CAJ | 학술논문

目的：观察大鼠肝脏前体细胞系（WB-F344）对肝星状细胞系（HSC-T6）活化及细胞外基质的影响。方法将慢病毒-GFP 空白载体转染的 HSC-T6与 WB-F344按1∶1与1∶2比例直接共培养3 d，使用流式细胞仪对其进行分选，慢病毒-GFP 空白载体转染的 HSC-T6单独培养作为对照，观察 HSC-T6活化指标及细胞外基质相关指标表达的变化。结果经流式细胞仪检验慢病毒-GFP 空白载体转染 HSC-T6的效率可达近90％；与 WB-F344直接共培养3 d 后，HSC-T6的细胞形态与对照组相比无差别；同时利用 GFP 对各组 HSC-T6细胞分选后，经流式细胞仪检验纯度可达94％；对分选后的细胞进行分析发现，直接共培养组 HSC-T6细胞活化标记物α-平滑肌肌动蛋白（α-SMA）在基因水平（1∶1与1∶2组分别是对照组的0．66与0．61倍，P＜0．05）和蛋白水平（1∶1与1∶2组分别是对照组的0．61倍与0．25倍，P＜0．05）的表达均下降；HSC-T6细胞的细胞外基质标记物 I 型胶原（Col-I）、基质金属蛋白酶-13（MMP-13）、金属蛋白酶组织抑制剂-1（TIMP-1）的表达与对照组比较差异无统计学意义。结论WB-F344细胞可以抑制 HSC-T6细胞的活化，但在一定时间内对其细胞外基质的表达没有影响。
목적：관찰대서간장전체세포계（WB-F344）대간성상세포계（HSC-T6）활화급세포외기질적영향。방법장만병독-GFP 공백재체전염적 HSC-T6여 WB-F344안1∶1여1∶2비례직접공배양3 d，사용류식세포의대기진행분선，만병독-GFP 공백재체전염적 HSC-T6단독배양작위대조，관찰 HSC-T6활화지표급세포외기질상관지표표체적변화。결과경류식세포의검험만병독-GFP 공백재체전염 HSC-T6적효솔가체근90％；여 WB-F344직접공배양3 d 후，HSC-T6적세포형태여대조조상비무차별；동시이용 GFP 대각조 HSC-T6세포분선후，경류식세포의검험순도가체94％；대분선후적세포진행분석발현，직접공배양조 HSC-T6세포활화표기물α-평활기기동단백（α-SMA）재기인수평（1∶1여1∶2조분별시대조조적0．66여0．61배，P＜0．05）화단백수평（1∶1여1∶2조분별시대조조적0．61배여0．25배，P＜0．05）적표체균하강；HSC-T6세포적세포외기질표기물 I 형효원（Col-I）、기질금속단백매-13（MMP-13）、금속단백매조직억제제-1（TIMP-1）적표체여대조조비교차이무통계학의의。결론WB-F344세포가이억제 HSC-T6세포적활화，단재일정시간내대기세포외기질적표체몰유영향。
Objective To explore the effects of HPCs (WB-F344)on activation and extracellular matrix (ECM)of HSCs (HSC-T6).Methods Lentivirus-GFP transfected HSC-T6 was directly cocultured with WB-F344 at 1 ∶1 and 1 ∶2 ratios for 3 days.Mixed cells were sorted by flow cytometry based on the presence or absence of GFP.Transfected HSC-T6 cultured alone was used as control.Expression of activation marker α-smooth muscle actin (α-SMA)and ECM markers collage I (Col-I),matrix metal proteinase-13 (MMP-13),tissue inhibitor of metalloproteinase-1 (TIMP)-1 in HSC-T6 were observed by Western blot and real-time PCR.Results After 3 days of coculture with WB-F344,morphological changes of HSC-T6 remained unchanged in direct coculture groups.Transfection efficiency of Lentivirus-GFP was nearly 90% and the purity of sorted HSC-T6 was 94%.WB-F344 directly cocultured with HSC-T6 had reduced α-SMA expression of HSC-T6 in mRNA and protein level.However,there was no difference in ECM expression between control group and direct cocul-ture groups.Conclusion WB-F344 can inhibit the activation of HSC-T6,but it had no influence on expression of ECM.