摘要
目的 采用肝星状细胞（HSCs）株LX-2探讨不同浓度奥曲肽对该细胞生物学活性的影响及其在肝纤维化形成中的作用机制.方法 应用噻唑蓝（MTT）法检测奥曲肽对LX-2细胞增殖的影响;反转录-聚合酶链反应（RT-PCR）法分析用奥曲肽处理后,细胞中转化生长因子-β1（TGF-β1）、α-平滑肌肌动蛋白（α-SMA）、Smad-4及c-Jun的转录水平;酶联免疫吸附试验（ELISA）法检测LX-2培养上清中Ⅰ型胶原的水平;小干扰RNA （siRNA）和染色质免疫共沉淀技术检测c-Jun对TGF-β1表达的调控及其对TGF-β启动子的结合.结果 奥曲肽能抑制LX-2细胞的增殖能力,细胞增殖以浓度依赖的方式显著降低（P＜0.05）,当奥曲肽浓度为10-7 nmol/L和10-5 nmol/L时,其抑制率分别为27.5％和49.8％,奥曲肽组与对照组比较,能显著降低TGF-β1、α-SMA、Smad-4及c-Jun的表达（P＜0.05）,同时上清中Ⅰ型胶原水平明显减少（P＜0.05）,沉默c-Jun的表达后,TGF-β1的表达显著降低.结论 奥曲肽可通过抑制HSCs的增殖,降低α-SMA、Smad-4及c-Jun的表达,进而调控TGF-β1,减少Ⅰ型胶原的分泌,从而减缓纤维化的发生,对降低门静脉高压可能有重要的临床意义.
Objective To investigate the influence of octreotide to the activity of LX-2,a hepatic stellate cell line,at different concentration,and explore its effect and mechanism in the process of liver fibrosis.Methods Methyl thiazol tetrazolium （MTT） assay was conducted for the influence of octreotide to the proliferation of LX-2.The mRNA levels of transforming growth factor-β1 （TGF-β1）,α-smooth muscle actin （α-SMA）,smad-4 and c-Jun under octreotide were detected by reverse transcriptase-polymerase chain reaction （RT-PCR）.The content of type-Ⅰ collagen in the supernant of cultured LX-2 was assayed by enzyme linked immunosorbent assay （ELISA）.Whether c-Jun regulates TGF-β through binding to its promoter was monitored based on small interfering RNA （siRNA） and chromatin immunoprecipitation （ChIP） technologies.Results The proliferation of LX-2 was inhibited obviously by octreotide in a concentration-dependent manner,the inhibition rates of which were 27.5% and 49.8% at 10-7 nmol/L and 10-5 nmol/L.Compared with control,the transcriptions of TGF-β1,α-SMA,smad-4 and c-Jun were down-regulated significantly under octreotide （P 〈 0.05）,and the content of type-Ⅰ collagen reduced significantly （P 〈 0.05）.The expression of TGF-β1 reduced significantly under interference of c-Jun.Conclusion Octreotide could inhibit the proliferation of LX-2,reduce the expressions of TGF-β1,α-SMA,Smad-4 and c-Jun,regulate TGF-β1,and reduce the secretion of type-Ⅰ collagen,and retard the occurrence of fibrosis,provided important clinical significance to control portal hypertension.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2015年第3期508-510,共3页
Chinese Journal of Experimental Surgery
基金
山东省科技发展计划资助项目（2010G0020223）
关键词
奥曲肽
肝星状细胞
肝纤维化
Octreotide
Hepatic stellate cell
Liver fibrosis
