金丝桃苷对高糖诱导视网膜周细胞损伤的保护作用

Protective Effect of Hyperoside on Retinal Pericytes Injury Induced by High Glucose

目的 探讨金丝桃苷对高糖诱导视网膜周细胞(RPCs)损伤的保护作用,并对其可能机制做出初步探索。方法 将RPCs正常培养传代后接种至细胞培养板,按不同处理分为5组:正常葡萄糖(NG)组(葡萄糖5 mmol/L)、高糖(HG)组(葡萄糖20 mmol/L)、甘露醇(MT)组(葡萄糖5 mmol/L、渗透压20 mmol/L)、高糖+低浓度金丝桃苷(HG+L-HY)组(葡萄糖20 mmol/L、金丝桃苷100μg/mL)、高糖+高浓度金丝桃苷(HG+H-HY)组(葡萄糖20 mmol/L、金丝桃苷400μg/mL)。培育相应时间后,CCK-8法检测各组RPCs活性,Annexin V/PI及TUNEL法检测RPCs凋亡率,Real-time PCR及Western Blot法检测RPCs中NOD样受体蛋白3(NLRP3)、半胱天冬氨酸蛋白酶1(caspase-1)、Gasdermin-D蛋白-N(GSDMD-N)、白细胞介素-1β(IL-1β)及白细胞介素-18(IL-18)mRNA及蛋白表达水平。结果 与NG组比较,HG组RPCs活性降低,凋亡率升高(P<0.01),NLRP3、caspase-1、GSDMD-N、IL-1β、IL-18 mRNA及蛋白表达升高(P<0.01)。与HG组比较,HG+L-HY及HG+H-HY组RPCs活性升高,凋亡率降低(P<0.01),NLRP3、caspase-1、GSDMD-N、IL-1β、IL-18 mRNA及蛋白表达降低(P<0.05,P<0.01)。与HG+L-HY组比较,HG+H-HY组RPCs活性增高,凋亡率降低(P<0.01),NLRP3、caspase-1、GSDMD-N、IL-1β、IL-18 mRNA及蛋白表达降低(P<0.01)。结论 金丝桃苷可减轻高糖诱导的RPCs损伤,且该作用随金丝桃苷浓度提高而增强,其机制可能与抑制RPCs焦亡有关。

Objective To investigate the protective effect of hyperoside on retinal pericytes(RPCs)injury induced by high glucose and to explore the possible mechanism.Methods RPCs were cultured and transferred to cell culture plates.According to different treatments,RPCs were divided into 5 groups,the normal glucose(NG)group(5 mmol/L glucose),the high glucose(HG) group(20 mmol/L glucose),the mannitol(MT) group(5 mmol/L glucose,20 mmol/L osmotic pressure),the high glucose+low-concentration hyperoside(HG+L-HY) group(20 mmol/L glucose,100 μg/m L hyperoside),the high glucose + high-concentration hyperoside(HG+H-HY) group(20 mmol/L glucose,400 μg/m L hyperoside).The activity and apoptosis rate of RPCs were detected by CCK-8,Annexin V/PI and TUNEL.The protein and mRNA expression levels of NOD-like receptor protein 3(NLRP3),cysteine aspastic acid-specific protease-1(caspase-1),Gasdermin-D protein-N(GSDMD-N),interleukin-1β(IL-1β) and interleukin-18(IL-18) in RPCs were detected after corresponding incubation by Real-time PCR and Western Blot.Results Compared with NG group,the activity of RPCs decreased(P<0.01),the apoptosis rate increased(P<0.01),and the mRNA and protein expression levels of NLRP3,caspase-1,GSDMD-N,IL-1β and IL-18increased in the HG group(P<0.01).Compared with HG group,the RPCs activities of both HG+L-HY and HG+HHY groups increased(P<0.01),and the apoptosis rates decreased(P<0.01).The mRNA and protein expression levels of NLRP3,caspase-1,GSDMD-N,IL-1β and IL-18 decreased(P<0.05,P<0.01).Compared with HG+LHY group,the RPCs activities of HG+H-HY group increased(P<0.01),and the apoptosis rates decreased(P<0.01).The mRNA and protein expression levels of NLRP3,caspase-1,GSDMD-N,IL-1β and IL-18 decreased(P<0.01).Conclusion Hyperoside can concentration-dependently reduce RPCs injury induced by high glucose,and the mechanism may be related to the inhibition of RPCs pyroptosis.