酸对转化生长因子β1(TGF-β1)诱导的大鼠肾间质成纤维细胞(NRK-49F)中纤维化相关因子表达的影响。方法 体外培养NRK-49F细胞,不同浓度甘草酸干预细胞48 h后,CCK-8法检测其对细胞增殖的影响,确定对细胞活性无显著影响的作用浓度。以10 μg/L TGF-β1刺激NRK-49F细胞构建肾间质纤维化细胞模型,加入不同浓度甘草酸进行干预,免疫荧光法检测α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原(COL1)表达情况。实时荧光定量PCR、Western Blot法检测纤维化相关因子α-SMA、COL1、Ⅲ型胶原(COL3) mRNA及蛋白表达情况,以评价甘草酸对成纤维细胞活化及细胞外基质分泌的影响。结果 CCK-8结果显示甘草酸在100~600 μmol/L浓度范围内对NRK-49F细胞活性无影响。免疫荧光结果表明,加入甘草酸后可抑制α-SMA表达及COL1分泌,随甘草酸浓度增加,抑制作用增强。Western Blot结果显示,与模型组相比,甘草酸干预可下调α-SMA、COL1、COL3的蛋白表达,其中甘草酸150 μmol/L组COL1的蛋白表达水平降低(P<0.01),甘草酸200 μmol/L组COL1、COL3表达均降低(P<0.01)。RT-PCR结果表明,与模型组相比,甘草酸干预可下调α-SMA、COL1、COL3表达,甘草酸150 μmol/L组COL1 mRNA的表达水平降低(P<0.01),甘草酸200 μmol/L组COL1、COL3 mRNA表达均降低(P<0.01)。结论 甘草酸能够有效抑制TGF-β1诱导的NRK-49F细胞中α-SMA、COL1、COL3的mRNA及蛋白表达,其作用机制有待进一步探讨。

关键词: 甘草酸, 转化生长因子β1, Ⅰ型胶原, Ⅲ型胶原, α-平滑肌肌动蛋白, 大鼠肾间质成纤维细胞

Abstract: Objective To investigate the effect of glycyrrhizic acid (GA) on the expression of fibrosis-related factors in the renal interstitial fibroblastcells (NRK49F) of rats induced by transforming growth factor β1(TGF-β1).Methods NRK-49F cells were cultured in vitro.After 48 h of intervention with different concentrations of GA,Cell Counting Kit-8 was used to detect their effect on cell proliferation, so as to determine the concentration with no obvious effect on cell viability.The model was established by stimulating NRK-49F cells with 10 μg/L TGF-β1. Then different concentrations of GA were used for intervention. The expression of α-SMA and type 1 collagen (COL1) was detected with immunofluorescence method. The mRNA and protein expression of fibrosis-related factors of α-SMA,COL1, COL3 was detected with RT-PCR and WB methods, so as to evaluate the effect of GA on the activation of fibroblasts and the secretion of extracellular matrix.Results CCK-8 results showed that within the concentration range of 100~600 μmol/L, GA had no effect on the activity of NRK-49F cells.According to immunofluorescence results, the addition of GA intervention inhibited the expression of α-SMA and the secretion of COL1, with the increase of GA concentration,the inhibitory effect being enhanced. According to Western Blot results, GA intervention could down-regulate the expression of α-SMA, COL1 and COL3 compared with the model group; the expression level of COL1 protein in the GA 150 μmol/L group was reduced (P<0.01), and the level of COL1 and COL3 in the GA 200 μmol/L group was reduced (P<0.01). RT-PCR results showed that GA intervention could down-regulate the expression of α-SMA, COL1 and COL3 compared with the model group; among them, the mRNA expression level of COL1 in the GA 150 μmol/L group was reduced (P<0.01), and the expression of COL1 and COL3 in the GA 200 μmol/L group was reduced (P<0.01).Conclusion GA seems to inhibit the mRNA and protein expression of α-SMA, COL1 and COL3 in NRK-49F cells induced by TGF-β1, but its mechanism still needs to be further explored.

Key words: glycyrrhizic acid, TGF-β1, COL1, COL3, α-SMA, NRK-49F

中图分类号: 

  • R285.5