Liu Chang, Ding Jing-Xin, Zhou Ying, Yin Zhi-Gang, Luo Hong-Tao, Kong Wei-Tao
School of Pharmacy/Research Center for Application and Development of Medicine and Food Dual-use Resources,Guizhou University of Traditional Chinese Medicine Guiyang 550025, China.
School of Pharmacy/Research Center for Application and Development of Medicine and Food Dual-use Resources,Guizhou University of Traditional Chinese Medicine Guiyang 550025, China Guizhou Engineering Center for Innovative Traditional Chinese Medicine and Ethnic Medicine Guiyang 550025, China.
Zhongguo Zhong Yao Za Zhi. 2021 Sep;46(17):4531-4540. doi: 10.19540/j.cnki.cjcmm.20210624.401.
This study aims to explore underlying mechanism of Lonicerae Japonicae Flos(LJF) in protecting rats against acute alcoholic liver injury(ALI) based on mitogen-activated protein kinase(MAPK) pathway. First, the targets of LJF in preventing ALI were predicted by network pharmacology and the component-target-pathway network was constructed, so that the key targets of LJF components acting on MAPK pathway were screened. Second, male SD rats were randomized into the control(KB) group, model(MX) group, positive(YX) group, and LJF high-(GJ), medium-(ZJ), and low-(DJ) dose groups. Each administration group was given(ig) corresponding drugs for 7 days and KB group and MX group received(ig) equal volume of distilled water every day. Except for KB group, rats were given Chinese spirit(56%, 3 days) for ALI modeling. The levels of aspartate transaminase(AST), alanine transaminase(ALT), interleukin-6(IL6) and tumor necrosis factor-α(TNF-α) in serum and malondialdehyde(MDA), glutathione(GSH), superoxide dismutase(SOD) and glutathione peroxidase(GSH-Px) in liver tissue of rats in each group were detected. Furthermore, we employed quantitative real-time PCR(qRT-PCR) to probe the effects of LJF on the key targets of MAPK pathway in ALI rats. A total of 28 active components of LJF were screened from TCMSP database, and 317 intersected with ALI-related targets. According to Kyoto encyclopedia of genes and genomes(KEGG) pathway enrichment analysis, the 317 targets involved 226 pathways, which were mainly liver disease, inflammation, immunity, apoptosis and other related pathways. According to the MAPK pathway-target-active component network, the key active components of LJF, such as chlorogenic acid, hederagenol, and hyperoside, acted on 25 key targets of MAPK pathway. The results of in vivo experiments showed decreased levels of AST, ALT, and MDA in DJ, ZJ, and GJ groups(P<0.01 or P<0.05), reduced levels of IL6 in DJ and GJ groups(P<0.01 or P<0.05), and improved levels of SOD and GSH in ZJ and GJ groups(P<0.01 or P<0.05). The results of qRT-PCR demonstrated that the expression levels of mitogen-activated protein kinase kinase 4(MAPK2 K4) and mitogen-activated protein kinase 3(MAPK3) were decreased in DJ, ZJ, and GJ groups(P<0.01). The network pharmacology and experimental verification showed that the active components in LJF can reduce the inflammatory factor level and enhance the activities of SOD and GSH-Px by inhibiting the expression of key targets of MAPK pathway, thus alleviating and preventing liver damage caused by alcohol.
本研究旨在基于丝裂原活化蛋白激酶（MAPK）通路探讨金银花保护大鼠急性酒精性肝损伤（ALI）的潜在机制。首先，通过网络药理学预测金银花防治ALI的靶点，构建成分-靶点-通路网络，筛选金银花成分作用于MAPK通路的关键靶点。其次，将雄性SD大鼠随机分为对照组（KB）、模型组（MX）、阳性组（YX）、金银花高剂量组（GJ）、中剂量组（ZJ）和低剂量组（DJ）。各给药组灌胃相应药物7天，KB组和MX组每日灌胃等体积蒸馏水。除KB组外，大鼠给予白酒（56%，3天）建立ALI模型。检测各组大鼠血清中天冬氨酸转氨酶（AST）、丙氨酸转氨酶（ALT）、白细胞介素-6（IL6）和肿瘤坏死因子-α（TNF-α）水平以及肝组织中丙二醛（MDA）、谷胱甘肽（GSH）、超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH-Px）水平。此外，采用定量实时聚合酶链反应（qRT-PCR）探讨金银花对ALI大鼠MAPK通路关键靶点的影响。从中药系统药理学数据库（TCMSP）筛选出金银花28个活性成分，与ALI相关靶点交集317个。根据京都基因与基因组百科全书（KEGG）通路富集分析，317个靶点涉及226条通路，主要为肝脏疾病、炎症、免疫、凋亡等相关通路。根据MAPK通路-靶点-活性成分网络，金银花关键活性成分如绿原酸、常春藤皂苷元、金丝桃苷等作用于MAPK通路25个关键靶点。体内实验结果显示，DJ组、ZJ组和GJ组AST、ALT和MDA水平降低（P<0.01或P<0.05），DJ组和GJ组IL6水平降低（P<0.01或P<0.05），ZJ组和GJ组SOD和GSH水平升高（P<0.01或P<0.05）。qRT-PCR结果表明，DJ组、ZJ组和GJ组丝裂原活化蛋白激酶激酶4（MAPK2K4）和丝裂原活化蛋白激酶3（MAPK3）表达水平降低（P<0.01）。网络药理学及实验验证表明，金银花活性成分可通过抑制MAPK通路关键靶点表达，降低炎症因子水平，增强SOD和GSH-Px活性，从而减轻和预防酒精所致肝损伤。
