抑制剂ICG001联合转化生长因子-β对四氯化碳诱导小鼠肝纤维化的治疗作用及其机制  

Curative effect of ICG001 combined with transforming growth factor-β on carbon tetrachloride-induced liver fibrosis in mice and relevant mechanism

作　　者：任益凡 王晓玲[1,3] 牛艳邦 李雪薇 解军[1,3] 史志勇[1,4] 徐钧[1,5] REN Yi-fan;WANG Xiao-ling;NIU Yan-bang;LI Xue-wei;XIE Jun;SHI ZHI-yong;XU Jun(Shanxi Medical University,Taiyuan 030001,Shanxi Province,China)

机构地区：[1]山西医科大学,山西太原030001 [2]山西医科大学第二医院普外科,山西太原030001 [3]山西医科大学生物化学与分子生物学教研室,山西太原030001 [4]山西省人民医院普外科,山西太原030012 [5]山西医科大学第一医院普外科,山西太原030001

出　　处：《中国生物制品学杂志》2021年第5期547-553,共7页Chinese Journal of Biologicals

基　　金：山西省青年科技研究基金(201901D211516)。

摘　　要：目的探讨抑制剂ICG001联合转化生长因子-β(transforming growth factor-β,TGF-β)对四氯化碳(CCl4)诱导小鼠肝纤维化的治疗作用及其机制。方法将C57BL/6J小鼠随机分为对照组、模型组、ICG001组及ICG001+TGF-β组(IT组)。模型组、ICG001组及IT组腹腔注射20%CCl4(0.1 mL/只),对照组腹腔注射生理盐水(0.2 mL/只),各组每周注射2次,共6周;建模第5周,对照组及模型组每日腹腔注射生理盐水(0.2 mL/只),ICG001组及IT组每日腹腔注射ICG001(0.1 mL/只),同时IT组每2日腹腔再注射TGF-β(0.1 mL/只),各组给药2周。第6周末小鼠称重,眼眶采血,收集肝脏组织。ELISA法检测各组小鼠血清中丙氨酸氨基转移酶(ALT)及天冬氨酸氨基转移酶(AST)含量;肝脏组织切片进行HE及网银染色,显微镜观察病理情况;Q-PCR法检测肝脏组织中炎性因子(IL-6、IL-10和TNFα)及纤维化指标(α-SMA、Col-Ⅰ及FOXO1)mRNA表达水平;Western blot法检测α-SMA、Col-Ⅰ及FOXO1蛋白表达水平。结果与对照组比较,模型组AST及ALT含量均显著升高(P均<0.01),ICG001组较模型组及IT组较ICG001组AST及ALT含量均降低(P均<0.01)。组织学检查提示,ICG001组及IT组均可见病理缓解,且IT组的改善程度较ICG001组明显。Q-PCR检测显示,与模型组比较,ICG001组IL-6、TNFα、α-SMA及Col-ⅠmRNA表达水平均显著降低(P均<0.01),IL-10 mRNA表达水平显著升高(P<0.05),FOXO1 mRNA表达水平差异无统计学意义(P>0.05);与ICG001组比较,IT组IL-6、TNFα、α-SMA及Col-ⅠmRNA表达水平均显著降低(P均<0.01),IL-10及FOXO1 mRNA表达水平均显著升高(P均<0.05)。Western blot分析显示,与模型组比较,ICG001组α-SMA及Col-Ⅰ蛋白表达水平均显著降低(P均<0.01),FOXO1蛋白表达水平差异无统计学意义(P>0.05);与ICG001组比较,IT组α-SMA及Col-Ⅰ蛋白表达水平均显著降低(P均<0.05),FOXO1蛋白表达水平显著升高(P<0.05)。结论与单用ICG001相比,ICG001联合TGF-β对CCl4致小鼠肝纤维化有更好�Objective To investigate the curative effect of inhibitor ICG001 combined with cytokine transforming growth factor-β (TGF-β) on carbon tetrachloride-induced liver fibrosis in mouse model as well as the relevant mechanism.Methods C57BL/6 J mice were randomly divided into control,model,ICG001 and ICG001 combined with TGF-β(IT)groups. The mice in model,ICG001 and IT groups were injected i. p. with 20% carbon tetrachloride,0. 1 mL for each,while those in control group with physiological saline,0. 2 mL for each,2 times per week for 6 weeks. From the 5 th week after modeling,the m ice in ICG001 and IT groups were injected i. p. with ICG001,0. 1 mL for each,while those in control and model groups with physiological saline,0. 2 mL for each,once a day for 2 weeks. Meanwhile,the mice in IT group were injected i. p. with TGF-β,0. 1 mL for each,once 2 days for 2 weeks. At the end of the 6 th week,the mice were weighed,of which the serum samples and liver tissue were collected. The alanine aminotransferase(ALT) and aspartate aminotransferase(AST) levels in sera were determined by ELISA,while the liver tissue was observed for pathological change by microscopy with HE and reticular fiber staining. The transcription levels of mRNAs of inflammatory factors(IL-6,IL-10 and TNF-α)and fibrosis indexes(α-SMA,Col-Ⅰ and FOXO1)in liver tissue were determined by QPCR,while the expression levels of α-SMA,Col-Ⅰand FOXO1 by Western blot. Results The AST and ALT contents in model group were significantly higher than those in control group(each P < 0. 01). However,both the contents were significantly lower in ICG001 group than in model group,and in IT group than in ICG001 group(each P < 0. 01).Histological examination showed pathological remission in both ICG001 and IT groups,which was more obvious in the latter than in the former. Q-PCR showed that,compared with those in model group,the transcription levels of IL-6,TNFα,α-SMA and Col-Ⅰ mRNAs in ICG001 group decreased significantly(each P < 0. 01),while that of IL-10 mRNA increa

关 键 词：抑制剂ICG001 四氯化碳 炎性因子 纤维化指标 蛋白表达 

分 类 号：R657.31[医药卫生—外科学]