摘要
目的建立肝纤维化大鼠模型,观察吡非尼酮(PFD)对肝纤维化大鼠肠道黏膜屏障的影响及可能机制。方法30只大鼠均分为正常对照组、四氯化碳(CCL_(4))组及CCL_(4)+PFD组,除正常对照组腹腔注射等体积生理盐水外、其余大鼠使用50%CCL_(4)橄榄油溶液按0.1 mL/100 g于下腹部腹腔注射建立肝纤维化模型,模型建立成功后CCL_(4)+PFD组按200 mg/kg给予PFD溶液灌胃,其余大鼠按同等体积生理盐水灌胃,连续4周;麻醉处死所有大鼠,取肝脏、脾脏并计算肝脏及脾脏指数,门静脉采血检测各组大鼠血清中丙氨酸转移酶(ALT)、天冬氨酸转移酶(AST)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)及脂多糖(LPS)的含量,肝脏Masson染色和回肠HE染色观察组织形态变化,免疫组织化学染色(IHC)和蛋白质印迹法(Western blot)检测回肠中咬合蛋白-1(claudin-1)、闭合蛋白(occludin)的表达水平。结果成功建立大鼠肝纤维化模型,正常对照组大鼠肝小叶结构清晰、汇管区无纤维沉积,CCL_(4)组大鼠肝小叶结构紊乱、中央静脉周围及汇管区纤维增生明显并可见大量纤维间隔形成,CCL_(4)+PFD组大鼠中央静脉周围及汇管区少量纤维增生、纤维间隔较CCL_(4)组变细变少;CCL_(4)组肝脏及脾脏指数较正常对照组均明显增高(P<0.01),PFD治疗后肝脏及脾脏指数均降低(P<0.05);CCL_(4)组血清中AST、ALT、IL-6,TNF-α及LPS含量均高于正常对照组、PFD治疗后有所降低(P<0.05);CCL_(4)组大鼠回肠上皮黏膜结构较正常对照组紊乱、绒毛变短增宽、部分黏膜上皮出现断裂和脱落及炎性细胞浸润等,PFD治疗后回肠绒毛结构好转、高度(VH)及宽度(VW)较CCL_(4)组改善;CCL_(4)组肠道紧密连接(TJ)蛋白occludin及claudin-1表达均较正常对照组减少(P<0.05)、PFD治疗后其表达均有所增加(P<0.05)。结论PFD在抗肝纤维化的同时能减轻小肠黏膜损伤,对大鼠肠道黏膜屏障具有保护作用,其机制�
Objective To establish a rat model of liver fibrosis and investigate the effect and possible mechanism of pirfenidone(PFD)on the intestinal mucosal barrier in rats with liver fibrosis.Methods Thirty rats were equally divided into normal control,carbon tetrachloride(CCL_(4)),and CCL_(4)+PFD groups.The liver fibrosis model was established by intraperitoneal injection of 50%CCL_(4) olive oil solution at 0.1 mL/100 g in the lower abdomen,while the normal control group received an equal volume of normal saline.After successful modeling,the CCL_(4)+PFD group received 200 mg/kg PFD solution by gavage,while the other groups received an equal volume of normal saline for 4 consecutive weeks.All rats were then anesthetized and sacrificed.Liver and spleen indexes were calculated,and blood was collected from the portal vein to detect serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and lipopolysaccharide(LPS).Masson staining of the liver and HE staining of the ileum were performed to observe histomorphological changes.Immunohistochemical staining(IHC)and Western blot were used to detect the expression levels of claudin-1 and occludin in the ileum.Results The rat model of liver fibrosis was successfully established.The liver lobule structure was clear in the normal control group,with no fibrous deposition in the portal area.In the CCL_(4) group,the liver lobule structure was disordered,with significant fibrous hyperplasia around the central vein and portal area and formation of numerous fibrous septa.The CCL_(4)+PFD group showed less fibrous hyperplasia around the central vein and portal area,with thinner and fewer fibrous septa compared to the CCL_(4) group.The liver and spleen indexes were significantly higher in the CCL_(4) group than in the normal control group(P<0.01)and decreased after PFD treatment(P<0.05).Serum levels of AST,ALT,IL-6,TNF-α,and LPS were higher in the CCL_(4) group than in the normal control group and decreased after PFD t
作者
陈沁
程芝梅
何慧洲
张帅
周石
CHEN Qin;CHENG Zhimei;HE Huizhou;ZHANG Shuai;ZHOU Shi(Department of Intervention,the Affiliated Hospital of Guizhou Medical University,Guiyang 550000,Guizhou,China;School of Medical Imaging,Guizhou Medical University,Guiyang 550000,Guizhou,China)
出处
《贵州医科大学学报》
CAS
2024年第6期803-810,共8页
Journal of Guizhou Medical University
基金
国家自然科学基金(81760325)。
关键词
吡非尼酮
肝纤维化
肠黏膜屏障
紧密连接
脂多糖
pirfenidone
liver fibrosis
intestinal mucosal barrier
tight junction
lipopolysaccharide
