Effect of Sodium Ferulate on Proliferation, Activation and Collagen Synthesis of Hepatic Stellate Cells Induced by Acetaldehyde

ZHANG Zhi-bi; CAI Ting; MIAO Zi-e; XU Zhe; FENG Guo-hua

doi:10.13422/j.cnki.syfjx.20181132

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Effect of Sodium Ferulate on Proliferation, Activation and Collagen Synthesis of Hepatic Stellate Cells Induced by Acetaldehyde

更新时间：2024-01-04

- Effect of Sodium Ferulate on Proliferation, Activation and Collagen Synthesis of Hepatic Stellate Cells Induced by Acetaldehyde
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 12, Pages: 123-128(2018)
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- DOI：10.13422/j.cnki.syfjx.20181132
  CLC： R285
- Published：2018
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ZHANG Zhi-bi, CAI Ting, MIAO Zi-e, et al. Effect of Sodium Ferulate on Proliferation, Activation and Collagen Synthesis of Hepatic Stellate Cells Induced by Acetaldehyde[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(12): 123-128.
DOI：

ZHANG Zhi-bi, CAI Ting, MIAO Zi-e, et al. Effect of Sodium Ferulate on Proliferation, Activation and Collagen Synthesis of Hepatic Stellate Cells Induced by Acetaldehyde[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(12): 123-128. DOI： 10.13422/j.cnki.syfjx.20181132.

摘要

目的：研究阿魏酸钠（SF）对体外乙醛诱导的大鼠肝星状细胞（HSC-T6）增殖、活化、胶原合成的影响及其作用机制。方法：体外培养HSC-T6细胞，建立乙醛诱导的HSC-T6增殖模型，设空白组、乙醛组、秋水仙碱组（2.5 μmol·L-1）及SF组（1，10，25，50，100，200，400 μmol·L-1），通过细胞增殖和毒性检测（MTS）比色法检测细胞增殖，筛选合适的SF浓度。蛋白免疫印迹法（Western blot）检测SF （400，200，100 μmol·L-1）对乙醛诱导的HSC-T6细胞α-平滑肌肌动蛋白（α-SMA）表达；酶联免疫吸附测定法（ELISA）检测Ⅰ型，Ⅲ型胶原浓度，酸水解法检测羟脯氨酸（Hyp）含量，实时荧光定量聚合酶链式反应（Real-time PCR）检测转化生长因子-β1/（TGF-β1/），信号转导蛋白Smad2，Smad3，Smad4，Smad7，基质金属蛋白酶-1（MMP-1）和金属蛋白酶抑制剂-1（TIMP-1）基因mRNA表达变化。结果：与空白组比较，200 μmol·L-1的乙醛能显著诱导HSC-T6体外增殖（P<0.01）；与空白组比较，模型组HSC-T6增殖显著增加（P<0.01），α-SMA和I型，Ⅲ型胶原和Hyp含量显著增加（P<0.01），TGF-β1/，Smad2，Smad3，Smad4，Smad7 mRNA的表达显著上调（P<0.01），MMP-1和TIMP-1 mRNA表达显著上调（P<0.01）。与模型组比较，400，200，100 μmol·L-1浓度的SF能明显抑制乙醛诱导的HSC-T6增殖（P<0.05，P<0.01），显著降低α-SMA和I型，Ⅲ型胶原和Hyp含量（P<0.01），下调TGF-β1/，Smad2，Smad3，Smad4基因mRNA的表达（P<0.05，P<0.01），上调Smad7基因mRNA表达（P<0.01），并明显上调MMP-1 mRNA表达（P<0.05，P<0.01）和下调TIMP-1 mRNA表达（P<0.01）。结论： SF能通过调控TGF-β1/Smad和MMP-1/TIMP-1信号通路，抑制乙醛诱导的体外HSC-T6细胞的增殖，活化和胶原分泌。

Abstract

Objective: To investigate the effect of sodium ferulate (SF) on the proliferation

activation and collagen synthesis of rat hepatic stellate cells (HSC-T6) induced by acetaldehyde in vitro and explore its mechanism. Method: HSC-T6 cells were cultured in vitro to establish acetaldehyde-induced HSC-T6 proliferation models. The rats were divided into blank group

acetaldehyde group

colchicine group (2.5 μmol·L-1)

and SF groups (1

100

200

400 μmol·L-1). HSC-T6 proliferation was measured by MTS method and the appropriate SF concentration was screened. The effect of SF (400

200

100 μmol·L-1) on the expression of α-smooth muscle actin (α-SMA) in HSC-T6 cells induced by acetaldehyde was detected by Western blot; type I and type Ⅲ collagen concentrations were detected by Enzyme-linked immunosorbent assay (ELISA); the Hydroxyproline (Hyp) content was detected by hydrolysis method; mRNA expression levels of transforming growth factor-β1 (TGF-β1)

signal transducers Smad2

Smad3

Smad4

Smad7

Matrix metallo proteinase 1 (MMP-1) and tissue inhibitors of metallo proteinase-1 (TIMP-1) were detected by real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). Result: As compared with the blank group

200 μmol·L-1 acetaldehyde significantly induced HSC-T6 proliferation in vitro (P<0.01). As compared with the blank group

the proliferation of HSC-T6 in model group was significantly increased (P<0.01)

and the contents of α-SMA

type I

type Ⅲ collagen and Hyp were significantly increased (P<0.01)

the mRNA expression levels of TGF-β1

Smad2

Smad3

Smad4 and Smad7 as well as mRNA expression levels of MMP-1 and TIMP-1 were significantly up-regulated (P<0.01). As compared with the model group

SF at concentrations of 400

200

100 μmol·L-1 significantly inhibited acetaldehyde-induced HSC-T6 proliferation (P<0.05

P<0.01)

significantly decreased α-SMA

type I and type Ⅲ collagen and Hyp contents (P<0.05

P<0.01)

down-regulated the mRNA expression of TGF-β1

Smad2

Smad3 and Smad4 (P <0.05

P<0.01)

up-regulated the mRNA expression of Smad7 mRNA (P<0.05

P<0.01) and down-regulated the mRNA expression of TIMP-1 (P<0.01). Conclusion: SF can inhibit proliferation

activation and collagen secretion of HSC-T6 induced by acetaldehyde in vitro by regulating TGF-β1/Smads and MMP-1/TIMP-1 signaling pathways.

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Related Institution

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