基于网络药理学及分子对接探讨黄芩汤减轻子宫内膜癌化疗不良反应的机制

Mechanism of Huangqintang in Alleviating Side Effect of Chemotherapy for Endometrial Cancer Based on Network Pharmacology and Molecular Docking

目的:基于实验及网络药理学和分子对接技术探讨黄芩汤是否可联合卡铂用于子宫内膜癌化疗及其具体机制。方法:构建裸鼠子宫内膜癌皮下移植瘤模型,当瘤体体积达100 mm3左右时,将24只裸鼠随机分为模型组、黄芩汤组(3.5 g·kg^(-1))、卡铂组(50 mg·kg^(-1))、联合组(黄芩汤3.5 g·kg^(-1)+卡铂50 mg·kg^(-1)),每组6只。模型组给予生理盐水200μL灌胃,每日灌胃2次。每2日测量裸鼠皮下移植瘤的体积及裸鼠的体质量。给药20 d后取裸鼠血液做肾功能及血常规检测,并处死裸鼠取瘤称质量。结合实验结果,通过网络药理学预测黄芩汤联合卡铂可能的作用机制,并通过分子对接预测活性成分的结合部位。结果:黄芩汤组、卡铂组、联合组抑瘤率分别为8.87%、50.33%(P<0.05)、64.66%(P<0.01)。与模型组比较,卡铂组裸鼠体质量、白细胞、红细胞、血红蛋白下降,肌酐及尿酸明显上升(P<0.05);与卡铂组比较,联合组裸鼠体质量、白细胞、血红蛋白明显上升(P<0.05),肌酐及尿酸明显降低(P<0.05)。网络药理学筛选得到黄芩汤与卡铂副作用相关的活性成分114个,作用靶点200个,涉及的核心基因主要是热休克蛋白90AA1(HSP90AA1)、转录因子c-Jun(JUN)、丝裂原活化蛋白激酶(MAPK)等,分子对接表明核心成分汉黄芩素、黄芩素、可与HSP90AA1形成稳定的蛋白复合体,是潜在的活性分子。京都基因与基因组百科全书(KEGG)富集分析表明其可能与调节肿瘤坏死因子(TNF)信号通路、白细胞介素-17(IL-17)信号通路、MAPK信号通路、Toll样受体(TLRs)信号通路相关。结论:黄芩汤未明显增强卡铂抑制子宫内膜癌增殖的作用,但联合用药可缓解卡铂诱导的副作用,其机制可能与中药复杂的调控网络相关。

Objective: To evaluate the utility and mechanism of Huangqintang combined with carboplatin in chemotherapy of endometrial cancer by experiments as well as network pharmacology and molecular docking. Method: The xenograft model of endometrial carcinoma was induced in BALB/c nude mice. When the tumor volume reached about 100 mm3,24 nude mice were randomly assigned into a model group,a Huangqintang group(3.5 g·kg^(-1)),a carboplatin group(50 mg·kg^(-1)),and a combination group(3.5 g·kg^(-1) Huangqintang + 50 mg·kg^(-1) carboplatin),with six mice in each group. The mice in the model group received 200 μL of normal saline by gavage,twice a day. The volume of the tumor and the body weight of the mice were measured every two days. After drug intervention for 20 days,the blood of the mice was collected for renal function and blood routine tests. Then the nude mice were euthanized and the tumor was weighted. In combination with the experimental results,the underlying mechanism of Huangqintang combined carboplatin was predicted through network pharmacology and the binding sites of active components were predicted by molecular docking. Result: The tumor inhibition rates of the Huangqintang group,the carboplatin group, and the combination group were 8.87%,50.33%(P<0.05),and 64.66%(P<0.01),respectively. Compared with the results in the model group,the body weight,leukocyte,erythrocyte,and hemoglobin in the carboplatin group decreased,and creatinine and uric acid increased(P<0.05). Compared with the carboplatin group,the combination group showed increased body weight,leukocyte, and hemoglobin(P<0.05),and decreased creatinine and uric acid(P<0.05). A total of 114 potential active components of Huangqintang involved 200 targets related to the side effects of carboplatin. The core genes involved were mainly heat shock protein 90 AA1(HSP90 AA1),transcription factor c-Jun(JUN),and mitogen-activated protein kinase(MAPK). Molecular docking showed that baicalein and wogonin could form a stable protein complex with HSP90 AA1,serving as potential active molecules. Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis revealed that it might be related to the regulation of tumor necrosis factor(TNF)signaling pathway,interleukin(IL)-17 signaling pathway,MAPK signaling pathway,and toll-like receptor pathway. Conclusion: Huangqintang has no obvious inhibitory effect on endometrial cancer,and the tumor suppression effect is not significantly enhanced after combination with carboplatin,but Huangqintang can alleviate carboplatin-induced side effects. The mechanism may be related to the complex network of Chinese medicine.