当归芍药散对肝纤维化大鼠的干预作用及对TLR4通路的影响  

Efficacy of Danggui Shaoyao Powder on liver fibrosis in rats and its effect on TLR4 pathway

作　　者：王雯[1] 胡传国[1] 梁伟林 WANG Wen;HU Chuanguo;LIANG Weilin(The People’s Hospital of Xinjiang Uygur Autonomous Region,Urumqi 830001,Xinjiang,China;Jingxi Medical District,PLA General Hospital,Beijing 100144,China)

机构地区：[1]新疆维吾尔自治区人民医院,新疆乌鲁木齐830001 [2]解放军总医院京西医疗区,北京100144

出　　处：《现代中西医结合杂志》2023年第14期1932-1939,共8页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：自治区卫生健康青年医学科技人才专项科研项目(WJWY-202226);第七批全国老中医药专家学术经验继承工作。

摘　　要：目的研究当归芍药散对四氯化碳诱导的大鼠肝纤维化的影响,并基于Toll样受体4(TLR4)通路探讨其可能作用机制。方法取75只SD大鼠,随机选12只作为正常组,其余大鼠采用腹腔注射四氯化碳-橄榄油法建立肝纤维化模型。将造模成功的大鼠随机分为肝纤维化组,TAK-242组及当归芍药散低、中、高剂量组,每组12只。当归芍药散低、中、高剂量分别给予2.5 g/kg、5 g/kg、10 g/kg的当归芍药散灌胃,TAK-242组腹腔注射3 mg/kg TAK-242,均1次/d,连续13周。使用全自动生化分析仪测定血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、碱性磷酸酶(ALP)水平,ELISA法测定血清白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)水平及肝组织中Ⅳ型胶原(Col-Ⅳ)、Ⅲ型前胶原(PCⅢ)、透明质酸(HA)、层黏连蛋白(LN)水平,Masson染色进行肝组织病理学观察,RT-PCR法测定肝组织中TLR4、髓样分化因子88(MyD88)、肿瘤坏死因子受体相关蛋白6(TRAF6)mRNA表达情况,免疫组化法测定肝组织中TLR4、MyD88、TRAF6蛋白表达平均光密度值,Western blot法测定肝组织中TLR4、MyD88、TRAF6蛋白表达情况。结果与正常组比较,肝纤维化组大鼠血清ALT、AST、ALP、IL-6、IL-1β、TNF-α水平,肝组织中Col-Ⅳ、PCⅢ、HA、LN水平,肝组织中TLR4、MyD88、TRAF6 mRNA及蛋白相对表达量和平均光密度值均显著升高(P均<0.05),肝组织炎症细胞浸润、纤维化增生;与肝纤维化组比较,当归芍药散各组及TAK-242组大鼠血清ALT、AST、ALP、IL-6、IL-1β、TNF-α水平,肝组织中Col-Ⅳ、PCⅢ、HA、LN水平,肝组织中TLR4、MyD88、TRAF6 mRNA及蛋白相对表达量和平均光密度值均显著降低(P均<0.05),肝组织炎症细胞浸润和纤维化增生明显减轻,且各项指标变化与当归芍药散的剂量呈依赖性。结论当归芍药散能够显著改善肝纤维化大鼠肝功能,抑制肝组织炎症,修复肝组织纤维化,其机制可能与调�Objective It is to study the effect of Danggui Shaoyao Powder on carbon tetrachloride-induced liver fibrosis in rats,and to explore its possible mechanism of action based on Toll-like receptor 4(TLR4)pathway.Methods Seventy-five SD rats were taken,in which 12 ones were randomly selected as the normal group,and the rest rats were given carbon tetrachloride(CCl 4)-olive oil by intraperitoneal injection to establish liver fibrosis models.The successfully modeled rats were randomly divided into hepatic fibrosis group,TAK-242 group,and the low,medium,and high dose groups of Danggui Shaoyao Powder,with 12 rats in each group.The low,medium,and high dose groups of Danggui Shaoyao Powder were given Danggui Shaoyao Powder at doses of 2.5 g/kg,5 g/kg,10 g/kg by gavage,respectively,the TAK-242 group was injected intraperitoneally with TAK-242 at a dose of 3 mg/kg,all once daily,continuously treated for 13 weeks.The levels of serum alanine aminotransferase(ALT),aspartate aminotransferase(AST)and alkaline phosphatase(ALP)were determined by automatic biochemical analyzer,the levels of interleukin(IL)-6,IL-1β,tumor necrosis factor-α(TNF-α)in serum and typeⅣcollagen(Col-Ⅳ),typeⅢprocollagen(PCⅢ),hyaluronic acid(HA),laminin(LN)in liver tissue were determined by ELISA,the pathological change of liver tissue was observed by Massonstaining method,the expressions of TLR4,Myd88 and TRAF6 mRNA in liver tissues were detected by RT-PCR,the average integrated optical density(IOD)valuesof Myd88 and TRAF6 in liver tissues were detected by immunohistochemistry,the expression of TLR4,Myd88 and TRAF6 proteins in liver tissues were detected by Western blotting.Results Compared with the normal group,the levels of serum ALT,AST,ALP,IL-6,IL-1βand TNF-α,the levels of Col-Ⅳ,PCⅢ,HA and LN in liver tissues,the relative expression and mean optical density values of TLR4,Myd88 and TRAF6 mRNA and protein in liver tissue of rats in the liver fibrosis groupwere significantly increased(all P<0.05),with obvious hepatic tissue inflammatory cell

关 键 词：当归芍药散 肝纤维化 TOLL样受体4 

分 类 号：R-332[医药卫生]