姜黄素及其衍生物对TGF-β诱导的LX-2细胞纤维化的抑制作用研究

邵益丹; 史婷婷; 赵艳梅; 邹玺; 席建军; 张晶; 姜晓杰; 庄让笑

doi:10.13748/j.cnki.issn1007-7693.20233169

姜黄素及其衍生物对TGF-β诱导的LX-2细胞纤维化的抑制作用研究

Inhibitory Effect of Curcumin and Its Derivatives on TGF-β Induced Fibrosis of LX-2 Cells

摘要

摘要:
目的研究姜黄素及其衍生物A和B对TGF-β诱导的LX-2细胞纤维化的抑制作用及其机制。
方法采用TGF-β(10 ng·mL⁻¹ )诱导LX-2细胞肝纤维化模型；CCK-8法检测姜黄素及其衍生物对细胞增殖的影响；流式细胞术检测细胞凋亡率；Western blotting和q-PCR法检测纤维化相关因子Collagen Ⅰ、Collagen Ⅳ、Fibronectin、Vimentin、α-SMA、PDGFRβ、TGFβR1、TGFβR2、MMP2、MMP9、TIMP1和TIMP2蛋白表达和基因转录水平。
结果姜黄素及其衍生物A和B对正常LX-2细胞的生长具有抑制作用，IC₂₅值分别为15.7、2.6和10.2 μmol·L⁻¹；与模型组比较，姜黄素(15.7 μmol·L⁻¹)及其衍生物A(2.6 μmol·L⁻¹)和B(10.2 μmol·L⁻¹)对TGF-β诱导LX-2细胞增殖具有抑制作用(P<0.05)；姜黄素衍生物B组的细胞凋亡率升高；姜黄素组的Collagen I、Fibronectin、Vimentin、α-SMA、TGFβR1和TIMP-1的蛋白表达水平下降，MMP-9的蛋白表达水平升高(P<0.05)；姜黄素衍生物A组的Collagen I、Collagen IV、Fibronectin、Vimentin、α-SMA、TIMP-1、TIMP-2的蛋白表达水平下降，MMP-2的蛋白表达水平升高(P<0.05)；姜黄素衍生物B组的Collagen I、Collagen IV、Fibronectin、Vimentin、α-SMA、PDGFRβ、TGFβR1、TGFβR2、TIMP-1、TIMP-2的蛋白表达水平下降，MMP-2的蛋白表达水平升高(P<0.05)。姜黄素组的Collagen I、Fibronectin、α-SMA、TIMP-1的基因转录水平下降(P<0.05)。姜黄素衍生物A组和B组的Collagen I、Fibronectin、α-SMA的基因转录水平下降(P<0.05)。
结论姜黄素及其衍生物A和B通过抑制TGF-β诱导的LX-2细胞的异常活化和增殖，抑制其细胞外基质组分的过度分泌和积聚，促进其降解，从而发挥体外抗纤维化作用，尤以姜黄素衍生物B的作用最突出。

Abstract:
OBJECTIVE To study the inhibitory effect and mechanism of curcumin and its derivatives A and B on TGF-β induced LX-2 cell fibrosis.
METHODS Established the liver fibrosis model of LX-2 cells induced by TGF-β(10 ng·mL⁻¹).The effects on cell proliferation were detected by CCK-8. The effects on cell apoptosis was detected by flow cytometry. The effects on fibrosis related factors(Collagen I, Collagen Ⅳ, Fibronectin, Vimentin, α-SMA, PDGFRβ, TGFβR1, TGFβR2, MMP2, MMP9, TIMP1 and TIMP2) protein expression and gene transcription levels were detected by Western blotting and q-PCR.
RESULTS The curcumin and its derivative A and B had the inhibition effects on normal LX-2 cells, and the IC₂₅ values were 15.7, 2.6, 10.2 μmol·L⁻¹, respectively. Compared to the model group, the curcumin(15.7 μmol·L⁻¹) and its derivative A(2.6 μmol·L⁻¹) and B(10.2 μmol·L⁻¹) had the significant inhibition effects on cell proliferation of the TGF-β induced LX-2 cells(P<0.05). The cell apoptosis rate of curcumin derivative B group was higher than the model group(P<0.05). Collagen I, Fibronectin, Vimentin, α-SMA, TGFβR1 and TIMP-1 protein expression levels in curcumin group were lower, while the protein expression level of MMP-9 was higher(P<0.05). The protein expression levels of Collagen I, Collagen IV, Fibronectin, Vimentin, α-SMA, TIMP-1 and TIMP-2 in curcumin derivative A group were lower, while the protein expression level of MMP-2 was higher(P<0.05). The protein expression levels of Collagen I, Collagen IV, Fibronectin, Vimentin, α-SMA, PDGFRβ, TGFβR1, TGFβR2, TIMP-1 and TIMP-2 in curcumin derivative B group were lower, while the protein expression level of MMP-2 was higher(P<0.05). The gene transcription levels of Collagen I, Fibronectin, α-SMA and TIMP-1 in curcumin group were lower(P<0.05). The gene transcription levels of Collagen I, Fibronectin and α-SMA in curcumin derivative A and B groups were lower(P<0.05).
CONCLUSION Curcumin and its derivatives A and B inhibit the abnormal activation and proliferation of TGF-β-induced LX-2 cells, inhibit the excessive secretion and accumulation of its extracellular matrix components, and promote its degradation, thus playing an anti-fibrotic effect in vitro, especially the curcumin derivative B.

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