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

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(CCl_4)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CCl_4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。

Objective This study was aimed to investigate the repair potential of hepatic fibrosis in ratswith 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 maleSD model rats were randomly divided into spontaneous reversal group（n=7）and HPCs transplantation group（n=7） . HPCs transplantation was conducted by intrasplenic injection with5 x106HPCs/0.5 ml. Liver tissues wasstained with hematoxylin-eosin and sirius red, and histopathological changes were observed. The expression of OV6and a-smooth muscle agonists（a-SMA）in liver tissues were detected by immunohistochemical staining, and the a-SMA and tissue inhibitor of matrix metalloproteinases （TIMP） -1 were detected by Western bloting. Serum typeI collagen and albumin levels were measured by ELISA. Results The number of OV6 positive cells in livertissues reached the peak at 8 weeks after intraperitoneal injection of CCl4, and decreased at 12 weeks; the degreeof fibrosis was reduced in HPCs -transplanted group, and the area of collagen was significantly decreased ascompared with in the spontaneous reversal group[ （3.14±0.99）vs. （5.05±0.89）, P〈0.05]; the relative quantification of a-SMA [ （1.22±0.10）vs. （0.31±0.08） ] and TIMP-1 protein [ （0.43±0.09）vs. （0.25±0.032） ] in spontaneousreversal group was much higher as compared with inHPCs transplantation group; serum type I collagenlevels in the spontaneous reversal group was higherthan in the HPCs transplantation group [ （41.33±8.08）ng/ml vs.（31.12 ±10.08） ng/ml, P 〈0.05]. serumalbumin levels in spontaneous reversal group wasremarkably 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 liverfibrosis and improve liver function in rats with liver fibrosis.