中药槐耳通过抑制caspase-3/GSDME依赖的细胞焦亡途径防治系膜增生性肾小球肾炎

Mesangial proliferative glomerulonephritis（MsPGN） is a disease associated with immune-inflammatory.Persistent immune-inflammatory and subsequent mesangial proliferation will lead to glomerulosclerosis and further end-stage renal disease which causing great burden for social and family.How to regulate immune-inflammatory response so that Inhibit mesangial proliferation is an essential therapeutic strategy for MsPGN. Pyroptosis is a gasdermin-mediated cellular process of inflammatory necrosis that leads to the release of cellular contents and subsequently activate a strong inflammatory response. Recent studies suggest that GSDME could be activated by caspase-3 that lead to pryroptosis. Huaier, as a adjuvant drug for malignant tumors, has immunomodulatory and antitumor effects. It could also inhibit the abnormal proliferation of rat renal mesangial cells. Therefore we want to study the intervention of Huaier on the anti-Thy-1 nephritis rat model that minic human MsPGN steadily.Different methylation genes of pyroptosis and immune-inflammatory during the course of the disease are detected by MeDIP-seq and Bioinformatics. Rat mesangial cell are incubated in vitro by RNAi and plasmid transfection. The mechanism of Huaier on MsPGN that focus on the mechanism of immuno-inflammatory regulation of Caspase-3/GSDME-dependent cell pyroptosis is elucidated. The implementation of the project will help the treatment window advance and promote the early prevention and treatment of mesangial proliferative glomerulonephritis.

系膜增生性肾小球肾炎(MsPGN)主要与免疫炎症反应相关，持续炎症及系膜增殖可导致终末期肾病发生，给社会和家庭造成巨大负担。故调控免疫炎症反应、抑制系膜增殖是其重要治疗策略。焦亡是由gasdermin介导的细胞程序性炎性坏死，可导致细胞内容物释放，激活强烈炎症反应。最新研究表明GSDME可被Caspase-3活化，诱导细胞焦亡。中药槐耳作为肿瘤辅助治疗药，有较好免疫调节及抑瘤作用，并可抑制大鼠系膜细胞异常增殖。本研究通过建立稳定模拟MsPGN的Thy-1肾炎大鼠模型，进行槐耳干预研究，利用MeDIP-seq及生物信息技术获取疾病进程中与焦亡及免疫炎症相关的甲基化差异基因，运用RNAi、质粒转染等技术开展体外细胞研究，以Caspase-3/GSDME依赖的细胞焦亡途径的免疫炎症调节机制为核心，阐述槐耳对MsPGN作用机制。项目的实施将有助于治疗窗前移，推进系膜增生性肾小球肾炎的早期防治工作。

慢性肾脏病患者人数日渐增多，其中最常见的肾脏疾病，系膜增生性肾小球肾炎(MsPGN)主要与免疫炎症反应相关，持续炎症及系膜增殖可导致终末期肾病发生，给社会和家庭造成巨大负担，且该类疾病缺乏理想治疗方案，故调控免疫炎症反应、抑制系膜增殖是其重要治疗策略。本项目通过建立了稳定模拟系膜增生性肾小球肾炎（MsPGN）的Thy-1肾炎大鼠模型，并进行槐耳干预研究，建立了急性系膜增生性肾小球肾炎大鼠研究平台。获取了一批不同时间点治疗组和模型差异蛋白。利用生物信息技术成功观察了目标基因GSDME在疾病及药物干预进程中的表达。通过体外细胞研究平台，验证了GSDME在系膜细胞焦亡、免疫炎症发生等过程中的作用。证实了中药槐耳通过影响肾脏GSDME甲基化水平，减少GSDME表达，激活Caspase-3/GSDME焦亡相关通路，改善免疫炎症细胞浸润，抑制系膜细胞增殖，从而有效防治MsPGN的发生发展。相关研究成果发表在国际期刊，并于2021年参与执笔起草的慢性肾脏病非透析期中西医结合诊疗专家共识中首次纳入了以槐耳为君药的中成药（槐杞黄颗粒）作为常用推荐用药之一。本项目的实施将有助于相关疾病的治疗窗前移，推进系膜增生性肾小球肾炎的早期防治工作。