摘要
目的:通过观察肿瘤坏死因子α(TNF-α)和转化生长因子β1(TGF-β1)两种细胞因子所致体外培养的肝细胞损伤时胰岛素样生长因子结合蛋白2、4(IGFBP2、IGFBP4)的表达变化,探讨IGFBP2和IGFBP4在肝损伤中的作用机制。方法:对人肝细胞株HL-7702分别给予不同浓度的TNF-α、TGF-β1两种处理因素作用24 h,采用免疫细胞化学染色法观察其IGFBP2和IGFBP4的表达变化,再在相同处理培养条件下用MTT比色法检测两种细胞因子对肝细胞抑制率的影响。根据MTT及预实验结果选择TNF-α20μg/L作用于人肝细胞株48h,采用Annexin-Ⅴ/PI双染色流式细胞分析法和TUNEL法检测肝细胞凋亡发生率。结果:与对照组相比,各处理组IGFBP2、IGFBP4的表达均显著增加(P<0.05),其中在TNF-α20μg/L组和TGF-β14μg/L组表达最强,且与肝细胞抑制率呈正相关关系(P<0.05 orP<0.01)。TNF-α20μg/L处理48 h后肝细胞凋亡率明显增加(P<0.01)。结论:IGFBP2、IGFBP4参与了肝细胞的损伤过程,在TNF-α、TGF-β1介导的肝细胞损伤中具有重要作用。
AIM: To explore the mechanism of IGFBP2 and IGFBP4 in hepatocytes with injury induced by TNF - α and TGF - β1. METHODS: Human hepatocyte line ( HL - 7702) was cultured in vitro and treated with TNF - α or TGF -β1 for 24 h. The expression of IGFBP2 and IGFBP4 was detected by immunochemistry staining. The inhibition ratio of hepatocytes was detected by MTr assay. HL - 7702 was treated with TNF -α at concentration of 20 μg/L for 48 h, then the apoptosis of hepatocytes was detected by both Annexin - V/PI and TUNEL assay. RESULTS : The expression of IG- FBP2 and IGFBP4 in TNF -α or TGF - β1 treated groups was significantly higher than that in control group (P 〈 0. 05 ). The positive staining of IGFBP2 and IGFBP4 in TNF - α (20 μg/L) treated groups or TGF - β1 (4 μg/L) treated groups was the strongest among all groups. A positive correlation was found between IGFBP2 or IGFBP4 and inhibitory ratio of hepatocytes ( P 〈 0.05 or P 〈 0. 01 ). Compared with normal control group, the percentage of apeptosis markedly increased in TNF-α (20 μg/L) treated group (P 〈 0. 01 ). CONCLUSION: IGFBP2 and IGFBP4 involved in the injury process in hepatocytes, indicating an important role in injury of hepatocytes induced by TNF - α or TGF - β1.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2009年第1期137-143,共7页
Chinese Journal of Pathophysiology
基金
教育部"新世纪优秀人才支持计划"资助项目(No.2006348)
山西省自然科学基金资助项目(No.20051092)
山西省教育厅青年学术带头人基金资助项目(No.2005017)