摘要
目的探讨肝衰竭患者血浆对永生化人肝细胞(immortalized human hepatocytes,IHH)增殖和肝细胞功能蛋白谷氨酰胺合成酶(glutamine synthetase,GS)、谷胱甘肽-S-转移酶(glutathione-S-transferase,GST)、尿苷二磷酸葡萄糖醛酸基转移酶(uridine diphosphate glucuronosyltransferase,UGT)和白蛋白(albumin,Alb)表达的影响。方法 IHH细胞分为观察组和对照组,对照组细胞采用含体积分数5%胎牛血清的培养基进行培养,观察组细胞采用含体积分数5%肝衰竭患者血浆的培养基进行培养,应用倒置显微镜观察2组细胞生长状态,采用CCK8法测定2组细胞第1、3、5、7、9天增殖情况,并绘制生长曲线;采用Western blot法检测2组细胞中肝细胞功能蛋白GS、GST、UGT、Alb及肝细胞生长因子(hepatocyte growth factor,HGF)蛋白表达水平;采用实时荧光定量PCR检测2组细胞中HGF mRNA表达水平。结果2组细胞形态正常,呈梭形贴壁生长,观察组细胞第5、7、9天时细胞增殖吸光度值(1.44±0.08、1.87±0.11、1.95±0.06)明显高于对照组(1.26±0.06、1.62±0.08、1.78±0.07)(P<0.05);观察组细胞Alb表达水平(1.32±0.07)高于对照组(0.94±0.05)(P<0.05),GS、GST、UGT表达水平与对照组比较差异无统计学意义(P>0.05);观察组细胞HGF mRNA和蛋白(7.940±0.203、1.17±0.05)表达水平均高于对照组(1.020±0.135、0.97±0.04)(P<0.05)。结论肝衰竭患者血浆可促进IHH细胞增殖,上调肝细胞功能蛋白Alb表达,同时对GS、GST、UGT蛋白表达无明显影响;其促增殖机制可能与HGF表达上调有关。
Objective To investigate the influence of the plasma from liver failure patients on the proliferation of immortalized human hepatocytes(IHH)and the expressions of hepatocyte function proteins as glutamine synthetase(GS),glutathione-S-transferase(GST),uridine diphosphate glucuronosyltransferase(UGT)and albumin(Alb)in patients with liver failure.Methods IHH cells were divided into observation group and control group.Control group was cultured with the medium containing 5% fetal bovine serum,and observation group was cultured with the medium containing 5% plasma from liver failure patients.The cell growth was observed by inverted microscope.The proliferation was determined by CCK8 assay on the 1 st,3 rd,5 th,7 th and 9 th days to draw growth curve.The expressions of GS,GST,UGT,Alb and hepatocyte growth factor(HGF)protein were detected by Western blot method.The expression of HGF mRNA was tested by real-time fluorescence quantitative PCR in both groups.Results The morphology was normal and in spindle-shaped adherent growth in two groups.The absorbance values of cell proliferation were significantly higher in observation group(1.44±0.08,1.87±0.11,1.95±0.06)than those in control group(1.26±0.06,1.62±0.08,1.78±0.07)on the 5 th,7 th and 9 th days(P<0.05).The Alb level was significantly higher in observation group(1.32±0.07)than that in control group(0.94±0.05)(P<0.05).There were no significant differences in the expressions of GS,GST and UGT proteins between two groups(P>0.05).The mRNA and protein of HGF were significantly higher in observation group(7.940±0.203,1.17±0.05)than those in control group(1.020±0.135,0.97±0.04)(P<0.05).Conclusion The plasma from liver failure patients can promote the proliferation of IHH and up-regulate the expression of Alb,while it shows no significant effect on the expressions of GS,GST and UGT proteins.The proliferation mechanism may be correlated with the up-regulation of HGF expression.
出处
《中华实用诊断与治疗杂志》
2017年第10期971-974,共4页
Journal of Chinese Practical Diagnosis and Therapy
基金
国家高技术发展研究计划(863计划)(2006AA02Z482)
关键词
肝衰竭
血浆
永生化人肝细胞
生物人工肝
肝细胞生长因子
Liver failure
plasma
immortalized human hepatocytes
bioartificial liver
hepatocyte growth factor
