2,5-二氯-N-(2-甲基-4-硝基苯基)苯磺酰胺促进脂肪间充质干细胞向肝细胞分化

2,5-dichloro-N-(2-methyl-4-nitrophenyl)benzenesulfonamide promotes the differentiation of adipose mesenchymal stem cells into hepatocytes

目的:探讨2,5-二氯-N-(2-甲基-4-硝基苯基)苯磺酰胺(FH535)对脂肪间充质干细胞(ADSCs)向肝细胞分化的作用。方法:人ADSCs取自2021年3月新疆医科大学第一附属医院整形科吸脂患者皮下脂肪组织,分为肝细胞诱导分化组(HDM组)和FH535处理组(HDM+FH535组),糖原染色、免疫荧光及实时荧光定量聚合酶链反应(RT-qPCR)等检测分化效率。组间比较采用独立样本 t检验。 结果:FH535处理组诱导的肝细胞(ihep)百分比在诱导后第3天[(13.184±0.822)%比(8.680±0.273)%, t=11.630, P< 0.01]、第7天[(62.424±8.010)%比(20.736±4.790)%, t=9.988, P< 0.01]、第14天[(75.164±12.118)%比(25.132±2.235)%, t=9.079, P< 0.01]、第21天[(81.162±4.519)%比(51.622±8.362)%, t=6.950, P< 0.01]显著高于HDM组;FH535处理组ihep数量在诱导后第7天(98.600±7.436比62.200±14.567, t=4.977, P< 0.01)、第14天(149.600±16.682比56.200±4.207, t=12.140, P< 0.01)、第21天(201.600±27.501比137.000±25.000, t=3.887, P< 0.01)显著高于HDM组;FH535处理组糖原面积在诱导后第14天[(4 655.952±2 079.807) μm 2比(1 360.518±360.849) μm 2, t=3.491, P< 0.01]、第21天[(6 244.170±1 827.871) μm 2比(3 799.554±1 395.081) μm 2, t=2.377, P< 0.05]显著高于HDM组;FH535处理组肝脏基因表达水平在诱导分化的第14天[白蛋白(ALB)(44.525±7.795比19.251±9.497, t=3.563, P< 0.05)、转铁蛋白(Transferrin)(37.906±13.201比8.160±2.764, t=3.820, P< 0.05)、肝细胞核因子-1B(HNF-1B)(7.229±1.900比3.147±0.232, t=3.695, P< 0.05)、去唾液酸糖蛋白受体1(ASGPR1)(9.474±1.006比5.263±2.198, t=3.018, P< 0.05)、凝血酶-抗凝血酶复合物(TAT)(6.921±2.529比2.085±0.409, t=3.269, P< 0.05)、细胞角蛋白8(CK8)(7.707±3.049比0.807±0.143, t=3.916, P< 0.05)]显著高于HDM组。 结论:FH535可显著促进ADSCs的成肝细胞分化速率。

Objective:To investigate the effect of 2,5-dichloro-N-(2-methyl-4-nitrophenyl)benzenesulfonamide(FH535)on the differentiation of adipose-derived mesenchymal stem cells(ADSCs)into hepatocytes.Methods:Human ADSCs were obtained from subcutaneous adipose tissue of liposuction patients in the Department of Plastic Surgery of the First Affiliated Hospital of Xinjiang Medical University in March 2021.ADSCs were divided into hepatocyte differentiation group(HDM group)and FH535 group(HDM+FH535 group).Glycogen staining,Immunofluorescence and real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)were used to detect the differentiation efficiency.The independent-sample t test was used for comparison between groups.Results:The percentage of induced hepatocyte(ihep)in FH535 group at differentiation Day 3[(13.184±0.822)%vs.(8.680±0.273)%,t=11.630,P<0.01],Day 7[(62.424±8.010)%vs.(20.736±4.790)%,t=9.988,P<0.01],Day 14[(75.164±12.118)%vs.(25.132±2.235)%,t=9.079,P<0.01],Day 21[(81.162±4.519)%vs.(51.622±8.362)%,t=6.950,P<0.01]was significantly higher than that in HDM group.The number of ihep in FH535 group at differentiation Day 7(98.600±7.436 vs.62.200±14.567,t=4.977,P<0.01),Day 14(149.600±16.682 vs.56.200±4.207,t=12.140,P<0.01),Day 21(201.600±27.501 vs.137.000±25.000,t=3.887,P<0.01)was significantly higher than that in HDM group.Glycogen area in FH535 group at differentiation Day 14[(4655.952±2079.807)μm 2 vs.(1360.518±360.849)μm 2,t=3.491,P<0.01],Day 21[(6244.170±1827.871)μm 2 vs.(3799.554±1395.081)μm 2,t=2.377,P<0.05]was significantly higher than that in HDM group.The expression levels of hepatic gene in FH535 group at differentiation Day 14[albumin(ALB)(44.525±7.795 vs.19.251±9.497,t=3.563,P<0.05),Transferrin(37.906±13.201 vs.8.160±2.764,t=3.820,P<0.05),hepatocyte nuclear factor-1B(HNF-1B)(7.229±1.900 vs.3.147±0.232,t=3.695,P<0.05),asialoglycoprotein receptor 1(ASGPR1)(9.474±1.006 vs.5.263±2.198,t=3.018,P<0.05),thrombin-antithrombin complex(TAT)(6.921±2.529 vs.2.085±0.409,t=3.269,P<0.05),cytokeratin 8(CK8)(7.707±3.049 vs.0.807±0.143,t=3.916,P<0.05)]was significantly higher than that in HDM group.Conclusion:FH535 can significantly promote the differentiation of ADSCs into hepatocytes.