PMID- 34133251 OWN - NLM STAT- MEDLINE DCOM- 20220301 LR - 20220301 IS - 1460-2202 (Electronic) IS - 0271-3683 (Linking) VI - 47 IP - 1 DP - 2022 Jan TI - Protective Effect of Raf-1 Kinase Inhibitory Protein on Diabetic Retinal Neurodegeneration through P38-MAPK Pathway. PG - 135-142 LID - 10.1080/02713683.2021.1944644 [doi] AB - PURPOSE: This study aimed to investigate the effect of Raf-1 kinase inhibitory protein (RKIP) on diabetic retinal neurodegeneration in streptozotocin-treated rat model and high glucose-treated rat Muller cells. METHODS: Control and streptozotocin-treated rats were intravitreally injected with saline, RKIP gene overexpression lentivirus (oeRKIP) or negative control lentivirus (RKIP-vector). Normal or high glucose-treated Muller cells were transfected with saline, RKIP gene overexpression lentivirus or negative control lentivirus. Western blotting and immunofluorescence assay were utilized to evaluate the function of RKIP on the expression of RKIP, p38 mitogen-activated protein kinase (p38-MAPK), glutamate/aspartate transporter (GLAST), glutamine synthetase (GS), glial fibrillar acidic protein (GFAP) and cysteine-aspartic acid protease-3 (caspase-3). A glutamate assay kit was adopted to detect glutamate level in retina samples. Apoptosis of Muller cells was determined by Annexin-V/PI staining and flow cytometry. RESULTS: High glucose-treated Muller cells exhibited promoted apoptosis, while RKIP overexpression in high glucose-treated Muller cells down-regulated the enhanced apoptosis. Compared with rats injected with saline, streptozotocin-treated hyperglycemic rats displayed enhancement in the immunoreactivities of p38-MAPK and GFAP as well as in the protein expression of p38-MAPK and caspase-3. Strikingly, intravitreal injection of RKIP gene overexpression lentivirus in the hyperglycemic rats reversed the augmented immunoreactivities and protein expression mentioned above. Meanwhile, RKIP overexpression in the hyperglycemic rats improved the immunoreactivities and protein expression of RKIP, GS and GLAST. Besides, RKIP down-regulated the increased level of retinal glutamate in the hyperglycemic rats. CONCLUSIONS: Intravitreal injection of RKIP gene overexpression lentivirus functioned in preventing diabetic retinal neurodegeneration in a rat model of diabetes presumably by inhibiting p38-MAPK pathway. FAU - Wu, Chuanling AU - Wu C AD - Teaching and Research Section of Basic Medicine, Jianhu College, Zhejiang Industry Polytechnic College, Shaoxing, Zhejiang, China. AD - Department of Anatomy, Histology and Embryology, Jinzhou Medical University, Jinzhou, Liaoning, China. FAU - Xu, Kai AU - Xu K AD - Department of Hepatopancreatobiliary Surgery, Qinghai University Affiliated Hospital, Xining, Qinghai, China. FAU - Liu, Wenqiang AU - Liu W AD - Department of Anatomy, Histology and Embryology, Liaoning Key Laboratory of Diabetic Cognitive and Perceptive Dysfunction, Jinzhou, Liaoning, China. FAU - Liu, Anqi AU - Liu A AD - Department of Anatomy, Histology and Embryology, Liaoning Key Laboratory of Diabetic Cognitive and Perceptive Dysfunction, Jinzhou, Liaoning, China. FAU - Liang, Huimin AU - Liang H AD - Department of Fundus Disease, Shandong Lunan Eye Hospital, Linyi, Shangdong, China. FAU - Li, Qunwang AU - Li Q AD - Department of Anatomy, Histology and Embryology, Liaoning Key Laboratory of Diabetic Cognitive and Perceptive Dysfunction, Jinzhou, Liaoning, China. FAU - Feng, Zhen AU - Feng Z AD - Department of Anatomy, Histology and Embryology, Liaoning Key Laboratory of Diabetic Cognitive and Perceptive Dysfunction, Jinzhou, Liaoning, China. FAU - Yang, Yang AU - Yang Y AD - Department of Anatomy, Histology and Embryology, Liaoning Key Laboratory of Diabetic Cognitive and Perceptive Dysfunction, Jinzhou, Liaoning, China. FAU - Ding, Jiayuan AU - Ding J AD - Department of Anatomy, Histology and Embryology, Liaoning Key Laboratory of Diabetic Cognitive and Perceptive Dysfunction, Jinzhou, Liaoning, China. FAU - Zhang, Tianyi AU - Zhang T AD - Department of Anatomy, Histology and Embryology, Liaoning Key Laboratory of Diabetic Cognitive and Perceptive Dysfunction, Jinzhou, Liaoning, China. FAU - Liu, Yingxue AU - Liu Y AD - Department of Anatomy, Histology and Embryology, Liaoning Key Laboratory of Diabetic Cognitive and Perceptive Dysfunction, Jinzhou, Liaoning, China. FAU - Liu, Xuezheng AU - Liu X AD - Department of Anatomy, Histology and Embryology, Liaoning Key Laboratory of Diabetic Cognitive and Perceptive Dysfunction, Jinzhou, Liaoning, China. AD - Department of Anatomy, Histology and Embryology, Jinzhou Medical University, Jinzhou, Liaoning, China. FAU - Zuo, Zhongfu AU - Zuo Z AD - Department of Anatomy, Histology and Embryology, Jinzhou Medical University, Jinzhou, Liaoning, China. AD - Department of Anatomy, Histology and Embryology, Postdoctoral Research Station, Guangxi Medical University, Nanning, Guangxi, China. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20210726 PL - England TA - Curr Eye Res JT - Current eye research JID - 8104312 RN - EC 2.7.11.1 (Proto-Oncogene Proteins c-raf) SB - IM MH - Animals MH - *Apoptosis MH - *Diabetes Mellitus, Experimental MH - Diabetic Retinopathy/diagnosis/*genetics/metabolism MH - Disease Models, Animal MH - Ependymoglial Cells/*pathology MH - *Gene Expression Regulation MH - MAP Kinase Signaling System/*genetics MH - Proto-Oncogene Proteins c-raf/biosynthesis/*genetics MH - Rats MH - Rats, Sprague-Dawley OTO - NOTNLM OT - Muller cells OT - Raf-1 kinase inhibitory protein OT - diabetic retinal neurodegeneration OT - diabetic retinopathy OT - intravitreal injection OT - p38 mitogen-activated protein kinase EDAT- 2021/06/17 06:00 MHDA- 2022/03/03 06:00 CRDT- 2021/06/16 17:20 PHST- 2021/06/17 06:00 [pubmed] PHST- 2022/03/03 06:00 [medline] PHST- 2021/06/16 17:20 [entrez] AID - 10.1080/02713683.2021.1944644 [doi] PST - ppublish SO - Curr Eye Res. 2022 Jan;47(1):135-142. doi: 10.1080/02713683.2021.1944644. Epub 2021 Jul 26.