摘要
目的:研究壮方柔肝化纤颗粒对CCl_(4)复合因素诱导肝纤维化大鼠模型的干预效应及其对α-SMA、E-Cadherin蛋白表达的影响。方法:选取48只雄性Wistar大鼠随机分成空白组、病理模型组与柔肝化纤颗粒组,每组均为16只。病理模型组与柔肝化纤颗粒组大鼠皮下注射40%四氯化碳(CCl_(4))油剂联合花生油复合因素建立大鼠肝纤维化动物模型,第5周起给予柔肝化纤颗粒灌胃给药,每周灌胃3次,连续8周,8周后检测肝功能转氨酶指标(ALT、AST),通过HE染色、Masson染色、免疫组化等方法,对比分析各组大鼠肝组织的病理学形态改变,采用酶联免疫分析(ELISA)检测大鼠氧化应激氧化物歧化酶(SOD)与丙二醛(MDA)的表达,并采用RT-PCR、Western Blot等方法检测α-平滑肌肌动蛋白(α-SMA)、E-钙粘蛋白(E-Cadherin)的表达。结果:与空白组比较,病理模型组大鼠血清的ALT、AST水平明显升高,柔肝化纤颗粒组大鼠血清ALT、AST水平均明显下降(P均<0.001)。病理模型组大鼠肝组织内的小叶结构已残缺,出现众多肝脏纤维组织异常增生,增生所形成的胶原纤维汇合成密集的纤维间隔,从而变成假小叶结构,假小叶结构内部出现大量形态各异、大小不均的脂肪变性的肝细胞,炎症细胞弥漫性浸润显著。与病理模型组相比,柔肝化纤颗粒组肝小叶结构被破坏的程度显著减轻,有不同程度增生的胶原纤维组织集中在中央静脉至汇管区范围,纤维间隔相对疏松,有少量炎症细胞浸润,浸润程度较病理模型组减轻。与病理模型组比较,柔肝化纤颗粒组SOD水平较病理模型组与空白组明显升高(P<0.001),柔肝化纤颗粒组MDA水平较病理模型组明显下降(P<0.001),柔肝化纤颗粒组α-SMA的表达水平明显降低(P<0.001),E-Cadherin的表达水平均明显升高(P<0.001)。结论:柔肝化纤颗粒能有效抑制肝纤维组织增生,降低CCl_(4)联合花生油复合溶液联合诱导肝纤维化大鼠血清的ALT、AST水平,其抗肝纤维化机制可能是通过保护肝功能、降低氧化应激水平、下调α-SMA蛋白的表达、上调E-cadherin蛋白的表达、抑制上皮细胞-间充质转化(epithelial mesenchymal transformation,EMT)从而对CCl_(4)复合因素诱导肝纤维大鼠化发挥治疗作用。
Objective:To study the intervention effect of Zhuangfang Soft Liver and Fibrous Granules on CCl_(4)compound factor-induced liver fibrosis rat model and its effect onα-SMA and E-Cadherin protein expression.Methods:Forty-eight male Wistar rats were randomly divided into a blank group,a pathological model group and a group of 16 rats in each group.The rats in the pathological model group and the pellet group were injected subcutaneously with 40%carbon tetrachloride(CCl_(4))oil combined with peanut oil to establish an animal model of rat liver fibrosis,and the pellets were administered by gavage three times a week for 8 weeks from the 5th week.The expression of oxidative stress oxidase dismutase(SOD)and malondialdehyde(MDA)was detected by enzyme-linked immunoassay(ELISA),and the expression ofα-smooth muscle actin(α-SMA)and E-calmodulin(E-Cadherin)was detected by RT-PCR and Western Blot.expression.Results:Compared with the blank group,the serum ALT and AST levels of the rats in the pathological model group were significantly increased,and the serum ALT and AST levels of the rats in the soft liver and fibrous pellet group were significantly decreased(P<0.001).In the pathological model group,the lobular structure of liver tissue was mutilated and numerous liver fibrous tissues were abnormally proliferated,and the collagen fibers formed by the proliferation were confluent into dense fibrous intervals,thus turning into pseudolobular structure,and a large number of fatty degenerated hepatocytes of different shapes and sizes appeared inside the pseudolobular structure,and the diffuse infiltration of inflammatory cells was significant.Compared with the pathological model group,the degree of destruction of hepatic lobular structure was significantly reduced in the LBP group,and there were different degrees of proliferation of collagen fibrous tissue concentrated in the area from the central vein to the confluent area,and the fibrous septum was relatively loose,with a small amount of inflammatory cell infiltration,and the degree of infiltration was reduced compared with the pathological model group.Compared with the pathological model group,the SOD level was significantly higher in the group of liver softener and fibrous pellets compared with the pathological model group and the blank group(P<0.001),the MDA level was significantly lower in the group of liver softener and fibrous pellets compared with the pathological model group(P<0.001),the expression level ofα-SMA was significantly lower in the group of liver softener and fibrous pellets(P<0.001),and the expression level of E-Cadherin was significantly higher in the group of liver softener and fibrous pellets(P<0.001).Conclusion:Soft Liver Fibro Pellets can effectively inhibit liver fibrous tissue proliferation and reduce the serum ALT and AST levels in rats with CCl_(4)combined with peanut oil compound solution induced liver fibrosis,and its anti-liver fibrosis mechanism may be through protecting liver function,reducing the level of oxidative stress,down-regulating the expression ofα-SMA protein,up-regulating the expression of E-cadherin protein,and inhibiting epithelial-mesenchymal transition epithelial mesenchymal transformation(EMT)and thus exerting therapeutic effects on CCl_(4)compound-induced hepatic fibrosis in rats.
作者
张文富
吴姗姗
戴铭
吕建林
黄晶晶
李晓龙
王振常
ZHANG Wen-fu;WU Shan-shan;DAI Ming;LV Jian-lin;HUANG Jing-jing;LI Xiao-long;WANG Zhen-chang(The First Affiliated Hospital of Guangxi University of Traditional Chinese Medicine,Nanning 530023,China;Guangxi Key Laboratory of Integrative Translational Medicine for High Incidence Infectious Diseases,Nanning 530299,China;Guangxi International Zhuang Medicine Hospital,Nanning 530201,China;Guangxi University of Traditional Chinese Medicine,Nanning 530299,China;Guangxi Key Laboratory of Basic Research in Traditional Chinese Medicine,Nanning 530299,China;Guangxi Medical University,Nanning 530021,China)
出处
《海南医学院学报》
CAS
2023年第5期321-330,共10页
Journal of Hainan Medical University
基金
国家自然科学基金项目(81960910、81660745、81860839、8176150106)
广西自然科学基金项目(2020GXNSFDA297021、2020GXNSFAA238020)
戴铭广西名中医工作室建设项目(2022004-005-009)。
