新型香菇菌丝体多糖通过lncRNA-MALAT1/NF-κB/NLRP3通路抑制晚期糖基化终末产物（AGEs）介导血管内皮细胞焦亡作用研究

糖尿病血管病变(DA)是常见的糖尿病并发症之一，晚期糖基化终末产物(AGEs)在其发生发展中起着重要作用。寻找有效抑制AGEs生成和细胞毒性的天然产物，对于防治DA具有重要意义。食药用真菌是天然产物资源的重要组成部分。实验室前期制备的一种新型香菇菌丝体多糖（LMP）具有较好抗氧化作用。本项目以人脐静脉内皮细胞为研究对象，模拟糖尿病病理条件，检测lncRNA-MALAT1表达、活性氧水平、焦亡率等指标，考察LMP抑制AGEs诱导细胞焦亡作用；双荧光素酶报告基因、RIP、qRT-PCR、Western blot等方法研究lncRNA-MALAT1/NF-κB/NLRP3通路作用；最后在动物水平上评价LMP对AGEs毒性的抑制效果。采用分子生物学、细胞生物学、分子模拟等技术，从分子、细胞、动物水平系统阐明LMP抑制AGEs诱导血管内皮细胞焦亡作用及机制，为真菌资源开发和DA防治提供重要实验依据。

Diabetes angiopathy (DA) is one of the common diabetic complications. Advanced glycation end products (AGEs) play an important role in the development of DA. Finding natural products that can effectively inhibit the generation of AGEs and cytotoxicity is of great significance for the DA prevention. Edible and medicinal fungi is an important component of natural product resources. A novel polysaccharide was obtained from Lentinus edodes mycelium (LMP) in our previous studies. It has good antioxidant effect. HUVECs will be taken as the object and pathological condition of diabetes mellitus will be simulated with AGEs. The expression of lncRNA-MALAT1, cellular reactive oxygen species, cell pyroptosis rate and other indexes will be detected and the inhibition of LMP on AGEs-induced damage in HUVECs will be investigated. The function of lncRNA-MALAT1/NF-κB/NLRP3 signal path in LMP inhibiting AGEs-induced pyroptosis will be further studied by dual-luciferase reporter gene, RIP, qRT-PCR, immunofluorescence, western blot and so on. At last, the inhibition of LMP on the toxicity of AGEs will be evaluated in animal model. Using of molecular biology, cell biology, molecular simulation technologies, researches will give a systematically illustration of the LMP inhibiting pyroptosis of vascular endothelial cells induced by AGEs on the molecular, cellular and animal levels. This study will provide the scientific basis for the fungus resource development and important experimental evidence of DA prevention.