黄芪总黄酮对肝硬化大鼠肝组...指标及NF-κB表达的影响_张峰.pdf

526 CHINESE JOURNAL OF ANATOMY Vol.45 No.6 2022 解剖学杂志 2022 年第 45 卷第 6 期doi：10.3969/j.issn.1001-1633.2022.06.007论 著黄芪总黄酮对肝硬化大鼠肝组织病理、代谢指标及NF-B表达的影响张 峰 李 燕 李伟华（济南市人民医院病理科，济南 271100）摘要 目的：目的：探讨黄芪总黄酮对肝硬化大鼠肝组织病理、代谢指标及NF-B表达的影响。方法：方法：将健康大鼠随机分为健康组、肝硬化组、低浓度黄芪总黄酮组（低浓度组）、高浓度黄芪总黄酮组（高浓度组）、美他多辛组。除健康组外，给予其他各组大鼠注射40%四氯化碳花生油建立肝硬化大鼠模型。低浓度组和高浓度组给予不同浓度黄芪总黄酮灌胃；美他多辛组给予美他多辛片灌胃，用药2周。ELISA法检测各组大鼠血清中谷草转氨酶（AST）、谷丙转氨酶（ALT）、总胆红素（TBIL）、肿瘤坏死因子（TNF-）、白细胞介素（IL）6（IL-6）、IL-8、基因结合核因子（NF-B）水平；H-E染色检测大鼠肝组织病理形态；分别采用Realtime PCR和免疫印迹检测各组大鼠肝组织中NF-B、TNF-mRNA及蛋白表达水平。结果：结果：与健康组相比，肝硬化组大鼠血清中AST、ALT、TBIL、TNF-、IL-6、IL-8水平明显升高；与肝硬化组相比，低、高浓度组，美他多辛组上述水平均有所下降；与美他多辛组相比，低、高浓度组降低明显，低、高浓度组间差异无统计学意义。H-E染色可见肝硬化组大鼠肝细胞结构完整、排列整齐；肝硬化组大鼠肝汇管区、小叶间可见大量增生胶原纤维；低、高浓度组，美他多辛组大鼠较肝硬化组肝组织纤维增生明显改善。与肝硬化组相比，低、高浓度组，美他多辛组肝组织中NF-B、NF-B p65、TNF-表达水平明显下降；与美他多辛组相比，低、高浓度组肝组织中NF-B、NF-B p65、TNF-表达水平降低明显，低、高浓度组相比无差异。与健康组比较，肝硬化组可见大量肝细胞凋亡；与肝硬化组相比，低、高浓度组和美他多辛组肝细胞凋亡率均明显降低；与低浓度组相比，高浓度组和美他多辛组大鼠肝细胞凋亡率降低明显，高浓度组与美他多辛组相比无差异。结论：结论：黄芪总黄酮可以改善肝功能水平，抑制TNF-、IL-6、IL-8和NF-B表达，减少肝细胞凋亡，对肝硬化有较好的缓解作用。关键词 黄芪总黄酮；肝硬化；组织病理；代谢指标；基因结合核因子Effects of Astragalus total flavonoids on liver pathology,metabolism indexes and NF-B expression in rats with liver cirrhosisZhang Feng，Li Yan，Li Weihua（Jinan Peoples Hospital，Jinan 271100，China）Abstract Objective：To investigate the effects of Astragalus flavones on liver histomathology，metabolic indexes，and NF-B expression in cirrhosis rats.Methods：Healthy rats were randomly divided into a healthy group，a liver cirrhosis（LC）group，a low concentration of total flavonoids group（LCT group），a high concentration of total flavonoids group（HCT group），and a metadocine（ME）group.Except for the healthy group，other groups were injected with 40%carbon tetrachloride peanut oil to establish liver cirrhosis rat models.The low concentration group and high concentration group were given different concentrations of total flavonoids by gavage.The metadocine group was given metadocine tablets by gavage.The serum levels of aspartate aminotransferase（AST），alanine aminotransferase（ALT），total bilirubin（TBIL），TNF-，IL-6，IL-8 and NF-B were detected by ELISA.The pathological morphology of liver tissue was detected by H-E staining.Realtime PCR and Western blotting were used to detect the mRNA and protein expressions of NF-B and TNF-in liver tissues of rats in each group.Results：Compared with the healthy group，the serum levels of AST，ALT，TBIL，TNF-，IL-6 and IL-8 in LC group were significantly increased.Compared with the LC group，the above levels in the LCT group，the HCT group and the ME group were decreased.Compared with the ME group，the LCT group and the HCT group were significantly decreased，and there was no difference between the LCT group and the HCT group.H-E staining showed that the structure of liver cells in the cirrhosis group was complete and orderly.A large number of proliferative collagen fibers could be seen in hepatic portal area and interlobular area in the LC group.Compared with the LC group，liver tissue fiber proliferation was significantly improved 第 1 作者 E-mail：收稿日期：2021-05-27；修回日期：2022-08-18 527 in the LCT group，the HCT group and the ME group.Compared with the LC group，the expression levels of NF-B，NF-B p65，and TNF-in liver tissues of the LCT group，the HCT group，and the ME group were significantly decreased.Compared with the ME group，the expression levels of NF-B，NF-B p65 and TNF-in liver tissue of the LCT group and the HCT group were significantly decreased，but there was no difference between the LCT group and the HCT group.Compared with the the healthy group，a large number of hepatocyte apoptosis was observed in the LC group.Compared with the LC group，the hepatocyte apoptosis rate in the LCT group，the HCT group and the ME group were significantly decreased.Compared with the LCT group，the hepatocyte apoptosis rate in the HCT group and the ME group was significantly decreased.There was no difference between HCT group and ME group.Conclusions：Astragalus total flavonoids can improve the level of liver function，inhibit the expression of TNF-，IL-6，IL-8 and NF-B，reduce the apoptosis of liver cells，and have a good alleviating mechanism for cirrhosis.Key words astragalus total flavonoids；hepatic cirrhosis；histopathology；metabolic index；NF-B肝硬化（hepatic cirrhosis）是由各种慢性肝病发展到肝纤维化后逐渐演变的过程1。机体肝硬化之后导致各种严重的不良后果2-3。肝硬化的形成是一个循序渐进的过程，最终肝变形变硬形成肝硬化5-6。肝功能受损或病情发展严重到肝硬化时，血液中谷草转氨酶（aspartate transaminase，AST）、谷丙转氨酶（alanine transaminase，ALT）、总胆红素（total bilirubin，TBIL）水平均升高。当肝受到刺激时，机体做出一系列反应，过度应答会出现全身变化，活化的巨噬细胞会释放大量肿瘤坏 死 因 子-（tumor necrosis factor，TNF-）、白细胞介素6（IL-6）等细胞因子，基因结合核因子（nuclear factor-gene binding，NFB）水平也会随之升高。还可诱导炎性因子IL-6的产生，参与肝内炎性损伤过程6。中药在治疗各种疾病方面都起到良好的作用。黄芪味甘性温是临床常用中药材之一，具有固表补气、升阳举陷、利尿消肿等作用7-8。现代药理研究发现，黄芪具有保护腹膜、肝脏、心脑血管等功效，对肺功能及利尿等方面有一定的改善效果9。因此本研究探讨黄芪总黄酮对肝硬化大鼠肝组织病理、代谢指标以及NF-B的影响。1 材料和方法1.1 实验大鼠与试剂1.1 实验大鼠与试剂35只SD大 鼠，购 买 于 南 京 君 科 生 物 工 程公 司，体 质 量190 210 g，分7笼 饲 养，室 温241，湿度45%50%，大鼠活动饮水自由。黄芪总黄酮购于浩翔生物（宝鸡）；四氯化碳购于亿德瑞生物公司（大连）；美他多辛片购买于震元制药公司（浙江）；TRIzol试剂盒购买于百泰克生物公司（无锡）；酶联免疫试剂盒购买于艾博因生物科技（江西）；一抗、二抗购买于西格生物科技（上海）；全自动生化仪购