Page 60 - 2021年18期

Study on the Prevention Mechanism of Anti-tuberculosis Drug-induced Liver Injury with Orazamide
Based on HMGB1-RAGE Signaling Pathway
HE Ling，TANG Jian，PENG Zhongtian（Dept. of Infectious Diseases，the First Affiliated Hospital，Hengyang
Medical School，University of South China，Hunan Hengyang 421001，China）

ABSTRACT OBJECTIVE： To preliminarily investigate the possible mechanism of orazamide to prevent anti-tuberculosis
drug-induced liver injury（ATB-DILI）. METHODS：A total of 60 Kunming mice were randomly divided into blank group，model
group，positive control group [diammonium glycyrrhizinate 60 mg/（kg·d）]，orazamide low-dose，medium-dose and high-dose
groups [80，160，320 mg/（kg·d）]，with 10 mice in each group. Except for blank group，other groups were given isoniazid [75
mg/（kg·d）]+rifampicin [75 mg/（kg·d）] for 14 days intragastrically to induce ATB-DILI model. At the same time，administration
groups were given relevant medicine intragastrically，blank group and model group were given normal saline intragastrically. The
administration volume was 20 mL/（kg·d），once a day，for consecutive 14 days. The general conditions of the mice were observed
and recorded every day，such as growth and development，mental and diet state. After last medication，liver index was calculated，
and HE staining was adopted to observe pathological changes of liver tissue of mice. The positive expression of high mobility group
protein B1 （HMGB1） and NF-κ B in liver tissue were detected by streptavidin biotin-peroxidase complex （SABC） immuno-
histochemistry. The serum levels of liver function indexes in serum，the protein expression of advanced glycation end product
receptor（RAGE）and TNF-α in liver tissue were detected by ELISA. RESULTS：Compared with blank group，the growth and
development of mice in the model group were slow，and their appetite and spirit were poor. The liver index，serum levels of TBIL，
DBIL，ALT，AST，ALP，TBA and γ-GT were increased significantly（P＜0.05）. Structural disorder of liver lobules，degeneration
and necrosis of liver cells and inflammatory cell infiltration were observed. The expression of HMGB1，NF-κB，RAGE and TNF-α
in liver tissue were elevated significantly（P＜0.05）. Compared with model group，the general condition of mice were all improved
to different extents in orazamide low-dose，medium-dose and high-dose groups，positive control group，while liver index and
above serum indexes were all decreased significantly （P＜0.05）. The pathological changes of liver tissue were all improved to
different extents，while the protein expression of HMGB1，NF-κB，RAGE and TNF-α were all decreased significantly（P＜0.05）.
The improvement of above indexes in orazamide high-dose group were all significantly better than orazamide low-dose and
medium-dose groups（P＜0.05）；the levels of ALP and TBA in orazamide high-dose group were significantly lower than positive
control group（P＜0.05）. CONCLUSIONS：Orazamide can prevent ATB-DILI induced by isoniazid combined with rifampicin in
mice，the mechanism of which may be associated with down-regulating the protein expression of HMGB1 and RAGE in liver tissue
and inhibiting the secretion of inflammatory factors.
KEYWORDS Orazamide；Anti-tuberculosis drug-induced liver injury；HMGB1-RAGE signaling pathway；Mice

结核病是由结核分枝杆菌引起的一种多系统传染 联合利福平所致肝细胞变性坏死与氧化应激和炎症反
病，也是人类患病和死亡的主要感染性原因，严重影响 应密切相关，过氧化物酶体增殖物激活受体γ激动剂
患者的生活质量和生命安全，且近年来有抬头趋势，防 15d-PGJ2 可能通过抗氧化及抑制 HMGB1 表达而发挥
控形势十分严峻，是全球性的公共卫生问题之一 [1－2] 。 抗 ATB-DILI 的作用。鉴于 HMGB1 及 RAGE 在肝脏中
[6]
抗结核药物性肝损伤（anti-tuberculosis drug-induced liver 的广泛表达 ，靶向 HMGB1 或 RAGE 在肝病治疗中可
injury，ATB-DILI）是抗结核治疗中最多见的且危害最大 能具有重要的应用前景。奥拉米特（orazamide）又名阿
的不良反应，其可致部分患者不得不中止抗结核治疗， 卡 明（aicamin），是 一 种 临 床 常 用 的 护 肝 药 物 ，对
[7]
既降低了结核病治疗的效果，又增加了耐药结核病的 ATB-DILI具有一定的预防作用 ，但其护肝机制尚未明
[3]
发生风险 。目前，临床一线护肝药物的抗 ATB-DILI 确。本研究通过异烟肼联合利福平复制小鼠ATB-DILI
机制研究有限，亟需寻找切实有效且机制明确的护肝 模型，基于 HMGB1-RAGE 信号通路初步探讨奥拉米特
药物。 预防 ATB-DILI 的作用机制，以期为阐明奥拉米特预防
随着分子生物学和免疫学研究在肝病领域的不断 ATB-DILI的机制提供参考。
深入，有学者发现，高速泳动族蛋白 B1（high mobility 1 材料
group box 1，HMGB1）与晚期糖基化终末产物受体（ad- 1.1 主要仪器
vanced glycation end product receptor，RAGE）可通过多 本研究所用主要仪器包括 7170A 型全自动生化分
种途径来激活促炎因子的相互作用，从而介导肝组织损 析仪（日本Hitachi公司），JY3002型电子天平（上海精密
伤与炎症级联反应的发生 。何雪等 研究表明，异烟肼 科学仪器有限公司），Motic B5 型显微摄像系统、Eco-
[5]
[4]
·2230 · China Pharmacy 2021 Vol. 32 No. 18 中国药房 2021年第32卷第18期