基于SIRT6/AKT/mTOR自噬通路研究枸杞多糖对糖尿病性白内障的保护机制

白内障是糖尿病的严重并发症之一，其发病机制尚未完全阐明，既往研究证实SIRT6可通过对自噬相关蛋白的去乙酰化修饰调节细胞自噬水平，但在糖尿病性白内障中SIRT6是否能够调节晶状体上皮细胞的自噬水平尚未见报道。项目组前期研究发现糖尿病大鼠晶状体组织SIRT6的表达下降，同时体外高糖培养的晶状体上皮细胞活性下降，而枸杞多糖能够促进SIRT6的表达并增加细胞活性。本研究将建立糖尿病白内障大鼠模型，观察SIRT6及其下游AKT、mTOR等细胞因子的表达情况；同时利用基因重组技术构建SIRT6过表达和沉默两种载体，观察外源性SIRT6表达增强和抑制后对糖尿病大鼠晶状体及高糖培养的晶状体上皮细胞的影响，并检测SIRT6/AKT/mTOR通路中细胞因子的表达水平，进而在整体和细胞水平研究枸杞多糖干预后SIRT6/AKT/mTOR通路相关细胞因子的表达，从而阐明枸杞多糖改善糖尿病性白内障的药效及分子机制。

Cataract is one of the serious complications of diabetes mellitus, and its pathogenesis has not been fully elucidated. Previous studies have confirmed that SIRT6 can regulate the autophagy level by deacetylation of autophagy related proteins, but whether SIRT6 can regulate the autophagy level of lens epithelial cells in diabetic cataract has not been reported. In the prophase study of the project group, it was found that the expression of SIRT6 in the lens of diabetic rats decreased, while the activity of lens epithelial cells cultured in high glucose in vitro decreased, while Lycium barbarum polysaccharides could promote the expression of SIRT6 and increase the activity of cells. In this study, we will establish a diabetic cataract rat model, observe the expression of SIRT6 and its downstream AKT, mTOR and other cytokines. At the same time, we will construct two kinds of SIRT6 overexpression and silencing vectors by gene recombination technology, observe the effect of enhanced and inhibited expression of exogenous SIRT6 on the lens in diabetic rats and lens epithelial cells cultured with high glucose , and detect the cytokines in SIRT6/AKT/mTOR pathway. The expression of cytokines in SIRT6/AKT/mTOR pathway after Lycium barbarum polysaccharides intervention will be studied at the whole and cell level, so as to clarify the efficacy and molecular mechanism of Lycium barbarum polysaccharides in improving diabetic cataract and provide experimental basis and theoretical basis for its clinical application.