Is Iba-1 protein expression a sensitive marker for microglia activation in experimental diabetic retinopathy?
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Guo-Xu Xu. Department of Ophthalmology, the Second Affiliated Hospital of Soochow University, 1055 Sanxiang Road, Outpatient Building, 4th Floor, Room E10, Suzhou 215004, Jiangsu Province, China. phacoxu@126.com; Jing-Fa Zhang. Department of Ophthalmology, Shanghai General Hospital (Shanghai First People’s Hospital), Shanghai Jiao Tong University, 100 Haining Road, Hongkou District, Shanghai 200080, China. 13917311571@139.com

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Supported by National Natural Science Foundation of China (No.81570852).

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    Abstract:

    AIM: To investigate the changes of Iba-1 and other potential markers for microglia activation in experimental diabetic retinopathy (DR). METHODS: Male Sprague-Dawley rats were rendered diabetes via intraperitoneal injection of streptozotocin. The retinas were harvested at 1 to 24wk after diabetes onset. Hypoxia-treated mouse microglial cell line (BV2 cells) was employed as the in vitro model to mimic diabetic condition. The expressions of Iba-1, CD11b, ICAM-1 as well as the inflammatory factors were examined with real-time polymerase chain reaction, Western blot and immunofluorescence both in vivo and in vitro. RESULTS: Compared with age-matched normal control, the number of microglia (Iba-1 positive immunostaining) in diabetic rat retinas was increased from 1 to 24wk of diabetes, which was most obvious at 12wk of diabetes. Iba-1 protein expression detected by Western blot was increased slightly in diabetic rat retinas compared with that in age-matched normal control; however, there was statistically significant between two groups only at 2wk after diabetes onset. The mRNA expression of Iba-1 was decreased significantly at 2 and 4wk of diabetic rat retinas, and remained unchanged at 8 and 12wk of diabetes. In BV2 cells, there was no significant change for the Iba-1 protein expression between normoxia and hypoxia groups; however, its mRNA level was decreased significantly under hypoxia. To further characterize microglial activation, F4/80, CD11b and inflammatory factors were detected both in vivo and in vitro. Compared with normal control, the expressions of F4/80 and CD11b as well as the inflammatory factors, such as ICAM-1, iNOS, COX2, IL-1β and IL-6, were increased significantly both in vivo and in vitro. CONCLUSION: Iba-1 protein expression might not be a sensitive marker to evaluate the activation of microglia in experimental DR. However, Iba-1 immunostaining, in combination with other markers like CD11b and ICAM-1, could be well reflect the activation of microglia. Thus, it is of great importance to explore other potential marker to evaluate the activation of microglia.

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Fan-Jun Shi, Hai Xie, Chao-Yang Zhang, et al. Is Iba-1 protein expression a sensitive marker for microglia activation in experimental diabetic retinopathy?. Int J Ophthalmol, 2021,14(2):200-208

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Publication History
  • Received:June 16,2020
  • Revised:October 10,2020
  • Adopted:
  • Online: December 24,2020
  • Published: