Yun Ou-Yang1, Zheng-Li Liu1, Chun-Long Xu1, Jia-Liang Wu1, Jun Peng2, Qing-Hua Peng3. 1. Yancheng Affiliated Hospital of Nanjing University of Chinese Medicine, Jiangsu, China. 2. Hunan University of Chinese Medicine, Hunan, China. Electronic address: 154451101@qq.com. 3. Hunan University of Chinese Medicine, Hunan, China. Electronic address: pqh410007@163.com.
Abstract
PURPOSE: Glaucoma is an eye disease characterized by the loss of peripheral vision, high pressure in the eye, optic nerve damage, and the loss of peripheral vision due to loss of retinal ganglion cells (RGCs). A number of miRNA have been detected in the pathogenesis of glaucoma. The paper was to focus on the miR-223 in RGCs apoptosis and inflammation, and investigated the possible mechanisms in vitro and in vivo. MATERIALS AND METHODS: After miR-223 inhibitor and mimics transfected into RGCs, the expression of miR-223 was detected by QRT-PCR, cell proliferation were performed by CCK-8 and EdU assays, cell apoptosis were measured by flow cytometer and TUNEL assays, apoptosis and inflammation -related proteins were detected by western blot, and whether miR-223 target to HSP-70 was detected by Luciferease reporter assay. Moreover, the effects si-HSP-70 on RGCs or RGCs transfected with miR-223 inhibitor were detected. In vivo study. New Zealand White rabbits (20 females) were used to detect the effect of miR-223 on the rabbit glaucoma model induced by injection of carbomer. RESULTS: CCK-8 and EdU assays demonstrated that miR-223 mimics decreased RGCs proliferation. FITC-Annexin V/PI Apoptosis and TUNEL assays showed that miR-223 mimics induced RGCs apoptosis. Western blot revealed that miR-223 mimics enhanced the expression of relative apoptosis and inflammation factors. Further, we demonstrated that miR-223 could inhibit cell proliferation, induce cell apoptosis as well as inflammation by targeting HSP-70 in RGCs. Moreover, the results were confirmed in rabbit glaucoma model. CONCLUSIONS: In summary, miR-223 plays a vital role in RGCs by regulating HSP-70 expression, and the new therapeutic strategy might potentially contribute to benefit glaucoma treatment.
PURPOSE:Glaucoma is an eye disease characterized by the loss of peripheral vision, high pressure in the eye, optic nerve damage, and the loss of peripheral vision due to loss of retinal ganglion cells (RGCs). A number of miRNA have been detected in the pathogenesis of glaucoma. The paper was to focus on the miR-223 in RGCs apoptosis and inflammation, and investigated the possible mechanisms in vitro and in vivo. MATERIALS AND METHODS: After miR-223 inhibitor and mimics transfected into RGCs, the expression of miR-223 was detected by QRT-PCR, cell proliferation were performed by CCK-8 and EdU assays, cell apoptosis were measured by flow cytometer and TUNEL assays, apoptosis and inflammation -related proteins were detected by western blot, and whether miR-223 target to HSP-70 was detected by Luciferease reporter assay. Moreover, the effects si-HSP-70 on RGCs or RGCs transfected with miR-223 inhibitor were detected. In vivo study. New Zealand White rabbits (20 females) were used to detect the effect of miR-223 on the rabbit glaucoma model induced by injection of carbomer. RESULTS: CCK-8 and EdU assays demonstrated that miR-223 mimics decreased RGCs proliferation. FITC-Annexin V/PI Apoptosis and TUNEL assays showed that miR-223 mimics induced RGCs apoptosis. Western blot revealed that miR-223 mimics enhanced the expression of relative apoptosis and inflammation factors. Further, we demonstrated that miR-223 could inhibit cell proliferation, induce cell apoptosis as well as inflammation by targeting HSP-70 in RGCs. Moreover, the results were confirmed in rabbit glaucoma model. CONCLUSIONS: In summary, miR-223 plays a vital role in RGCs by regulating HSP-70 expression, and the new therapeutic strategy might potentially contribute to benefit glaucoma treatment.
Authors: Nilisha Fernando; Josephine H C Wong; Shannon Das; Catherine Dietrich; Riemke Aggio-Bruce; Adrian V Cioanca; Yvette Wooff; Joshua A Chu-Tan; Ulrike Schumann; Chinh Ngo; Rohan W Essex; Camilla Dorian; Sarah A Robertson; Si Ming Man; Jan Provis; Riccardo Natoli Journal: Front Cell Dev Biol Date: 2020-06-26