Received 1999205207 Revised 1999207219

    3

    Supported by the National Natural Science Foundation of China (No139470725)

    3 3

    Corresponding author. Tel : (0411) 4720032 , E 2mail : zouy@mail1dlmedu1edu1cn

    生理学报 , Feb. 2000 , 52 (1) : 59～63 59

    Acta Physiologica Sinica

    肝细胞生长因子对四氯化碳损伤

    肝细胞的保护作用及相关基因表达3

    邹 原3 3

    , 宫德正, 梅懋华, 张万琴

    (大连医科大学生理教研室, 大连 116027)

    摘 要: 利用无血清原代培养大鼠肝细胞 , 观察重组人肝细胞生长因子( rhHGF)对 CCl4 染毒肝细胞的保护作

    用。结果表明: (1) rhHGF (5 ng/ ml)预处理后可显著提高 CCl4 (15 mmol/ L)染毒肝细胞存活率 , 降低细胞内丙氨酸

    氨基转移酶(ALT) 、K+

    的漏出; (2) 表皮生长因子(EGF , 50 ng/ ml)和 rhHGF (5 ng/ ml)合用预处理肝细胞 , CCl4 染

    毒后细胞内ALT、K+

    漏出较 rhHGF和 EGF单独保护组进一步降低; (3)大鼠肝部分切除和 CCl4 (50 % , 215 ml/ kg

    bw)染毒后 , 再生肝内 HGF基因及其受体基因/ c2met 的表达分别较假手术和盐水对照组显著升高。结果提示 ,rhHGF对 CCl4 染毒大鼠肝细胞具有保护作用; EGF和 rhHGF有协同保护作用; HGF及其受体的表达在肝脏再生及

    修复中可能有重要作用。

    关键词: 重组人肝细胞生长因子; 表皮生长因子; c2met 基因; 四氯化碳; 肝细胞原代培养

    学科分类号: Q485 ; R33314

    Protective effect of hepatocyte growth factor on hepatocyte poisoning by

    carbon tetrachloride and related gene expression in rats

    3

    ZOU Yuan , GONGDe2Zheng , MEI Mao2Hua , ZHANGWan2Qin

    ( Department of Physiology , Dalian Medical University , Dalian 116027)

    Abstract : Hepatocyte growth factor (HGF) , as a pleiotropic factor , plays important roles in organ regene2

    ration , organogenesis and morphogenesis , but few reports on protective effect of HGF on injured liver are avail2

    able. The aimof this work was to investigate protective effects of recombinant human HGF (rhHGF) on intoxi2

    cated hepatocytes induced by carbon tetrachloride (CCl4) . The results are as follows : (1) rhHGF (5 ng/ ml)

    could significantly increase the viability of hepatocytes intoxicated by CCl4 (15 mmol/ L) , decrease the leakage

    of intracellular alanine transaminase (ALT) and potassium ions into the culture medium; (2) the combination

    of rhHGF (5 ng/ ml) and epidermal growth factor (EGF) (50 ng/ ml) could increase the viability further , de2

    crease the leakage of intracellular ALT and potassium ions of hepatocytes ; (3) high expression of HGF and its

    receptor/ c2met gene in regenerating liver after partial hepatectomy and CCl4 (50 % , 215 ml/ kg bw) poisoning

    were examined respectively by reverse transcription 2 polymerase chain reaction in rats. The results showed that

    rhHGF could protect hepatocytes against CCl4 through preventing intracellular ALT and potassium ion leakage ,thus suggesting a synergistic effect of rhHGF and EGF on cytoprotection , and high expression of HGF and its

    receptor/ c2met mRNA in the liver treated with PH and CCl4 poisoning. The present study suggests that the ex 2

    pression of HGF and its receptor gene plays an important role in liver regeneration and repair.

    Key words : recombinant human hepatocyte growth factor ; epidermal growth factor ; c 2met ; carbon tetrachlo2

    ride ; primary hepatocyte culture 肝细胞生长因子(hepatocyte growth factor , HGF)

    是由分子量75 kD 的α链和 30 kD 的β链通过二硫

    键连接而成的二聚体[1 ]。HGF在体内广泛分布 , 存

    在于肝、肾、脾、肺、脑、胸腺、甲状腺等处 , 是多

    效性生长因子 , 具有促细胞分裂、促细胞运动和促

    形态形成等作用。HGF受体是原癌基因 c2met 产物 ,分子量为190 kD。我们已初步观察到重组人肝细胞

    生长因子(rhHGF)对原代培养大鼠肝细胞 DNA 合成

    具有显著的刺激作用[2 ]。但是 rhHGF对肝细胞是否

    具有直接的保护作用目前尚未见报道。本工作利用

    原代培养大鼠肝细胞 , 观察 rhHGF对 CCl4 染毒肝细

    胞的保护作用。

    1 材料和方法

    111 主要试剂 rhHGF、重组表皮生长因子

    (EGF) 、胶原酶( Ⅳ型) 、胰岛素、异硫氰酸胍等为

    Sigma 公司产品。 AMV2RTase、TaqDNA聚合酶、低溶

    点琼脂糖(电泳级)为 Promega 公司产品 ......