补肾活血解毒中药对大鼠肝细胞及卵圆细胞增殖与功能的影响

Effect of chinese drugs for invigorating Kidney and Promoting Blood Circulation and Detoxification on Proliferation and Function of Rat Hepatocytes and Oval Cells

目的探讨补肾活血解毒中药含药血清对原代大鼠肝细胞及肝卵圆细胞增殖与功能的影响。方法采用2-乙酰氨基芴加肝脏三分之二切除术建立肝卵圆细胞增殖模型,通过原位二步胶原酶灌流法分离到单个肝脏细胞,再经过Percoll密度梯度离心分离到肝卵圆细胞。原代大鼠肝细胞/肝卵圆细胞各分为3组:普通血清组、肝衰竭血清组及含药血清组,分别于培养后第1、3、6、8d采用MTT法检测各组细胞增殖能力;干预培养72h后再常规培养24 h,然后采用ELISA法分别检测3组细胞上清液中白蛋白水平。结果含药血清组的原代肝细胞和肝卵圆细胞在培养后第1、3、6、8d的OD值均比相应时间点的普通血清组和肝衰竭血清组的OD值大(P<0.05);含药血清组的肝卵圆细胞培养上清液中白蛋白水平较普通血清组和肝衰竭血清组均高(P<0.05),含药血清组和肝衰竭血清组的原代肝细胞培养上清液中白蛋白水平均较普通血清组高(P<0.05),含药血清组和肝衰竭血清组的原代肝细胞培养上清液中白蛋白水平差异无统计学意义(P>0.05)。结论补肾活血解毒中药能促进原代大鼠肝细胞及肝卵圆细胞在体外增殖和白蛋白分泌。

Objective To investigate the effect of drug-containing serum with Traditional Chinese Medicine for tonifying kidney and activating blood circulation and detoxification on the proliferation and function of primary rat hepatocytes and hepatic oval cells.Methods Rat model for hepatic oval cell proliferation was establish with 2-acetylaminofluorenne and two third partial hepatectomy(2-AAF/PH).Discrete liver cells were isolated by two steps collagenase perfusion via portal vein,and the hepatic oval cells were separated by density gradient centrifugation of Percoll.The primary rat hepatocyte/hepatic oval cells were divides into three groups normal serum group,hepatic failure serum group and drug-containing serum group.The cells proliferation rate of every group were measured by MTT on 1 d,3 d,6 d,8 d,intervention culture for 72 hours and routine culture for 24 hours,the concentration of albumin in the culture medium were measured by ELISA.Results OD values of primary hepatocyte and hepatic oval cells in the drug-containing serum group were higher than those in the normal serum group and the hepatic failure serum group on1d,3d,6d,8d(P<0.05).The level of albumin in the liver oval cell culture in the drug-containing serum group was higher than that in the normal serum group and the hepatic failure serum group(P<0.05),and the albumin level in the primary liver cell culture of the drug-containing serum group and the hepatic failure serum group was all higher than that of the normal serum group(P<0.05).There was no significant difference in albumin levels in the primary hepatocyte culture between the drug-containing serum group and the hepatic failure serum group(P>0.05).Conclusion Serum with tonifying kidney and activating blood circulation and detoxification Chinese medicine can promote proliferation of primary rat hepatocyte and hepatic oval cells in vitro,and increase secretion of albumin.