[关键词]
[摘要]
目的:探讨脂联素(ADPN)对高糖(HG)环境下人视网膜微血管内皮细胞(hRMECs)血管生成的影响,以及 NOD 样受体家族含 pyrin 结构域蛋白 3(NLRP3)炎症小体的作用。
方法:将人视网膜微血管内皮细胞分为6组:对照组(不做处理); 高糖组:用 D-葡萄糖孵育; 脂联素组:先用脂联素预处理,再用 D-葡萄糖孵育; CY-09组:先用NLRP3抑制剂CY-09预处理,再用 D-葡萄糖孵育; 尼日利亚菌素组:先用NLRP3激活剂尼日利亚菌素预处理,再用 D-葡萄糖孵育; 尼日利亚菌素+脂联素组:先后用尼日利亚菌素和脂联素预处理,再用 D-葡萄糖孵育。采用蛋白质印迹法(Western blot)检测NLRP3水平; 细胞划痕实验检测人视网膜微血管内皮细胞迁移能力; 基质胶(Matrigel)实验检测细胞管样形成能力。
结果:高糖环境下人视网膜微血管内皮细胞中NLRP3表达显著升高(P<0.01),而脂联素和 CY-09 可降低升高的NLRP3(与高糖组比较,均P<0.05)。尼日利亚菌素可显著升高NLRP3水平(与对照组比较,P<0.01),而脂联素可逆转该作用(与尼日利亚菌素组比较,P=0.032)。高糖组细胞迁移能力增强(P<0.001),总主干段长度和网格数增加(P<0.001); 脂联素组和CY-09组上述指标均降低(与高糖组比较,均P<0.01)。尼日利亚菌素可显著提高高糖环境下细胞迁移及管样形成能力(总主干段长度、网格数)(P=0.003),而脂联素可逆转该变化。
结论:脂联素可改善高糖条件下人视网膜微血管内皮细胞的迁移及血管生成。
[Key word]
[Abstract]
AIM: To explore the effect of adiponectin(ADPN)on angiogenesis of human retinal microvascular endothelial cells(hRMECs)in high glucose(HG)environment and role of NOD-like receptor family pyrin domain containing 3(NLRP3)inflammasome.
METHODS: The hRMECs were divided into six groups, including control group(without treatment), HG group: incubated with D-glucose, ADPN group: pretreatment with ADPN and then incubated with D-glucose, CY-09 group: pretreatment with CY-09(an NLRP3 inhibitor)and then incubated with D-glucose, Nigericin group: pretreatment with nigericin(an NLRP3 activator)and then incubated with D-glucose, Nigericin+ADPN group: pretreatment with nigericin and ADPN and then incubated with D-glucose. NLRP3 level was detected using Western blot analysis. hRMECs migration was measured using scratch wound healing assay. The tube formation of hRMECs was detected using Matrigel.
RESULTS: The NLRP3 expression in hRMECs cultured in an HG environment was significantly increased(P<0.01), while ADPN and CY-09 reduced the elevated NLRP3(both P<0.05 vs HG group). Nigericin significantly increased NLRP3 levels(P<0.01 vs control group)which was reversed by ADPN(P=0.032 vs Nigericin group). hRMECs migration ability(P<0.001), and total master segments length and number of meshes increased in HG group(P<0.001)while decreased in ADPN and CY-09 groups(all P<0.01 vs HG group). The hRMECs migration ability and tube formation(total master segments length and number of meshes)in HG environment were significantly increased by nigericin(P=0.003), while ADPN inversed the change.
CONCLUSION: ADPN alleviates the migration and angiogenesis of hRMECs under HG conditions.
[中图分类号]
[基金项目]
陕西省科学技术厅自然科学基础研究计划项目(No.2021JM-490)