白术多糖通过novel-mir2双靶向TRADD/MLKL缓解免疫抑制雏鹅的胸腺程序性坏死

程序性坏死导致胸腺的发育及功能被抑制，前期研究发现，白术多糖可缓解雏鹅胸腺程序性坏死，且novel-mir2表达量与程序性坏死水平负相关，然而novel-mir2在白术多糖缓解免疫抑制雏鹅胸腺程序性坏死中的机制尚不清楚。预测novel-mir2双靶向调控TRADD和MLKL，因此本研究应用双荧光素酶报告基因法验证其靶向关系；构建免疫抑制雏鹅胸腺程序性坏死模型并添加白术多糖，结合淋巴细胞程序性坏死诱导试验、TRADD和/或MLKL双过表达试验，应用电镜、免疫荧光、Elisa、qPCR、免疫印迹、流式细胞术等，通过形态学观察、程序性坏死水平、抗氧化指标、细胞因子、novel-mir2、靶基因及程序性坏死通路基因的表达量，阐明白术多糖通过novel-mir2缓解免疫抑制雏鹅胸腺程序性坏死的机制，为开发植物多糖作为添加剂提供理论依据，为比较医学提供借鉴。

Necroptosis causes the development and function of the thymus to be inhibited. Preliminary studies have found that PAMK can relieve necroptosis in gosling thymus, and the expression of novel-mir2 is negatively correlated with the level of necroptosis. However, the mechanism of novel-mir2 in PAMK relieving immunosuppressive gosling thymus necroptosis still not clear. It is predicted that novel-mir2 dual-targeted regulation of TRADD and MLKL. So this study applied dual luciferase reporter gene method to verify its targeting relationship; constructed an immunosuppressive gosling thymus necroptosis model and added PAMK, combined with lymphocyte necroptosis induction test, TRADD and / or MLKL double overexpression test to analysis morphological observation, necroptosis level, antioxidant indicators, cytokines, novel-mir2, the expression of target genes and necroptosis pathway genes by electron microscopy, immunofluorescence, Elisa, qPCR, western blot, flow cytometry, etc. The purpose of the study was to clarify the mechanism of PAMK through novel-mir2 to relieve the immunosuppressive gosling thymus necroptosis, provide a theoretical basis for the development of plant polysaccharides as additives, and provide reference for comparative medicine.