【摘要】 目的 探究黄精多糖（PSP）对高糖诱导的晶状体上皮细胞（LECs）内质网应激（ERS）及凋亡的影响。方法 将SRA01/04细胞分为Control组、HG组和HG+PSP组。通过CCK-8法检测细胞活力以筛选PSP最佳干预浓度。建立大鼠晶状体体外培养模型，观察PSP对晶状体混浊的影响。采用Western blot检测磷酸化蛋白激酶样内质网激酶（p-PERK）、蛋白激酶样内质网激酶（PERK）、转录因子C/EBP同源蛋白（CHOP）、葡萄糖调节蛋白78（GRP78）等ERS相关蛋白及B细胞淋巴瘤/白血病-2（Bcl-2）和Bcl-2相关X蛋白（Bax）等凋亡蛋白的表达。采用免疫荧光染色法检测GRP78的表达与分布，TUNEL法检测细胞凋亡。结果 CCK-8法检测结果显示，与Control组相比，HG组细胞活力降低（P<0.001）;与HG组相比，不同浓度PSP干预后细胞活力有不同程度的回升，其中PSP浓度为25.00μmol·L^-1时细胞活力升高最显著（P<0.01）。晶状体体外培养结果显示，HG组晶状体混浊面积占比较Control组明显升高（P<0.001）,而HG+PSP组较HG组显著降低（P<0.01）。Western blot检测结果显示，与Control组相比，HG组p-PERK/PERK、GRP78及CHOP蛋白表达均升高，Bcl-2/Bax比值降低（均为P<0.05）;与HG组相比，HG+PSP组上述ERS相关蛋白表达均显著降低，Bcl-2/Bax比值显著升高（均为P<0.05）。免疫荧光染色结果显示，与Control组相比，HG组GRP78荧光强度显著增加（P<0.01）;与HG组相比，HG+PSP组荧光强度显著降低（P<0.01）。TUNEL检测结果表明，HG组细胞凋亡水平较Control组升高，加入PSP后凋亡降低（均为P<0.05）。结论 PSP能够通过抑制高糖诱导LECs的ERS和细胞凋亡，从而延缓晶状体混浊的发生。更多还原

【Abstract】 Objective To investigate the effects of Polygonatum sibiricum polysaccharide（PSP） on endoplasmic reticulum stress（ERS） and apoptosis of lens epithelial cells（LECs） induced by high glucose（HG）.Methods SRA01/04 cells were divided into the Control group, HG group, and HG+PSP group.Cell viability was measured by the Cell Counting Kit-8（CCK-8） assay to determine the optimal concentration of PSP.An in vitro rat lens culture model was established to observe the effect of PSP on lens opacity.The expression levels of ERS-related proteins, including phosphorylated protein kinase-like endoplasmic reticulum kinase（p-PERK）,protein kinase-like endoplasmic reticulum kinase（PERK）,transcription factor C/EBP homologous protein（CHOP）,and glucose-regulated protein 78（GRP78）,as well as apoptosis-related proteins, including B-cell lymphoma/leukemia-2（Bcl-2） and Bcl-2-associated X protein（Bax） were detected by Western blot.The expression and distribution of GRP78 were measured by immunofluorescence staining, and cell apoptosis was quantified by TUNEL assay.Results CCK-8 assay showed that compared with the Control group, cell viability in the HG group significantly decreased（P<0.001）.Compared with the HG group, cell viability increased to varying degrees after intervention with different concentrations of PSP,with the most significant increase observed at a PSP concentration of 25.00 μmol·L^-1（P<0.01）.In the in vitro lens culture, the percentage of lens opacity in the HG group was significantly higher than that in the Control group（P<0.001）,while it decreased significantly in the HG+PSP group compared with the HG group（P<0.01）.Western blot results revealed that compared with the Control group, the expression levels of p-PERK/PERK,GRP78,and CHOP proteins were up-regulated, while the Bcl-2/Bax ratio was down-regulated in the HG group（all P<0.05）.Compared with the HG group, the expression levels of the aforementioned ERS-related proteins significantly decreased, while the Bcl-2/Bax ratio significantly increased in the HG+PSP group（all P<0.05）.Immunofluorescence staining showed that the fluorescence intensity of GRP78 in the HG group was significantly higher than that in the Control group（P<0.01）,while it was significantly reduced in the HG+PSP group compared with the HG group（P<0.01）.TUNEL assay demonstrated that the apoptosis level in the HG group increased compared with the Control group, which was effectively attenuated by PSP treatment（all P<0.05）.Conclusion PSP can delay the progression of lens opacity by inhibiting HG-induced ERS and apoptosis in LECs.更多还原