中华临床感染病杂志
中華臨床感染病雜誌
중화림상감염병잡지
CHINESE JOURNAL OF CLINICAL INFECTIOUS DISEASES
2008年
1期
15-18
,共4页
哈明昊%饶慧瑛%刘峰%费然%从旭%陈红松%魏来
哈明昊%饒慧瑛%劉峰%費然%從旭%陳紅鬆%魏來
합명호%요혜영%류봉%비연%종욱%진홍송%위래
肝炎病毒,乙型%肝纤维化%肝星状细胞%基质金属蛋白酶-2%金属蛋白酶组织抑制剂-1%共同培养
肝炎病毒,乙型%肝纖維化%肝星狀細胞%基質金屬蛋白酶-2%金屬蛋白酶組織抑製劑-1%共同培養
간염병독,을형%간섬유화%간성상세포%기질금속단백매-2%금속단백매조직억제제-1%공동배양
Hepatitis B virus%Liver fibrosis%Hepatic stellate cell%Matrix metalloproteinase-2%Tissue inhibitor of metalloproteinase-1%Co-culture
目的 研究HepG2.2.15细胞株在体外能否促进肝星状细胞(HSC)中肝纤维化相关因子的表达,进而探索HBV促肝细胞纤维化的机制.方法 将HepG2和HepG2.2.15细胞株分别在体外与HSC共培养,以单独培养的HSC为对照组.取培养后24、48和72 h 3个时间点,以实时定量PCR检测HSC中基质金属蛋白酶-2(MMP-2)和金属蛋白酶组织抑制剂-1(TIMP-1)mRNA的表达;以免疫印迹法定量检测HSC中MMP-2和TIMP-1的表达.结果 与对照组和与HepG2共培养的HSC比较,与HepG2.2.15细胞株共培养的HSC中MMP-2和TIMP-1 mRNA的表达明显增高,以72 h的差异最为显著(F值分别为11.91和23.13,P值分别为0.008和0.001);与HepG2.2.15共培养的HSC中MMP-2和TIMP-1蛋白的表达亦明显增高(F值分别为20.70和6.54,P值分别为0.002和0.03).结论 与HepG2.2.15细胞株共培养后,HSC中肝纤维化相关因子的表达明显增强.体外试验证明HBV具有诱导肝细胞纤维化的重要作用.
目的 研究HepG2.2.15細胞株在體外能否促進肝星狀細胞(HSC)中肝纖維化相關因子的錶達,進而探索HBV促肝細胞纖維化的機製.方法 將HepG2和HepG2.2.15細胞株分彆在體外與HSC共培養,以單獨培養的HSC為對照組.取培養後24、48和72 h 3箇時間點,以實時定量PCR檢測HSC中基質金屬蛋白酶-2(MMP-2)和金屬蛋白酶組織抑製劑-1(TIMP-1)mRNA的錶達;以免疫印跡法定量檢測HSC中MMP-2和TIMP-1的錶達.結果 與對照組和與HepG2共培養的HSC比較,與HepG2.2.15細胞株共培養的HSC中MMP-2和TIMP-1 mRNA的錶達明顯增高,以72 h的差異最為顯著(F值分彆為11.91和23.13,P值分彆為0.008和0.001);與HepG2.2.15共培養的HSC中MMP-2和TIMP-1蛋白的錶達亦明顯增高(F值分彆為20.70和6.54,P值分彆為0.002和0.03).結論 與HepG2.2.15細胞株共培養後,HSC中肝纖維化相關因子的錶達明顯增彊.體外試驗證明HBV具有誘導肝細胞纖維化的重要作用.
목적 연구HepG2.2.15세포주재체외능부촉진간성상세포(HSC)중간섬유화상관인자적표체,진이탐색HBV촉간세포섬유화적궤제.방법 장HepG2화HepG2.2.15세포주분별재체외여HSC공배양,이단독배양적HSC위대조조.취배양후24、48화72 h 3개시간점,이실시정량PCR검측HSC중기질금속단백매-2(MMP-2)화금속단백매조직억제제-1(TIMP-1)mRNA적표체;이면역인적법정량검측HSC중MMP-2화TIMP-1적표체.결과 여대조조화여HepG2공배양적HSC비교,여HepG2.2.15세포주공배양적HSC중MMP-2화TIMP-1 mRNA적표체명현증고,이72 h적차이최위현저(F치분별위11.91화23.13,P치분별위0.008화0.001);여HepG2.2.15공배양적HSC중MMP-2화TIMP-1단백적표체역명현증고(F치분별위20.70화6.54,P치분별위0.002화0.03).결론 여HepG2.2.15세포주공배양후,HSC중간섬유화상관인자적표체명현증강.체외시험증명HBV구유유도간세포섬유화적중요작용.
Objective To investigate the effect of HBV on the expression of fibrosis-related factors in hepatic stellate cells(HSC)and its relation with liver fibrosis.Methods HSCs were co-cultured with HepG2 or HepG2.2.15 in vitro and HSCs cultured alone served as the control.The mRNA expression of matrix metalloproteinase(MMP)-2 and tissue inhibitor of metalloproteinase(TIMP)-1 was detected by realtime PCR.The protein expression of MMP-2 and TIMp-1 was detected by Western-blot.Results Compared with the control and the HSCs co-cultured with HepG2,the expression of MMP-2 and TIMP-1 mRNA in HSCs co-cultured with HepG2.2.15 was increased remarkably and the most significant difference was found at 72 h(F=11.91,23.13;P=0.008,0.001);the expression of MMP-2 and TIMP-1 protein in HSCs co-cuhured with HepG2.2.15 was also increased remarkably and the most significant difference was found at 72 h(F=20.70,6.54;P=0.002,0.003)too.Conclusion The expression of fibrosis-related factors in HSCs increased significantly after co-cultured with HepG2.2.15,which suggests that HBV could promote liver fibrosis.
