BACKGROUND Diabetic retinopathy(DR)is a major ocular complication of diabetes mellitus,leading to visual impairment.Retinal pigment epithelium(RPE)injury is a key component of the outer blood retinal barrier,and its d...BACKGROUND Diabetic retinopathy(DR)is a major ocular complication of diabetes mellitus,leading to visual impairment.Retinal pigment epithelium(RPE)injury is a key component of the outer blood retinal barrier,and its damage is an important indicator of DR.Receptor for activated C kinase 1(RACK1)activates protein kinase C-ε(PKC-ε)to promote the generation of reactive oxygen species(ROS)in RPE cells,leading to apoptosis.Therefore,we hypothesize that the activation of RACK1 under hypoxic/high-glucose conditions may promote RPE cell apoptosis by modulating PKC-ε/ROS,thereby disrupting the barrier effect of the outer blood retinal barrier and contributing to the progression of DR.AIM To investigate the role and associated underlying mechanisms of RACK1 in the development of early DR.METHODS In this study,Sprague-Dawley rats and adult RPE cell line-19(ARPE-19)cells were used as in vivo and in vitro models,respectively,to explore the role of RACK1 in mediating PKC-εin early DR.Furthermore,the impact of RACK1 on apoptosis and barrier function of RPE cells was also investigated in the former model.RESULTS Streptozotocin-induced diabetic rats showed increased apoptosis and upregulated expression of RACK1 and PKC-εproteins in RPE cells following a prolonged modeling.Similarly,ARPE-19 cells exposed to high glucose and hypoxia displayed elevated mRNA and protein levels of RACK1 and PKC-ε,accompanied by an increases in ROS production,apoptosis rate,and monolayer permeability.However,silencing RACK1 significantly downregulated the expression of PKC-εand ROS,reduced cell apoptosis and permeability,and protected barrier function.CONCLUSION RACK1 plays a significant role in the development of early DR and might serve as a potential therapeutic target for DR by regulating RPE apoptosis and barrier function.展开更多
目的以GEO(Gene Expression Omnibus)数据库为基础分析活化的蛋白激酶C受体1(receptor for activated C kinase1,RACK1)在糖尿病肾小管病中的表达及可能的机制。方法通过搜索Nephroseq数据库及KEGG(Kyoto Encyclopedia of Genes and Gen...目的以GEO(Gene Expression Omnibus)数据库为基础分析活化的蛋白激酶C受体1(receptor for activated C kinase1,RACK1)在糖尿病肾小管病中的表达及可能的机制。方法通过搜索Nephroseq数据库及KEGG(Kyoto Encyclopedia of Genes and Genomes),明确RACK1在人的糖尿病肾小管病中表达情况及可能的通路。将培养的肾小管上皮细胞分为对照组、甘露醇组和高糖组,实时定量PCR测定其下游靶基因STAT1和STAT3的变化。FVB/N小鼠分为对照组和糖尿病肾病组,糖尿病肾病组小鼠给予腹腔注射链脲佐菌素构建糖尿病肾病的模型,实时定量PCR测定STAT1和STAT3 mRNA水平,免疫荧光染色明确p-STAT1和p-STAT3的表达情况。结果数据库数据分析发现RACK1在人糖尿病肾小管中表达明显升高。KEGG提示RACK1可能通过激活Jak-STAT信号通路发挥作用。与对照组比较,高糖组肾小管上皮细胞的RACK1、STAT1及STAT3的mRNA水平均明显升高,差异有统计学意义(P<0.05)。糖尿病肾病动物模型中,肾小管的RACK1、STAT1及STAT3的mRNA水平均明显升高,差异有统计学意义(P<0.05);免疫荧光示p-STAT1阳性,主要位于肾小管上皮细胞中。结论 RACK1在糖尿病肾小管中高表达,可能是通过磷酸化STAT1激活Jak-STAT信号通路参与糖尿病肾小管病的发生发展。展开更多
The receptor for activated C-kinase 1 （RACK1） is a highly conserved scaffold protein with versatile functions, and plays important roles in the regulation of plant growth and development. Transgenic rice plants, in ...The receptor for activated C-kinase 1 （RACK1） is a highly conserved scaffold protein with versatile functions, and plays important roles in the regulation of plant growth and development. Transgenic rice plants, in which the expression of RACK1 gene was inhibited by RNA interference （RNAi）, were studied to elucidate the possible functions of RACK1 in responses to drought stress in rice. Real-time PCR analysis showed that the expression of RACK1 in transgenic rice plants was inhibited by more than 50%. The tolerance to drought stress of the transgenic rice plants was higher as compared with the non-transgenic rice plants. The peroxidation of membrane and the production of malondialdehyde were significantly lower and the superoxide dismutase activity in transgenic rice plants was significantly higher than those in non-trangenic rice plants It is suggested that RACK1 negatively regulated the redox system-related tolerance to drought stress of rice plants.展开更多
基金Supported by National Natural Science Foundation of China,No.82260211Key Research and Development Project in Jiangxi Province,No.20203BBG73058Chinese Medicine Science and Technology Project in Jiangxi Province,No.2020A0166.
文摘BACKGROUND Diabetic retinopathy(DR)is a major ocular complication of diabetes mellitus,leading to visual impairment.Retinal pigment epithelium(RPE)injury is a key component of the outer blood retinal barrier,and its damage is an important indicator of DR.Receptor for activated C kinase 1(RACK1)activates protein kinase C-ε(PKC-ε)to promote the generation of reactive oxygen species(ROS)in RPE cells,leading to apoptosis.Therefore,we hypothesize that the activation of RACK1 under hypoxic/high-glucose conditions may promote RPE cell apoptosis by modulating PKC-ε/ROS,thereby disrupting the barrier effect of the outer blood retinal barrier and contributing to the progression of DR.AIM To investigate the role and associated underlying mechanisms of RACK1 in the development of early DR.METHODS In this study,Sprague-Dawley rats and adult RPE cell line-19(ARPE-19)cells were used as in vivo and in vitro models,respectively,to explore the role of RACK1 in mediating PKC-εin early DR.Furthermore,the impact of RACK1 on apoptosis and barrier function of RPE cells was also investigated in the former model.RESULTS Streptozotocin-induced diabetic rats showed increased apoptosis and upregulated expression of RACK1 and PKC-εproteins in RPE cells following a prolonged modeling.Similarly,ARPE-19 cells exposed to high glucose and hypoxia displayed elevated mRNA and protein levels of RACK1 and PKC-ε,accompanied by an increases in ROS production,apoptosis rate,and monolayer permeability.However,silencing RACK1 significantly downregulated the expression of PKC-εand ROS,reduced cell apoptosis and permeability,and protected barrier function.CONCLUSION RACK1 plays a significant role in the development of early DR and might serve as a potential therapeutic target for DR by regulating RPE apoptosis and barrier function.
文摘目的以GEO(Gene Expression Omnibus)数据库为基础分析活化的蛋白激酶C受体1(receptor for activated C kinase1,RACK1)在糖尿病肾小管病中的表达及可能的机制。方法通过搜索Nephroseq数据库及KEGG(Kyoto Encyclopedia of Genes and Genomes),明确RACK1在人的糖尿病肾小管病中表达情况及可能的通路。将培养的肾小管上皮细胞分为对照组、甘露醇组和高糖组,实时定量PCR测定其下游靶基因STAT1和STAT3的变化。FVB/N小鼠分为对照组和糖尿病肾病组,糖尿病肾病组小鼠给予腹腔注射链脲佐菌素构建糖尿病肾病的模型,实时定量PCR测定STAT1和STAT3 mRNA水平,免疫荧光染色明确p-STAT1和p-STAT3的表达情况。结果数据库数据分析发现RACK1在人糖尿病肾小管中表达明显升高。KEGG提示RACK1可能通过激活Jak-STAT信号通路发挥作用。与对照组比较,高糖组肾小管上皮细胞的RACK1、STAT1及STAT3的mRNA水平均明显升高,差异有统计学意义(P<0.05)。糖尿病肾病动物模型中,肾小管的RACK1、STAT1及STAT3的mRNA水平均明显升高,差异有统计学意义(P<0.05);免疫荧光示p-STAT1阳性,主要位于肾小管上皮细胞中。结论 RACK1在糖尿病肾小管中高表达,可能是通过磷酸化STAT1激活Jak-STAT信号通路参与糖尿病肾小管病的发生发展。
基金supported by the National Natural Science Foundation of China (Grant No. 30571120)the National High Technology Research and Development Program of China (Grant No.2008AA10Z120)the Research Fund for the Doctoral Program of Higher Education, China
文摘The receptor for activated C-kinase 1 （RACK1） is a highly conserved scaffold protein with versatile functions, and plays important roles in the regulation of plant growth and development. Transgenic rice plants, in which the expression of RACK1 gene was inhibited by RNA interference （RNAi）, were studied to elucidate the possible functions of RACK1 in responses to drought stress in rice. Real-time PCR analysis showed that the expression of RACK1 in transgenic rice plants was inhibited by more than 50%. The tolerance to drought stress of the transgenic rice plants was higher as compared with the non-transgenic rice plants. The peroxidation of membrane and the production of malondialdehyde were significantly lower and the superoxide dismutase activity in transgenic rice plants was significantly higher than those in non-trangenic rice plants It is suggested that RACK1 negatively regulated the redox system-related tolerance to drought stress of rice plants.
