HMGB1介導(dǎo)的炎癥通路在雷公藤致大鼠肝損傷中的作用研究

董捷鳴 章從恩 于小紅 吳昊 馬致潔 趙奎君

中圖分類(lèi)號(hào) R965.1 文獻(xiàn)標(biāo)志碼 A 文章編號(hào) 1001-0408（2018）05-0633-04

DOI 10.6039/j.issn.1001-0408.2018.05.14

摘 要 目的：研究高遷移率族蛋白B1（HMGB1）介導(dǎo)的炎癥通路HMGB1-Toll樣受體4（TLR4）/核轉(zhuǎn)錄因子κB（NF-κB）在雷公藤致大鼠肝損傷中的作用，為闡明雷公藤致肝損傷的作用機(jī)制提供參考。方法：將24只SD大鼠隨機(jī)分為空白組（生理鹽水，灌胃）、雷公藤組（以生藥計(jì)16 g/kg，灌胃）和中和劑組（腹腔注射100 mg/kg甘草酸銨溶液3 h后再灌胃以生藥計(jì)16 g/kg的雷公藤），每組8只，各組大鼠均連續(xù)給藥3周。每周給藥后均檢測(cè)大鼠血清中天冬氨酸轉(zhuǎn)氨酶（AST）和丙氨酸轉(zhuǎn)氨酶（ALT）水平；給藥結(jié)束后，采用酶聯(lián)免疫吸附法檢測(cè)大鼠血清中HMGB1、白細(xì)胞介素1β（IL-1β）、IL-2、腫瘤壞死因子α（TNF-α）水平，Western blot法檢測(cè)大鼠肝組織中HMGB1、NF-κB p65、TLR4蛋白表達(dá)，蘇木精-伊紅染色后觀察大鼠肝組織病理學(xué)改變。結(jié)果：給藥3周后，雷公藤組大鼠血清中AST、ALT、HMGB1、IL-1β、IL-2、TNF-α水平以及肝組織中HMGB1、NF-κB p65、TLR4蛋白表達(dá)水平均顯著高于空白組和中和劑組（P<0.05或P<0.01）。雷公藤組大鼠肝組織中央靜脈周?chē)渭?xì)胞水腫，部分肝細(xì)胞漿內(nèi)可見(jiàn)圓形空泡；中和劑組大鼠僅少量細(xì)胞內(nèi)可見(jiàn)大小不一的空泡。結(jié)論：雷公藤致大鼠肝損傷的機(jī)制可能與其激活了HMGB1-TLR4/NF-κB炎癥通路有關(guān)。

關(guān)鍵詞 雷公藤；肝損傷；高遷移率族蛋白1；Toll樣受體4；核轉(zhuǎn)錄因子κB；大鼠

ABSTRACT OBJECTIVE： To study the effects of high-mobility group box 1 protein （HMGB1）-mediated inflammatory pathway HMGB1-Toll like receptor 4 （TLR4）/nuclear transcription factor κB（NF-κB）on liver injury of rats induced by Tripterygium wilfordii， and to provide reference for clarify the mechanism of liver injury induced by T. wilfordii. METHODS： Totally 24 SD rats were randomly divided into blank group （normal saline， i.g.）， T. wilfordii group （16 g/kg by crude drug， i.g.） and neutralizer group （16 g/kg T. wilfordii crude drug i.g. after i.p injection of 100 mg/kg Ammonium glycyrrhizinate solution 3 h）， with 8 rats in each group. All rats were treated for consecutive 3 weeks. The serum levels of AST and ALT in rats were detected every week. After the end of medication， the serum levels of HMGB1， IL-1β， IL-2 and TNF-α were detected by ELISA method； the protein expression of HMGB1， NF-κB p65 and TLR4 in liver tissue of rats were detected by Western blot assay. The pathological changes of liver tissue in rats were measured with HE staining method. RESULTS： After 3 weeks of treatment， the serum levels of AST， ALT， HMGB1， IL-1β， IL-2 and TNF-α in rats， the protein expression of HMGB1， NF-κB p65 and TLR4 in liver tissue of rats in T. wilfordii group were significantly higher than blank group and neutralizer group （P<0.05 or P<0.01）. Hepatocyte edema was found around the central vein of the liver， and circular vacuoles were seen in some hepatic cytoplasm in T. wilfordii group； only varying size of vacuoles were found in a small number of cells in neutralizer group. CONCLUSIONS： T. wilfordii induced liver injury may be associated with the activation of HMGB1-TLR4/NF-κB inflammation pathway.

KEYWORDS Tripterygium wilfordii； Liver injury； High-mobility group box 1 protein； Toll like receptor 4； Nuclear transcription factor κB； Rats

雷公藤為衛(wèi)矛科植物雷公藤Tripterygium wilfordii Hook.F.的干燥根莖，首載于《神農(nóng)本草經(jīng)》[1]。雷公藤在治療自身免疫性疾病（如類(lèi)風(fēng)濕性關(guān)節(jié)炎、系統(tǒng)性紅斑狼瘡）上有顯著且獨(dú)特的療效[2-3]，但其對(duì)肝、腎以及生殖器官具有一定毒性，在傳統(tǒng)中藥中毒性名列前位[4-5]，從而制約了其臨床應(yīng)用。目前研究認(rèn)為，雷公藤甲素、雷公藤紅素、雷公藤生物堿等是雷公藤致毒的主要成分[6-7]。筆者前期對(duì)雷公藤肝毒性的物質(zhì)基礎(chǔ)也進(jìn)行了相關(guān)研究[8-9]，證實(shí)了雷公藤甲素、雷公藤紅素可能是雷公藤致肝損傷的關(guān)鍵成分，且發(fā)現(xiàn)雷公藤致肝損傷后炎癥因子[白細(xì)胞介素1（IL-1）、IL-6、腫瘤壞死因子α（TNF-α）]水平升高。……