舒洛地特对高糖诱导的人视网膜微血管内皮细胞功能紊乱的作用及其机制

Effects of Sulodexide on High Glucose-induced Dysfunctionin Human Retinal Endothelial Cells

【目的】通过观察体外高糖环境下培养的原代人视网膜微血管内皮细胞(HREC)中C1q/TNF相关蛋白3(CTRP3)及炎症因子的表达情况及舒洛地特对其表达的影响,探讨舒洛地特治疗糖尿病视网膜病变的机制。【方法】将体外培养的HREC分为以下4组:对照组(普通内皮细胞培养基,含5.5 mmol/L葡萄糖),甘露醇组(24.5 mmol/L甘露醇+5.5mmol/L葡萄糖),高糖组(30 mmol/L葡萄糖),舒洛地特干预组(0.25、0.5、1.0 LRU/m L舒洛地特+30 mmol/L葡萄糖)。采用Western blot法检测各组CTPR3、TNF-α、MCP-1及p-NF-κB p65蛋白的表达。【结果】高糖培养HREC,见CTRP3蛋白表达水平随高糖培养时间的延长而逐步增加并在干预8 h时达到峰值(P<0.01),当高糖培养并加入舒洛地特干预后CTRP3的蛋白表达水平与高糖组相比呈剂量依赖性下降(P<0.05)。此外,高糖培养12h后TNF-α、MCP-1及p-NF-κB p65蛋白表达水平较对照组明显升高(P<0.05),而当高糖培养下加入舒洛地特干预后,TNF-α、MCP-1及p-NF-κB p65蛋白表达水平显著低于高糖组(P<0.05)。对照组与甘露醇组间CTPR3、TNF-α、MCP-1及p-NF-κB p65蛋白表达水平差异均无统计学意义(P>0.05)。【结论】本研究首次证实了高糖可使HREC中CTRP3蛋白表达上调,舒洛地特可抑制高糖引起的CTRP3蛋白表达增加;舒洛地特可抑制高糖诱导的HREC中转录因子p-NF-κB p65及炎症因子TNF-α、MCP-1的表达水平升高,这可能是其治疗糖尿病视网膜病变的机制之一。

[Objective] To investigate the effects of sulodexide on the expression of CTRP3 and inflammatory cytokines in high glucose cultured HREC. [Methods] The HREC were divided into 4 groups as follow： control group （treated with 5.5 mmol/L glucose）, mannitol group （treated with 24.5 mmol/L mannitol ＋5.5 mmol/L glucose）, high glucose group （treated with 30 mmol/L glucose） and sulodexide group （0.25, 0.5, 1.0 LRU/mL sulodexide ＋ 30 mmol/L glucose）. The expression levels of CTRP3, MCP-1 , TNF-α and p-NF-KB p65 were determined by Western blot analysis. [ Results] After treatment with high glucose, the protein level of CTRP3 was increased in a time-dependent manner in HREC, and was peaked at 8 hours （P 〈 0.01）, but sulodexide could significantly reduce this hyperglucose-related response in a dose-dependent manner （P 〈 0.05）. Additional, the protein level of MCP-1, TNF-α and p-NF-κB p65were significantly increased after high glucose challenge （P 〈 0.05）, and significantly decreased after exposure to sulodexide （P 〈 0.05）. However, no significant differences of CTRP3, MCP-1, TNF-α and p-NF-κB p65 were found between control group and mannitol group （P 〉 0.05）. [Conclusion] High glucose could up-regulate the expression of CTRP3 and the inflammatory response in HREC, while sulodexide could inhibit these changes, which may provide a potential mechanism of sulodexide in treating diabetic retinopathy.