背景:在中医理论的指导下,中药(TCM)在发现抗单纯疱疹病毒(HSV)药物方面取得了长足的进步。龙胆泻肝汤(LXD),中华人民共和国药典中记录的制剂,已证明对HSV感染有效。然而,其有效成分和作用机制尚不清楚。
目的:基于网络药理学和实验验证,探讨LXD治疗单纯疱疹病毒感染的有效成分和作用机制。
方法:通过抗病毒试验检测网络分析预测的关键化合物的抗HSV活性。应用高效液相色谱法(HPLC)鉴定LXD水提物的主要成分。进行添加时间测定和感染性抑制可逆性测定以鉴定LCB的潜在抗病毒机制。此外,我们通过体重评估LCB在体内的抗病毒作用,病毒载量,组织学分析,和HSV2感染小鼠模型的生殖器病变评分。
结果:我们的数据表明,某些成分在体外表现出显着的抗HSV1/2活性,包括槲皮素,山奈酚,Wogonin,福蒙素,柚皮苷,黄芩素,异鼠李素,格拉布丁,licochalconeA,棘皮素,奥木精A,异甘草素,pinocembrin,LCB和acacetin.HPLC分析表明LCB是LXD水提物的主要成分。体外实验表明,LCB不仅灭活HSV2颗粒,而且还通过抑制Akt及其下游靶标的磷酸化来抑制HSV2增殖。体内实验证实LCB能显著降低病毒滴度,延迟减肥,减轻阴道感染小鼠模型的阴道组织病理变化。
结论:LCB是LXD的主要组成部分,在体内和体外均具有显着的抗HSV2感染作用。这项研究提供了LXD对HSV感染的治愈功效的额外证据,并为进一步研究中医预防和治疗各种疾病的机制提供了有效的分析方法。
BACKGROUND: Traditional Chinese Medicine (TCM) has made enormous strides recently in the discovery of anti-herpes simplex virus (HSV) drugs under the guidance of TCM theory. Longdan Xiegan Decoction (LXD), a formulation recorded in the Pharmacopoeia of the People\'s Republic of
China, has proved to be effective against HSV infection. However, its effective components and action mechanism remain unclear.
OBJECTIVE: To investigate the effective components and mechanisms of LXD in treating HSV infection based on network pharmacology and experimental validation.
METHODS: The anti-HSV activities of key compounds predicted by network analysis were detected by antiviral tests. High-performance liquid chromatography (HPLC) was applied to identify the main components of the LXD aqueous extract. Time-of-addition assay and infectivity inhibition reversibility assay were conducted to identify the potential antiviral mechanisms of licochalcone B (LCB). Additionally, we assessed the antiviral effect of LCB in vivo by use of body weight, viral load, histological analysis, and scoring of genital lesions in an HSV2-infected mouse model.
RESULTS: Our data demonstrated that some components exhibited significant anti-HSV1/2 activity in vitro, including quercetin, kaempferol, wogonin, formononetin, naringenin, baicalein, isorhamnetin, glabridin, licochalcone A, echinatin, oroxylin A, isoliquiritigenin, pinocembrin, LCB and acacetin. HPLC analysis showed that LCB was the main component of LXD aqueous extract. In vitro experiments revealed that LCB not only inactivated HSV2 particles, but also inhibited HSV2 multiplication through the inhibition of the phosphorylation of Akt and its downstream targets. In vivo experiments confirmed that LCB could significantly reduce viral titer, delay weight loss, and alleviate pathological changes in vaginal tissue in vaginal infection mouse models.
CONCLUSIONS: LCB acted as the main component of LXD, with significant anti-HSV2 infection effects both in vivo and in vitro. This study provides additional evidence of the healing efficacy of LXD against HSV infection and presents an efficient analytical method for further investigation of the mechanisms of TCM in prevention and treatment of various diseases.