枸杞多糖通过调控NLRP3/Caspase-1通路在抑制高糖诱导的HRMEC损伤中的作用  

Inhibiting effect of Lycium barbarum polysaccharide on high glucose-induced human retinal microvascular endothelial cell injury by regulating the NOD-like receptor family pyrin domain containing protein 3/Caspase-1 pathway

作　　者：张乐颖 王苏涵 秦婷婷 侯慧敏 王娇娇[1] 宋宗明 ZHANG Leying;WANG Suhan;QIN Tingting;HOU Huimin;WANG Jiaojiao;SONG Zongming(Department of Ophthalmology,Henan University People’s Hospital,Henan Provincial People’s Hospital,Henan Eye Hospital,Zhengzhou 450003,Henan Province,China;Xinxiang Medical University,Xinxiang 453003,Henan Province,China;Zhengzhou University People’s Hospital,Zhengzhou 450003,Henan Province,China)

机构地区：[1]河南大学人民医院,河南省人民医院,河南省立眼科医院眼科,河南省郑州市450003 [2]新乡医学院,河南省新乡市453003 [3]郑州大学人民医院,河南省郑州市450003

出　　处：《眼科新进展》2024年第11期857-862,共6页Recent Advances in Ophthalmology

基　　金：河南省科技攻关联合共建项目(编号:LHGJ20190817);中原科技领军人才项目(编号:224200510013);河南省重大科技专项(编号:221100310200)。

摘　　要：目的观察枸杞多糖(LBP)是否可以通过调控核苷酸寡聚化结构域样受体家族3/半胱天冬酶-1(NLRP3/Caspase-1)细胞焦亡途径抑制高糖诱导的人视网膜微血管内皮细胞(HRMEC)损伤。方法将体外培养的HRMEC随机分组,即正常组(给予5.5 mmol·L^(-1)葡萄糖)、高糖组(给予55.5 mmol·L^(-1)葡萄糖)、LBP低浓度组(给予55.5 mmol·L^(-1)葡萄糖+100 mg·L^(-1) LBP)、LBP中浓度组(给予55.5mmol·L^(-1)葡萄糖+500 mg·L^(-1) LBP)、LBP高浓度组(给予55.5 mmol·L^(-1)葡萄糖+1000 mg·L^(-1) LBP)、si-NC组(转染20μmol·L^(-1) si-NC后给予55.5 mmol·L^(-1)葡萄糖)和si-NLRP3组(转染20μmol·L^(-1) si-NLRP3后给予55.5 mmol·L^(-1)葡萄糖);利用CCK-8法检测各组HRMEC细胞增殖情况,流式细胞术检测各组HRMEC细胞焦亡情况,RT-PCR法检测各组HRMEC中NLRP3、Caspase-1、核因子(NF)-κB、Gasdermin-D(GSDMD)、血管内皮生长因子(VEGF)的mRNA相对表达水平,Western blot法检测各组HRMEC焦亡相关NLRP3、Caspase-1、NF-κB、GSDMD、VEGF的蛋白相对表达水平,ELISA检测各组HRMEC细胞上清液中细胞焦亡下游白细胞介素(IL)-1β和IL-18的表达水平。结果与正常组相比,高糖组HRMEC细胞增殖率降低,细胞焦亡率升高,NLRP3、Caspase-1、NF-κB、GSDMD、VEGF的mRNA和蛋白相对表达水平均升高,IL-1β和IL-18表达水平均升高(均为P<0.05);与高糖组相比,si-NLRP3组中HRMEC细胞增殖率升高,细胞焦亡率降低,NLRP3、Caspase-1、NF-κB、GSDMD、VEGF的mRNA和蛋白相对表达水平均降低,IL-1β和IL-18表达水平均降低(均为P<0.05);与高糖组相比,si-NC组的细胞增殖率,细胞焦亡率,NLRP3、Caspase-1、NF-κB、GSDMD、VEGF蛋白和mRNA以及IL-1β、IL-18的表达水平差异均无统计学意义(均为P>0.05);与高糖组相比,LBP中、高浓度组中HRMEC细胞增殖率升高,细胞焦亡率降低,NLRP3、Caspase-1、NF-κB、GSDMD、VEGF的mRNA和蛋白相对表达水平均降低,IL-1β和IL-18表达水平均降低(均为P<0.05);与�Objective To investigate if Lycium barbarum polysaccharide(LBP)could inhibit the high glucose-induced human retinal microvascular endothelial cell(HRMEC)injury by regulating the NOD-like receptor family pyrin domain containing protein 3(NLRP3)/Caspase-1 pyroptosis pathway.Methods HRMECs cultured in vitro were randomly divided into the control group(5.5 mmol·L^(-1) glucose),the high glucose group(55.5 mmol·L^(-1) glucose),the low LBP group(55.5 mmol·L^(-1) glucose+100 mg·L^(-1) LBP),the medium LBP group(55.5 mmol·L^(-1) glucose+500 mg·L^(-1) LBP),the high LBP group(55.5 mmol·L^(-1) glucose+1000 mg·L^(-1) LBP),the si-NC group(55.5 mmol·L^(-1) glucose after transfection with 20μmol·L^(-1) si-NC)and the si-NLRP3 group(55.5 mmol·L^(-1) glucose after transfection with 20μmol·L^(-1) si-NLRP3).The Cell Counting Kit-8 was used to detect the proliferation of HRMECs in each group and flow cytometry was adopted to measure the pyroptosis of HRMECs in each group.The reverse transcription-polymerase chain reaction was used to detect the relative messenger ribonucleic acid(mRNA)expression levels of NLRP3,Caspase-1,nuclear factor(NF)-κB,Gasdermin-D(GSDMD)and vascular endothelial growth factor(VEGF)in the HRMECs of each group,Western blot was adopted to detect the relative protein expression levels of HRMEC pyroptosis-related NLRP3,Caspase-1,NF-κB,GSDMD and VEGF in each group,and enzyme-linked immunosorbent assay was used to detect the interleukin(IL)-1βand IL-18 expression levels in downstream pyroptosis in the HRMEC supernatant of each group.Results Compared with the control group,the proliferation rate of HRMECs decreased,the pyroptosis rate increased,the relative mRNA and protein expression levels of NLRP3,Caspase-1,NF-κB,GSDMD and VEGF increased,and the expressions of IL-1βand IL-18 increased in the high glucose group(all P<0.05).Compared with the high glucose group,the proliferation rate of HRMECs increased,the pyroptosis rate decreased,the relative mRNA and protein expression levels of NLRP3,Caspase-1,NF-

关 键 词：枸杞多糖 NLRP3炎症小体 人视网膜微血管内皮细胞 细胞焦亡 

分 类 号：R774.1[医药卫生—眼科]