摘要
目的通过建立肝缺血再灌注损伤介导肝纤维化小鼠模型,探讨核因子E2相关因子(Nrf)2/HO-1和Smad通路在肝纤维化早期形成过程中的作用,进一步阐明肝纤维化早期启动的机制。方法将24只C57雄性小鼠分为正常对照组和模型(0、3、6 h)组,每组6只。模型组用血管夹夹闭肝门静脉、入肝中叶和左叶的肝动脉90 min,开夹后再灌注0、3及6 h。检测血浆丙氨酸氨基转移酶(ALT)、乳酸脱氢酶(LDH)和谷胱甘肽过氧化物酶(GSH-Px)含量;测定肝组织超氧化物歧化酶(SOD)、丙二醛(MDA)含量;反转录-聚合酶链反应(RT-PCR)检测肝组织白细胞介素(IL)-6、IL-1β、肿瘤坏死因子(TNF)-α、α-平滑肌肌动蛋白(SMA)、血红素加氧酶(HO)-1、Nrf2、转化生长因子(TGF)-β1含量;Western印迹检测肝组织α-SMA、TGF-β1、Smad3、Smad7的蛋白表达量;苏木素-伊红(HE)染色观察肝组织形态学变化。结果与对照组相比,模型组各组血浆ALT、LDH水平明显升高(P<0.05);血浆GSH-Px水平明显降低(P<0.05);肝组织SOD含量明显降低(P<0.05),而MDA含量明显升高(P<0.05);肝组织IL-6、IL-1b、TNF-α、TGF-β1、α-SMA mRNA的表达量均呈不同程度明显增加(P<0.05),HO-1、Nrf2 mRNA的表达量则呈不同程度明显降低(P<0.05);肝组织TGF-β1、α-SMA、Smad3、Smad7的蛋白表达量呈不同程度明显增加(P<0.05)。HE染色显示,模型组各组肝细胞均有不同程度水肿、脂肪变性、局灶性坏死。结论缺血再灌注损伤介导的早期肝纤维化与Nrf2/HO-1和Smad通路激活都有关,但Smad通路的激活在早期启动中可能不起主要作用。
Objective To explore the role of nuclear factor E2 related factor(Nrf)2/HO-1 and Smad pathway in the early formation of hepatic fibrosis by establishing a mouse model of hepatic ischemia-reperfusion injury,and further clarify the mechanism of early initiation of hepatic fibrosis.Methods Twenty-four C57 male mice were divided into normal control group and model(0,3 and 6 h)groups,with 6 mice in each group.In the model group,the hepatic portal vein,hepatic artery entering the middle and left lobe of the liver were clamped by vascular clamp for 90 minutes,and then reperfused for 0,3 and 6 hours after opening the clip.Plasma alanine aminotransferase(ALT),lactic dehydrogenase(LDH)and glutathion peroxidase(GSH-Px)were detected.The contents of superoxide dismutase(SOD)and malondialdehyde(MDA)in liver tissue were measured.Reverse transcription-polymerase chain reaction(RT-PCR)was used to detect the contents of interleukin(IL)-6,IL-1β,tumor necrosis factor(TNF)-α,α-smooth muscle actin(α-SMA),haem oxygenase(HO)-1,Nrf2 and transforming growth factor(TGF)-βin liver tissue.The expression ofα-SMA,TGF-β1,Smad3 and Smad7 in liver tissue were detected by Western blot.The morphological changes of liver tissue were observed by htoxylin eosin(HE)staining.Results Compared with control group,plasma ALT and LDH levels in model groups were significantly increased(P<0.05),plasma GSH-Px level was significantly decreased(P<0.05),SOD content in liver tissue was significantly decreased(P<0.05),while MDA content was significantly increased(P<0.05),the expressions of IL-6,IL-1β,TNF-α,TGF-β1 andα-SMA mRNA in liver tissue were significantly increased in different degrees(P<0.05),while the expressions of HO-1 and Nrf2 mRNA were significantly decreased in different degrees(P<0.05),the expressions of TGF-β1,α-SMA,Smad3 and Smad7 in liver tissue were significantly increased in different degrees(P<0.05).HE staining showed that liver cells in model groups had edema,steatosis and focal necrosis in different degrees.Conclusions The early hepatic fibrosis mediated by ischemia-reperfusion injury is related to the activation of Nrf2/HO-1 and Smad pathway,but the activation of Smad pathway may not play a major role in the early initiation.
作者
杨洋
宋千黛
赵婧瑶
仝琳鸽
叶圣莹
秦燕
YANG Yang;SONG Qian-Dai;ZHAO Jing-Yao(Department of Physiology and Pathophysiology,School of Basic Medicine,Dali University,Dali 671000,Yunnan,China)
出处
《中国老年学杂志》
CAS
北大核心
2023年第3期598-602,共5页
Chinese Journal of Gerontology
基金
国家自然科学基金(81660111)
大理大学创新团队(ZKLX2020306)。
作者简介
通信作者:秦燕(1973-),女,硕士,教授,硕士生导师,主要从事消化生理与病理研究;第一作者:杨洋(1994-),女,硕士在读,主要从事消化生理与病理研究。
