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荜茇提取物对CCl4诱导大鼠肝纤维化的影响

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目的 探究荜茇提取物对四氯化碳(CCl4)诱导大鼠肝纤维化的作用机制.方法 将60只SD大鼠随机分成空白组、模型组、荜茇提取物低剂量组(16.6mg/kg)、荜茇提取物中剂量组(33.3mg/kg)和荜茇提取物高剂量组(66.7mg/kg),每组各12只.除空白组外,其余各组均给予皮下注射50%CCl4-菜籽油溶液干预8周,建立肝纤维化模型.造模的同时给予模型组灌胃0.9%氯化钠溶液,荜茇提取物低、中、高剂量组连续灌胃治疗8周.实验结束后,测定大鼠血清谷丙转氨酶(alanine amin-otransferase,ALT)、谷草转氨酶(aspartate aminotransferase,AST)、谷氨酰基转移酶(γ-glutamyltransferase,γ-GT),肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6);苏木精-伊红染色(hematoxylin-eosin stai-ning,HE)、Masson染色观察大鼠肝组织病理变化.提取RNA并采用qPCR技术检测大鼠肝组织中Ⅰ型胶原蛋白(Collagen Ⅰ)、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、TNF-α mRNA表达水平;提取蛋白采用Western blot法检测大鼠肝组织中Collagen Ⅰ、α-SMA、TNF-α蛋白表达水平.结果 HE染色和Masson染色结果可观察到模型组大鼠肝脏纤维增生明显,大鼠血清中ALT、AST、γ-GT升高,血清中炎性介质TNF-α和IL-6分泌增多,Collagen Ⅰ、α-SMA和TNF-α mRNA和蛋白表达增高,荜茇提取物可明显改善肝纤维化程度,血清中ALT、AST、γ-GT下降,血清中炎性介质TNF-α和IL-6下降,肝组织中Collagen Ⅰ、α-SMA、TNF-α mRNA和蛋白水平明显下降.结论 荜茇提取物对CCl4诱导的大鼠肝纤维化有明显治疗作用,可通过抑制炎性细胞因子TNF-α和IL-6表达,可能通过抑制肝星状细胞激活,从而减少Collagen Ⅰ和α-SMA合成发挥抗纤维化的药效.
Effect of Piper Longum Extract on CCl4 Induced Liver Fibrosis in Rats
Objective To explore the mechanism of piper longum extract on liver fibrosis induced by carbon tetrachloride(CCl4)in rats.Methods Sixty SD rats were randomly divided into blank group,model group,low dose group(16.6mg/kg),middle dose group(33.3mg/kg)and high dose group(66.7mg/kg)of piper longum extract,with 12 rats in each group.Except the blank group,the other groups were subcutaneously injected with 50%CCl4 rapeseed oil solution for 8 weeks to establish liver fibrosis models.At the same time of modeling,the model group was given normal saline by gavage,and the low,middle and high dose groups of piper longum extract were given gavage for 8 weeks.After the experiment,serum alanine transaminase(ALT),aspartate aminotransferase(AST)and γ-glutamyl-transferase(γ-GT),tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)were measured;the pathological changes of liver tissue in rats was observed by hematoxylin eosin staining and Masson's trichrome staining.The tissue total RNA was extracted and qPCR method was used to detect the mRNA expression levels of Collagen Ⅰ,α-smooth muscle actin(α-SMA)and TNF-α.Tissue protein was extracted and Western blot method was used to detect the protein expression levels of Collagen Ⅰ,α-SMA and TNF-α.Results The results of HE staining and Masson staining showed that the liver fibrosis of rats in the model group was obvious,ALT,AST and γ-GT were increased,serum inflammatory mediator TNF-α and IL-6 secretion was increased.The mRNA and protein expression levels of Collagen Ⅰ,α-SMA and TNF-α in the model group were significantly increased.Piper longum extract can significantly improve the degree of liver fibrosis.ALT,AST,y-GT were decreased,and serum inflammatory mediator TNF-α and IL-6 were decreased,the mRNA and protein expression levels of Collagen Ⅰ,α-SMA,TNF-α in liver tissue were significantly decreased.Conclusion Piper longum extract has obvious therapeutic effect on CCl4 induced liver fibrosis in rats,which can be achieved by inhibiting the inflammatory factor TNF-α and IL-6 expression,and possibly by inhibiting the activation of hepatic stellate cell,thereby reducing Collagen Ⅰ andα-SMA synthesis exerts anti fibrosis effect.

Piper longum extractCarbon tetrachlorideLiver fibrosisTumor necrosis factor-αRats

郭梓琪、赵丽、王如梦、吴迪、杨勇、杨宏新

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010110 呼和浩特,内蒙古医科大学细胞生物学教研室

010017 呼和浩特,内蒙古自治区人民医院腹部肿瘤外科

荜茇提取物 四氯化碳 肝纤维化 肿瘤坏死因子-α 大鼠

国家自然科学基金内蒙古医科大学重点项目内蒙古医科大学"三位一体"大学生创新创业训练计划培育项目

81960772YKD2022ZD016SWYT2021030

2024

医学研究杂志
中国医学科学院

医学研究杂志

CSTPCD
影响因子:0.702
ISSN:1673-548X
年,卷(期):2024.53(5)