肝脏前体细胞移植对CCl4诱导的肝纤维化自发逆转大鼠肝组织学的影响*

doi:10.3969/j.issn.1672-5069.2018.03.015

实用肝脏病杂志 ›› 2018, Vol. 21 ›› Issue (3): 376-379.doi: 10.3969/j.issn.1672-5069.2018.03.015

肝脏前体细胞移植对CCl4诱导的肝纤维化自发逆转大鼠肝组织学的影响*

陈辉, 任万雷, 杨爱婷

100043 北京市首都医科大学附属北京朝阳医院西院消化科（陈辉）; 附属北京友谊医院科研实验中心/北京市临床医学研究所（杨爱婷）; 青岛市中心医院中医科（任万雷）

收稿日期:2017-03-13 出版日期:2018-05-10 发布日期:2018-05-25
通讯作者: 杨爱婷,E-mail：yangaiting168@126.com
作者简介:陈辉,男,33岁,硕士研究生,住院医师。主要从事消化内镜诊断与治疗学研究。E-mail：chenhui_0516@126.com
基金资助:
*国家自然科学基金资助项目（编号：81500456）

Hepatic progenitor cell transplantation accelerates spontaneous reverse of hepatic fibrosis in rats with carbon tetrachloride-induced liver fibrosis

Chen Hui, Ren Wanlei, Yang Aiting

Department of Gastroenterology,Chaoyang Hospital,Affiliated to Capital Medical University,Beijing 100043,China

Received:2017-03-13 Online:2018-05-10 Published:2018-05-25

摘要/Abstract

摘要： 目的研究肝脏前体细胞（HPCs）移植对纤维化自发逆转大鼠肝脏组织学的影响。方法采用四氯化碳（CCl4）腹腔注射制备大鼠肝纤维化模型,取14只模型大鼠,随机分为自发逆转组7只和HPCs移植组7只,后者采用经脾内注射HPCs。取肝组织行苏木精-伊红和天狼猩红染色,观察组织病理学变化;采用免疫组织化学法检测肝组织OV6和α-平滑肌肌动蛋白（α-SMA）表达;采用Western blot法测定α-SMA、基质金属蛋白酶组织抑制剂（TIMP）-1蛋白表达;采用ELISA法测定血清I型胶原和白蛋白水平。结果在CCl4腹腔注射8 w后动物肝组织OV6阳性细胞数量达到高峰,12 w时开始下降;与自发逆转组比,HPCs移植组肝组织纤维化和炎症程度减轻,胶原面积显著降低【（3.14±0.99）对（5.05±0.89）,P<0.05】;自发逆转组肝组织α-SMA和TIMP-1蛋白相对表达量分别为（1.22±0.10）和（0.43±0.09）,均显著高于HPCs移植组的【（0.31±0.08）和（0.25±0.032）,P<0.05】;自发逆转组血清I型胶原水平为（41.33±8.08） ng/ml,显著高于HPCs移植组的【（31.12±10.08）ng/ml,P<0.05】,自发逆转组白蛋白为（33.18±4.69） g/L,显著低于HPCs移植组的【（48.30±2.15）g/L,P<0.05】。结论肝脏前体细胞移植能加速肝纤维化的逆转,改善肝功能。

关键词: 肝纤维化, 肝脏前体细胞, 细胞移植, 大鼠

Abstract: Objective T his study was aimed to investigate the repair potential of hepatic fibrosis in rats with carbon tetrachloride （CCl4）-induced liver fibrosis after hepatic progenitor cell （HPC） transplantation. Methods CCl4 was intraperitoneally injected for 4 weeks,8 weeks and 12 weeks to establish the model. Immunohistochemical staining was used to detect the expression of OV6-positive HPCs in liver tissues. 14 male SD model rats were randomly divided into spontaneous reversal group （n=7） and HPCs transplantation group （n=7）. HPCs transplantation was conducted by intrasplenic injection with 5×106 HPCs/0.5 ml. Liver tissues was stained with hematoxylin-eosin and sirius red,and histopathological changes were observed. The expression of OV6 and α-smooth muscle agonists（α-SMA） in liver tissues were detected by immunohistochemical staining,and the α-SMA and tissue inhibitor of matrix metalloproteinases （TIMP）-1 were detected by Western bloting. Serum type I collagen and albumin levels were measured by ELISA. Results The number of OV6 positive cells in liver tissues reached the peak at 8 weeks after intraperitoneal injection of CCl4,and decreased at 12 weeks;the degree of fibrosis was reduced in HPCs-transplanted group,and the area of collagen was significantly decreased as compared with in the spontaneous reversal group[（3.14±0.99） vs.（5.05±0.89）,P<0.05];the relative quantification of α-SMA [（1.22±0.10） vs. （0.31±0.08）] and TIMP-1 protein [（0.43±0.09） vs.（0.25±0.032）] in spontaneous reversal group was much higher as compared with in HPCs transplantation group;serum type I collagen levels in the spontaneous reversal group was higher than in the HPCs transplantation group[（41.33±8.08） ng/ml vs.（31.12±10.08） ng/ml,P<0.05]. serum albumin levels in spontaneous reversal group was remarkably lower than that in the HPCs transplanta-tion group [（33.18±4.69） g/L vs. （48.30±2.15） g/L，P<0.05]. Conclusion Hepatic progenitor cell transplantation have the potential to accelerate the reverse of liver fibrosis and improve liver function in rats with liver fibrosis.

Key words: Liver fibrosis, Hepatic progenitor cells, Cell transplantation, Rats

陈辉, 任万雷, 杨爱婷. 肝脏前体细胞移植对CCl4诱导的肝纤维化自发逆转大鼠肝组织学的影响*[J]. 实用肝脏病杂志, 2018, 21(3): 376-379.

Chen Hui, Ren Wanlei, Yang Aiting. Hepatic progenitor cell transplantation accelerates spontaneous reverse of hepatic fibrosis in rats with carbon tetrachloride-induced liver fibrosis[J]. JOURNAL OF PRACTICAL HEPATOLOGY, 2018, 21(3): 376-379.

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肝脏前体细胞移植对CCl4诱导的肝纤维化自发逆转大鼠肝组织学的影响*

Hepatic progenitor cell transplantation accelerates spontaneous reverse of hepatic fibrosis in rats with carbon tetrachloride-induced liver fibrosis

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