归黄方调控NLRP3炎症小体介导的Caspase-1/GSDMD通路抑制细胞焦亡治疗慢性非细菌性前列腺炎的机制研究

研究发现慢性非细菌性前列腺炎（CNP）发病与细胞因子失衡密切相关，而NLRP3炎症小体激活Caspase-1/GSDMD通路诱导细胞焦亡，导致IL-1β、IL-18促炎细胞因子大量释放，是引起细胞因子失衡的关键环节。前期研究发现归黄方可下调CNP大鼠炎症细胞因子，阻断NLRP3炎症小体活化的诱导装配及组装激活。据此提出假说，归黄方通过调控NLRP3炎症小体活化，介导Caspase-1/GSDMD通路抑制细胞焦亡治疗CNP。本研究通过动物及细胞实验，构建CNP大鼠模型及巨噬细胞焦亡模型，采用扫描电镜观察细胞焦亡形态，免疫组化、免疫荧光等方法检测细胞焦亡通路NLRP3、Caspase-1、GSDMD等表达；采用Annexin-V/PI染色、TUNEL染色、CCK8在细胞水平检测细胞完整性与活力。阐释归黄方调控NLRP3炎症小体介导细胞焦亡通路治疗CNP的作用机制，为临床应用提供依据。

It has been found that the pathogenesis of chronic nonbacterial prostatitis（CNP） is closely related to the imbalance of cytokines, and the activation of Caspase-1/GSDMD pathway by NLRP3 inflammasome induces pyroptosis, which leads to the release of IL-1βand IL-18 pro-inflammatory cytokines, which is the key link to cause the imbalance of cytokines. Previous studies found that Guihuang Formula could down-regulate inflammatory cytokines in CNP model rats and block the induced assembly and assembly activation of NLRP3 inflammatory bodies. On the basis of this, it is hypothesized that Guihuang Formula regulates the activation of NLRP3 inflammasome and mediates the Caspase-1/GSDMD pathway to inhibit cell death in the treatment of CNP. In this study, CNP model rats and macrophage pyroptosis model were established through animal and cell experiments. The morphology of pyroptosis was observed by scanning electron microscope. The expression of NLRP3, Caspase-1 and GSDMD was detected by immunohistochemistry and immunofluorescence;Cell integrity and viability were detected by Annexin-V/PI staining, TUNEL staining and CCK8 at the cellular level. To explain the mechanism of Guihuang Formula in regulating NLRP3 inflammasome mediated pyroptosis pathway in the treatment of CNP, so as to provide a basis for clinical application.