中华肝胆外科杂志
中華肝膽外科雜誌
중화간담외과잡지
CHINESE JOURNAL OF HEPATOBILIARY SURGERY
2013年
9期
701-705
,共5页
俞富祥%宋才鑫%吴志伟%朱千东%张启瑜
俞富祥%宋纔鑫%吳誌偉%硃韆東%張啟瑜
유부상%송재흠%오지위%주천동%장계유
脂肪特异性蛋白27%基因%肝星状细胞%增殖%活化
脂肪特異性蛋白27%基因%肝星狀細胞%增殖%活化
지방특이성단백27%기인%간성상세포%증식%활화
Fat-specific protein 27%Genes%Hepatic stellate cells%Proliferation%Activation
目的 探讨脂肪特异性蛋白27(Fsp27)基因对活化态肝星状细胞(HSCs)增殖、活化的影响及其对纤维化相关基因的调节作用.方法 提取HSCs并培养.用实时定量PCR技术检测原代HSCs及活化HSCs中的Fsp27基因表达.构建携带Fsp27目的基因的慢病毒,转染活化的HSCs并继续培养72 h.通过CCK-8比色法检测Fsp27基因对HSCs增殖的影响.Western blot检测HSCsα-肌动蛋白(α-SMA)的表达,了解HSCs的活化状态.实时定量PCR技术研究Fsp27基因对HSCs纤维化相关基因表达的影响.结果 成功分离原代HSCs,Fsp27基因在原代HSCs及活化HSCs中表达差异显著(P＜0.01).活化HSCs成功转染携带Fsp27目的基因的慢病毒后继续培养72 h,与对照组比较,HSCs的活化与增殖受到明显抑制(P＜0.05)；Fsp27基因促进MMP-2的表达(P＜0.05),降低了TIMP-1及TGF-β1的表达(P＜0.05).结论 Fsp27基因具有抑制HSCs活化增殖的潜能,Fsp27基因可调节纤维化相关基因的表达.Fsp27基因的作用可能与维持HSCs静息状态细胞表型有关.
目的 探討脂肪特異性蛋白27(Fsp27)基因對活化態肝星狀細胞(HSCs)增殖、活化的影響及其對纖維化相關基因的調節作用.方法 提取HSCs併培養.用實時定量PCR技術檢測原代HSCs及活化HSCs中的Fsp27基因錶達.構建攜帶Fsp27目的基因的慢病毒,轉染活化的HSCs併繼續培養72 h.通過CCK-8比色法檢測Fsp27基因對HSCs增殖的影響.Western blot檢測HSCsα-肌動蛋白(α-SMA)的錶達,瞭解HSCs的活化狀態.實時定量PCR技術研究Fsp27基因對HSCs纖維化相關基因錶達的影響.結果 成功分離原代HSCs,Fsp27基因在原代HSCs及活化HSCs中錶達差異顯著(P＜0.01).活化HSCs成功轉染攜帶Fsp27目的基因的慢病毒後繼續培養72 h,與對照組比較,HSCs的活化與增殖受到明顯抑製(P＜0.05)；Fsp27基因促進MMP-2的錶達(P＜0.05),降低瞭TIMP-1及TGF-β1的錶達(P＜0.05).結論 Fsp27基因具有抑製HSCs活化增殖的潛能,Fsp27基因可調節纖維化相關基因的錶達.Fsp27基因的作用可能與維持HSCs靜息狀態細胞錶型有關.
목적 탐토지방특이성단백27(Fsp27)기인대활화태간성상세포(HSCs)증식、활화적영향급기대섬유화상관기인적조절작용.방법 제취HSCs병배양.용실시정량PCR기술검측원대HSCs급활화HSCs중적Fsp27기인표체.구건휴대Fsp27목적기인적만병독,전염활화적HSCs병계속배양72 h.통과CCK-8비색법검측Fsp27기인대HSCs증식적영향.Western blot검측HSCsα-기동단백(α-SMA)적표체,료해HSCs적활화상태.실시정량PCR기술연구Fsp27기인대HSCs섬유화상관기인표체적영향.결과 성공분리원대HSCs,Fsp27기인재원대HSCs급활화HSCs중표체차이현저(P＜0.01).활화HSCs성공전염휴대Fsp27목적기인적만병독후계속배양72 h,여대조조비교,HSCs적활화여증식수도명현억제(P＜0.05)；Fsp27기인촉진MMP-2적표체(P＜0.05),강저료TIMP-1급TGF-β1적표체(P＜0.05).결론 Fsp27기인구유억제HSCs활화증식적잠능,Fsp27기인가조절섬유화상관기인적표체.Fsp27기인적작용가능여유지HSCs정식상태세포표형유관.
Objective To investigate the Fsp27 gene's influence on the regulation of hepatic stellate cells (HSCs) in vitro.Methods HSCs were isolated from rat liver,the Fsp27 gene was detected in primary HSCs,and activated HSCs were detected by RT-qPCR.After 72 h of Fsp27 transduction through a lentivirus expressing Fsp27 (pLV-Fsp27),the proliferation of HSCs was tested by the CCK-8 test kit,smooth muscle α-actin (α-SMA) expression of HSCs was tested by Western blot,and the fibrosis-related genes were tested by RT-qPCR.Results The HSCs were isolated and cultured successfully,and the Fsp27 genetic difference between primary and activated HSCs was significant (P＜0.01).After coculture for 72 h,Fsp27 inhibited the proliferation and activation of HSCs (P＜0.05).Fsp27 can enhance expression of the MMP-2 gene and down-regulate expression of the TIMP-1 and TGF-β1 gene in activated HSCs (P＜0.05).Conclusion The Fsp27 gene can inhibit the proliferation and activation of HSCs,regulate the expression of fibrosis-related genes,and may play an important role in maintaining the quiescent phenotype of HSCs.
