脯氨酰寡肽酶抑制剂体外对肝星状细胞凋亡、增殖和肝纤维化相关基因表达的影响*

doi:10.3969/j.issn.1672-5069.2018.03.018

摘要/Abstract

摘要： 目的体外探讨肝星状细胞（HSC）脯氨酰寡肽酶（POP）的生物学功能,以进一步阐述其在肝脏炎症和纤维化发生发展中的作用。方法取HSC-T6细胞,经不同浓度的POP抑制剂（S17092）处理,采用ELISA法检测HSC-T6细胞N-乙酰基-丝氨酸-天冬氨酸-赖氨酸-脯氨酸（Ac-SDKP）水平,使用流式细胞术和CCK8检测HSC-T6凋亡和增殖水平,采用实时定量-PCR法检测相关炎症和肝纤维化基因转化生长因子-β1（TGF-β1）、α-平滑肌肌动蛋白（α-SMA）、单核细胞趋化蛋白-1（MCP-1）、Ⅰ型胶原（Col I）水平,采用Western blot法检测相关蛋白（POP、TGF-β1、α-SMA、Smad7、p-Smad2/3、PPAR-γ）表达。结果与对照组比,POP抑制剂干预组细胞内Ac-SDKP水平显著下降（P<0.05）;POP被抑制后对HSC-T6凋亡没有显著的影响（P>0.05）,但可抑制其增殖（P<0.05）;与对照组比,抑制剂组HSC内α-SMA蛋白表达显著升高（P<0.05）,而Smad7和PPAR-γ蛋白表达显著下降（P均<0.05）;抑制剂组HSC内MCP-1和α-SMA mRNA水平显著上调（P均<0.05）。结论 POP在肝星状细胞内可能发挥重要的抗炎抗纤维化作用,其机制可能与其调节Ac-SDKP及Smad7和PPAR-γ的表达有关。

关键词: HSC-T6细胞, 脯氨酰寡肽酶, 转化生长因子-β-Smad蛋白信号通路, 过氧化物酶体增殖剂激活受体-&#x003b3, 体外

Abstract: Objective To investigate the inhibition of proliferation and regulation of heaptic fibrosis-related genes in HSC-T6 cells by prolyloligopeptidase（POP） inhibitor in vitro. Methods The HSC-T6 cells were cultured,and the POP activity was inhibited by increasing concentrations of POP inhibitor（S17092）. The intracellular level of N-acetyl- seryl-aspartyl-lysyl-proline（Ac-SDKP） was determined by ELISA. The cell apoptosis and cell proliferation were detected by flow cytometry and CCK-8 assay,respectively. The mRNA levels of TGF-β1,α-SMA,monocyte chemoattractant protein 1（MCP-1） and collagen I（Col I） were detected by realtime-PCR,and the POP,TGF-β1,α-SMA, Smad7,p-Smad2/3 and PPAR-γ expression were detected by Western blot. Results The intracellular Ac-SDKP level decreased significantly after co-culture with S17092,the POP inhibitor（P<0.05） as compared to that in the control; the proliferation of HSC-T6 cells was significantly inhibited（P<0.05） and the cell apoptosis was not influenced obviously after S17092 intervention（P>0.05）;the Smad7（P<0.05） and PPAR-γ（P<0.05） protein levels were significantly down-regulated,and the α-SMA protein level was significantly up-regulated（P<0.05） after S17092 intervention;the MCP-1（P<0.05） and α-SMA（P<0.05） mRNA level were significantly up-regulated. Conclusion POP plays a pivotal role in anti-inflammation and anti-fibrosis in HSC cells, which might be related to the regulation of Smad7 and PPAR-γ expression.

Key words: HSC-T6 cells, Prolyloligopeptidase, Transforming growth factor-&#x003b2, -Smad signaling, Peroxisome proliferator activated receptor-&#x003b3, In vitro

周达, 王晶, 李冰航, 孙超, 陈源文, 范建高. 脯氨酰寡肽酶抑制剂体外对肝星状细胞凋亡、增殖和肝纤维化相关基因表达的影响*[J]. 实用肝脏病杂志, 2018, 21(3): 388-392.

Zhou Da, Wang Jing, Li Binghang, et al. Inhibition of proliferation and regulation of heaptic fibrosis-related genes in HSC-T6 cells by prolyloligopeptidase inhibitor in vitro[J]. JOURNAL OF PRACTICAL HEPATOLOGY, 2018, 21(3): 388-392.

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