Table 2_Curdione combined with borneol treats bacterial mixed HPV infection by regulating the crosstalk among immune cells.docx

BackgroundHuman papillomavirus (HPV) infection is a worldwide reproductive system disease. Baofukang suppository, a traditional herbal preparation that includes curdione and borneol, has been reported to treat bacterial vaginosis (BV) and HPV infection in China. However, the therapeutic mechanism is still unknown. This study aims to explore the molecular mechanisms of curdione and borneol in treating HPV infection.
MethodsWe conducted a retrospective cohort analysis of medical records from a single-center study involving 205 HPV patients, focusing on the correlation between HPV clearance and co-infection with other pathogens, confirming the efficacy of Baofukang suppository. Bioinformatics and network pharmacology approaches were employed to identify therapeutic targets of Baofukang suppository for BV/HPV co-infections. qRT-PCR, Western blot, immunofluorescence staining, and flow cytometry were utilized to validate the therapeutic targets of curdione and borneol, along with the associated immune molecular changes. Finally, the molecular mechanisms and therapeutic efficacy of curdione and borneol were confirmed in vivo using an LPS/TC-1 cervical orthotopic injection model.
ResultsCurdione and borneol selectively inhibit the secretion of interleukin-6 (IL-6) and interleukin-1β (IL-1β) by macrophages. The reduction in IL-6 and IL-1β levels effectively inhibits the expression of CD274 (Programmed death ligand 1, PD-L1) in infected epithelial cells by inhibiting STAT3 phosphorylation, thereby suppressing their immune evasion capabilities. Furthermore, curdione and borneol enhance the expression of tumor necrosis factor α (TNF-α) and caspase 1 (CASP1) in macrophages, as well as the expression of interleukin 12 (IL-12) and interleukin 23 (IL-23) in dendritic cells (DCs). The expression of these inflammatory factors effectively promotes the migration and differentiation of T cells to the site of infection, completing the clearance of infected epithelial cells.
ConclusionThe main components of Baofukang suppository, curdione and borneol, inhibit the progression of HPV infection and the occurrence of cervical cancer by modulating the communication between innate and adaptive immunity, promoting the recruitment and recognition of CD8+ T cells to eliminate HPV-infected epithelial cells.

背景 人乳头瘤病毒（Human papillomavirus, HPV）感染是一种全球性生殖系统疾病。保妇康栓作为包含莪术醇与冰片的传统中药制剂，在中国已被应用于治疗细菌性阴道病（bacterial vaginosis, BV）与HPV感染，但其具体治疗机制仍未明确。本研究旨在探究莪术醇与冰片治疗HPV感染的分子机制。
方法 本研究针对单中心队列的205例HPV感染者的病历资料开展回顾性队列分析，重点探讨HPV清除与其他病原体合并感染之间的相关性，以验证保妇康栓的临床疗效。采用生物信息学与网络药理学方法，筛选保妇康栓用于治疗BV/HPV合并感染的潜在治疗靶点。通过实时荧光定量PCR（qRT-PCR）、蛋白质免疫印迹（Western blot）、免疫荧光染色及流式细胞术，验证莪术醇与冰片的治疗靶点及其相关免疫分子变化。最后，采用脂多糖（lipopolysaccharide, LPS）/TC-1宫颈原位注射模型，在体内验证莪术醇与冰片的分子机制与治疗效果。
结果 莪术醇与冰片可选择性抑制巨噬细胞分泌白细胞介素-6（interleukin-6, IL-6）与白细胞介素-1β（interleukin-1β, IL-1β）。通过抑制信号转导与转录激活因子3（signal transducer and activator of transcription 3, STAT3）的磷酸化，IL-6与IL-1β水平的降低可有效抑制感染上皮细胞中CD274（程序性死亡配体1, Programmed death ligand 1, PD-L1）的表达，从而削弱其免疫逃逸能力。此外，莪术醇与冰片可促进巨噬细胞中肿瘤坏死因子α（tumor necrosis factor α, TNF-α）与半胱天冬酶1（caspase 1, CASP1）的表达，以及树突状细胞（dendritic cells, DCs）中白细胞介素12（interleukin 12, IL-12）与白细胞介素23（interleukin 23, IL-23）的表达。上述炎性因子的表达可有效促进T细胞向感染部位迁移与分化，最终完成对感染上皮细胞的清除。
结论 保妇康栓的主要活性成分莪术醇与冰片，可通过调控固有免疫与适应性免疫之间的信号交流，促进CD8+ T细胞的招募与识别以清除HPV感染的上皮细胞，从而抑制HPV感染进展与宫颈癌发生。