米诺环素调控MCPIP1/NLRP3通路抑制糖尿病视网膜病变细胞凋亡

Minocycline Regulates MCPIP1/NLRP3 Pathway and Inhibits Apoptosis in Diabetic Retinopathy

关键词:  米诺环素  糖尿病视网膜病变  凋亡  MCPIP1  NLRP3

Key words:  minocycline  diabetic retinopathy  apoptosis  MCPIP1  NLRP3

摘要:

目的 探讨米诺环素调控MCPIP1/NLRP3通路抑制糖尿病视网膜病变细胞凋亡的作用及其机制。方法 通过一次性腹腔注射链脲佐菌素的方法制备糖尿病小鼠模型，将模型小鼠分为模型组和米诺环素组，另设空白对照组。首先利用视网膜电图检测小鼠视网膜电位情况，再利用Western blotting检测小鼠视网膜中MCPIP1、NLRP3、ASC的表达水平，另利用TUNEL法对视网膜细胞的凋亡数目进行统计，以此检测米诺环素对糖尿病小鼠视网膜细胞的保护作用；而后以高糖培养的人视网膜上皮细胞为实验工具，利用RT-PCR检测用米诺环素处理后再敲低表达MCPIP1后NLRP3，ASC，IL-1β，IL-18，caspase-1的表达变化。结果 与模型组比较，米诺环素处理后，糖尿病小鼠视网膜电位振幅提升；MCPIP1的表达上调，NLRP3及ASC的表达受抑制；视网膜细胞凋亡数下降。米诺环素可以促进在高糖培养的人视网膜上皮细胞中MCPIP1的表达，而敲低表达MCPIP1可逆转米诺环素对NLRP3、ASC、IL-1β、caspase-1、IL-18的抑制作用。结论 米诺环素可通过上调MCPIP1以抑制NLRP3，从而对糖尿病视网膜病变起到抑制或延缓作用。

Abstract:

OBJECTIVE To investigate the effect of minocycline on MCPIP1/NLRP3 pathway in inhibiting apoptosis of diabetic retinopathy cells and its mechanism. METHODS Diabetic retinopathy(DR) mice models were prepared by intraperitoneal injection of streptozotocin, and then divided into model group and minocycline group, and set blank control group with no treatment. The retinal potential of mice was detected by electroretinogram(ERG). The expression levels of MCPIP1, NLRP3 and ASC in the retina of mice were detected by Western blotting. The number of apoptotic retinal cells was counted by TUNEL. Then the expression of NLRP3, ASC, IL-1β, IL-18 and caspase-1 treatment with minocycline and knockdown of MCPIP1 was detected by RT-PCR using high-sugar cultured human retinal pigment epithelium(RPE). RESULTS Compared with model group, following minocycline treatment, the amplitude of retinal potential in DR mice increased; the expression of MCPIP1 in the retina of DR mice was up-regulated, while the expression of NLRP3 and ASC was inhibited; the number of apoptotic retinal cells in DR mice decreased. Minocycline could promote the expression of MCPIP1 in RPE cultured with high glucose. Down-regulation of MCPIP1 reversed the inhibitory effects of minocycline on NLRP3, ASC, IL-1beta, caspase-1, IL-18. CONCLUSION Minocycline can inhibit NLRP3 by up-regulating MCPIP1, thus inhibiting or delaying diabetic retinopathy.