Effect of Chaijie Yiganxian on HSC-T6 Cell Proliferation and TGF-/Smad Signaling Pathway

GUO Hui-jun; ZHANG Zhi-bi; YU Xiao-ling; CAI Yue-qing; WEI Wei; ZHANG Rong-ping; JIANG Li-yun

doi:10.13422/j.cnki.syfjx.20181049

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Effect of Chaijie Yiganxian on HSC-T6 Cell Proliferation and TGF-/Smad Signaling Pathway

更新时间：2024-01-04

- Effect of Chaijie Yiganxian on HSC-T6 Cell Proliferation and TGF-/Smad Signaling Pathway
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 13, Pages: 100-104(2018)
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- DOI：10.13422/j.cnki.syfjx.20181049
  CLC： R285.5
- Published：2018
- 稿件说明：
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GUO Hui-jun, ZHANG Zhi-bi, YU Xiao-ling, et al. Effect of Chaijie Yiganxian on HSC-T6 Cell Proliferation and TGF-/Smad Signaling Pathway[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(13): 100-104.
DOI：

GUO Hui-jun, ZHANG Zhi-bi, YU Xiao-ling, et al. Effect of Chaijie Yiganxian on HSC-T6 Cell Proliferation and TGF-/Smad Signaling Pathway[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(13): 100-104. DOI： 10.13422/j.cnki.syfjx.20181049.

摘要

目的：研究保肝护肝中药复方柴竭抑肝纤（CJYGX）通过肝星状细胞治疗肝纤维化的作用机制。方法：培养肝星状细胞T6（hepatic stellate cell T6，HSC-T6），设空白组、秋水仙碱1.0 mg·L-1组和200，100，50，25，12.5，6.25 mg·L-1质量CJYGX组，作用24 h后通过细胞计数试剂（cell counting kit-8，CCK-8）检测HSC-T6细胞的增殖情况，筛选合适的CJYGX药物浓度；实时荧光定量聚合酶链式反应（Real-time PCR）检测CJYGX（200，100，50 mg·L-1）作用HSC-T6细胞24 h后转化生长因子-β1（TGF-β1），信号转导蛋白Smad2，Smad3，Smad4，Smad7 mRNA表达变化；蛋白免疫印迹法（Western blot）检测CJYGX（200，100，50 mg·L-1）作用HSC-T6细胞24 h后TGF-β1，α-平滑肌肌动蛋白（α-SMA）表达变化。结果：①与空白组比较，200，100，50 mg·L-1CJYGX对HSC-T6细胞具有显著的抑制增殖作用（P<0.05）。②200，100，50 mg·L-1CJYGX显著下调TGF-β1，Smad2，Smad3 mRNA的表达（P<0.05）；200，100 mg·L-1CJYGX显著下调Smad4 mRNA表达，并显著上调Smad7 mRNA表达（P<0.05）。③ 200，100 mg·L-1CJYGX显著下调HSC-T6细胞TGF-β1和α-SMA蛋白表达（P<0.05）。结论：CJYGX能抑制HSC-T6细胞的增殖，其作用机制与TGF-β1/Smad通路有关。

Abstract

Objective: To study the protective mechanism of Chaijie Yiganxian (CJYGX) on the liver fibrosis through hepatic stellate cells (HSC-T6) and transforming growth factor-β1/signal transduction protein (TGF-β1/Smad) signaling pathway. Method: Different concentrations of colchicine (1.0 mg·L-1) and CJYGX (200

100

12.5

6.25 mg·L-1) were added to the culture medium

and the proliferation of HSC-T6 was detected by cell counting kit-8 (CCK-8) after 24 hours. The appropriate drug concentration that can inhibit the cell proliferation was screened out according to the CCK-8 results. The mRNA expressions of TGF-β1

Smad2

Smad3

Smad4 and Smad7 in HSC-T6 cells treated with CJYGX (200

100

50 mg·L-1) were detected by Real-time PCR

and the protein expressions of TGF-β1 and α-SMA also were detected by Western blot. Result: Compared with the blank group

CJYGX (200

100

50 mg·L-1) had a significantly inhibitory effect on HSC-T6 cells (P<0.05).CJYGX (200

100

50 mg·L-1) can significantly down-regulate the mRNA expressions of TGF-β1

Smad2 and Smad3 (P<0.05). Meanwhile

CJYGX (200

100 mg·L-1) can significantly down-regulate Smad4 mRNA expression and up-regulated Smad7 mRNA expression (P<0.05).CJYGX (200

100 mg·L-1) can significantly down-regulate the protein expressions of TGF-β1 and α-SMA in HSC-T6 cells(P<0.05). Conclusion: CJYGX can inhibit HSC-T6 cell proliferation

and its mechanism is related to TGF-β1/Smad signaling pathway.

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