Cao Kefeng, Jiang Hui, Zhang Lili, Fan Chang, Feng Zhigang, Li Biao, Song Laicheng, Zhang Qun
Department of Laboratory Medicine, Traditional Chinese Medical Hospital of Taihe County, Fuyang, China.
Experimental Center of Clinical Research, The First Affiliated Hospital of Anhui University of Chinese Medicine, Hefei, China.
Comb Chem High Throughput Screen. 2024 Oct 14. doi: 10.2174/0113862073338867240930031800.
Biejiaruangan capsule (BJRGC) is a commonly used traditional Chinese medicine preparation for treating oftreating liver fibrosis (LF), but its specific molecular mechanism is unclear. This study used mass spectrometry, network pharmacology and experimental verification to explore the mechanism of BJRGC against LF.
Ultrahigh-performance liquid chromatography-quadrupole-exactive-orbitrap-mass spectrometry (UHPLC-Q-Exactive-Orbitrap-MS) and network pharmacology were employed to identify and screen the potential components, targets, and signaling pathways of BJRGC against LF. The interaction between the active ingredients and targets was validated using molecular docking. Finally, 5-ethynyl-2'-deoxyuridine (EDU) staining, western blotting (WB), and flow cytometry (FCM) were utilized to further verify the mechanism of BJRGC against LF.
A total of 9 prototype components of BJRGC were identified in serum, most derived from iridoid glycosides and triterpenes in Gardenia jasminoides Ellis and Artemisia scoparia Waldst.et Kit. Network pharmacology predicts that medicine prototype components in serum mostly influence targets such as CDK2, CDK6, and PIK3CG, with the key route being the PI3K/AKT signaling pathway. Molecular docking showed that the major components have good binding properties with key target proteins. The experimental results showed that BJRGC could inhibit the proliferation of HSCs, induce cell cycle arrest and reduce the protein expression of CDK2, CDK6 and PIK3CG.
BJRGC can inhibit the proliferation of HSCs by targeting the protein expression of CDK2, CDK6, and PIK3CG in the PI3K/AKT signaling pathway through its prototype components, such as hyperoside, tumulosic acid, and hederagenin, thereby alleviating LF disease.
鳖甲软肝胶囊（BJRGC）是治疗肝纤维化（LF）常用的中药制剂，但其具体分子机制尚不清楚。本研究采用质谱、网络药理学和实验验证的方法来探索鳖甲软肝胶囊抗肝纤维化的机制。
采用超高效液相色谱-四极杆-高分辨-轨道阱质谱联用仪（UHPLC-Q-Exactive-Orbitrap-MS）和网络药理学来识别和筛选鳖甲软肝胶囊抗肝纤维化的潜在成分、靶点和信号通路。利用分子对接验证活性成分与靶点之间的相互作用。最后，采用5-乙炔基-2'-脱氧尿苷（EDU）染色、蛋白质印迹法（WB）和流式细胞术（FCM）进一步验证鳖甲软肝胶囊抗肝纤维化的机制。
血清中共鉴定出鳖甲软肝胶囊9种原型成分，大多来源于栀子和茵陈中的环烯醚萜苷和三萜类化合物。网络药理学预测血清中的药物原型成分主要影响细胞周期蛋白依赖性激酶2（CDK2）、细胞周期蛋白依赖性激酶6（CDK6）和磷脂酰肌醇-3激酶催化亚基γ（PIK3CG）等靶点，关键途径为磷脂酰肌醇-3激酶/蛋白激酶B（PI3K/AKT）信号通路。分子对接显示主要成分与关键靶蛋白具有良好的结合特性。实验结果表明，鳖甲软肝胶囊可抑制肝星状细胞（HSCs）增殖，诱导细胞周期阻滞，并降低CDK2、CDK6和PIK3CG的蛋白表达。
鳖甲软肝胶囊可通过其原型成分如金丝桃苷、土当归酸和常春藤皂苷元等靶向PI3K/AKT信号通路中CDK2、CDK6和PIK3CG的蛋白表达，抑制肝星状细胞增殖，从而减轻肝纤维化疾病。
