摘要
目的研究血红素加氧酶-1(HO-1)对肝窦内皮细胞(LSECs)增殖、迁移及促肝细胞增殖作用的影响。方法雄性6~8周龄无特定病原体级C57BL/6小鼠18只,行部分肝切除,组织免疫荧光检测术后0、2、4 d残肝内细胞增殖及HO-1表达。以携带HO-1基因的腺病毒转染LSECs细胞系(HO-1组),同时以空载体腺病毒转染和未转染细胞为对照。另外建立不同转染组LSECs与肝细胞系非接触共培养模型,评价HO-1表达对肝细胞的促增殖作用。Western印迹及实时定量聚合酶链反应检测各组细胞HO-1、分化抑制因子1(Id1)、肝细胞生长因子(HGF)、Wnt2蛋白和mRNA表达水平变化,EdU法检测各组细胞增殖,Transwell迁移实验检测细胞迁移能力变化。结果相比小鼠肝切除术后0 d,术后4 d再生肝内LSECs开始大量增殖并伴随LSECs内HO-1表达明显升高。HO-1组LSECs的EdU阳性率(27.20±4.80)%高于空载体组(12.47±3.30)%和未转染对照组(15.97±2.50)%,HO-1组LSECs迁移数量(258.70±36.56)个高于空载体组(122.00±38.16)个和对照组(107.70±30.01)个(均为5个200×视野),HO-1组HO-1、Id1、HGF、Wnt2蛋白及mRNA表达水平均高于空载体组与对照组,差异均有统计学意义(均P<0.05)。HO-1组LSECs共培养的肝细胞EdU阳性率(18.33±2.52)%高于空载体组(11.33±1.53)%与对照组(11.70±2.08)%,差异均有统计学意义(均P<0.05)。结论上调HO-1表达能够促进小鼠LSECs增殖、迁移以及增强LSECs对肝细胞的促增殖作用。
Objective To investigate the effects of heme oxygenase-1(HO-1)on hepatic sinusoidal endothelial cells(LSECs)proliferation,migration,and hepatocyte proliferation.Methods Eighteen male C57BL/6 mouse aged 6-8 weeks old were underwent partial hepatectomy.Cell proliferation and HO-1 expression in residual liver tissue were detected by immunofluorescence histochemistry at 0 d,2 d and 4 d after operation.In vitro,LSECs were transfected with adenovirus carrying HO-1 gene(HO-1 group),and the cells were transfected with empty vector adenovirus and the non-transfected cells were used as control.In addition,LSECs from different transfection groups were co-cultured with hepatocyte without contact to evaluate the effect of HO-1 expression on promoting hepatocyte proliferation.Western Blotting and RT-PCR were used to detect the protein and mRNA expression of HO-1,inhibitor of DNA binding and or differentiation(Id1),hepatocyte growth factor(HGF)and Wnt2.Cell proliferation was detected by EdU.The ability of cell migration was detected by Transwell migration assay.Results Compared with 0 d after hepatectomy,LSECs proliferation and HO-1 expression within LSECs were increased significantly at 4 d after surgery.EdU positive rate of LSECs in HO-1 group(27.20±4.80)%was higher than that in empty vector group(12.47±3.30)%and non-transfected group(15.97±2.50)%.The number of LSECs migration in HO-1 group(258.70±36.56)was higher than that in empty vector group(122.00±38.16)and non-transfected group(107.70±30.01).The protein and mRNA expression level of HO-1,Id1,HGF and Wnt2 in HO-1 group were higher than that in empty vector group and non-transfected group.EdU positive rate of hepatocytes that co-cultured with LSECs in HO-1 group(18.33±2.52)%was higher than that in empty vector group(11.33±1.53)%and non-transfected group(11.7±2.08)%.The differences were statistically significant(all P<0.05).Conclusion Up-regulation of HO-1 promoted LSECs proliferation and migration of,as well as up-regulation of HO-1 in LSECs enhanced the capacity
作者
袁博
明廷凤
瞿思铭
黄汉飞
曾仲
林杰
Yuan Bo;Ming Tingfeng;Qu Siming;Huang Hanfei;Zeng Zhong;Lin Jie(Organ Transplantation Center,First Affiliated Hospital of Kunming Medical University,Kunming 650032,China)
出处
《中华肝胆外科杂志》
CAS
CSCD
北大核心
2022年第7期536-541,共6页
Chinese Journal of Hepatobiliary Surgery
基金
云南省教育厅科学研究基金(2020Y0121)
国家自然科学基金(81960124)。
关键词
肝再生
内皮细胞
肝细胞
血红素加氧酶-1
Liver regeneration
Endothelial cells
Hepatocytes
Heme oxygenase-1
