受体酪氨酸激酶Mer对高糖环境培养的大鼠雪旺细胞系RSC96增殖调控作用观察  

Role of receptor tyrosine kinase Mer in regulating proliferation of rat Schwann cell line RSC96 cultured in a high-glucose

作　　者：付怡丹 陈文婷 苏晓杨 赵燕[3] 兰丹凤 杨秋萍[5] FU Yidan;CHEN Wenting;SU Xiaoyang;ZHAO Yan;LAN Danfeng;YANG Qiuping(不详;Cadre Medical Department,The First Affiliated Hospital of Kunming Medical University,Kunming 650032,China)

机构地区：[1]昆明医科大学第一附属医院干部医疗科,昆明650032 [2]昆明医科大学第一附属医院重症医学科 [3]昆明医科大学第一附属医院糖尿病科 [4]云南省第一人民医院消化内科 [5]昆明医科大学第一附属医院内分泌一科

出　　处：《山东医药》2024年第6期29-33,共5页Shandong Medical Journal

基　　金：云南省科技厅科技计划项目(202001AY070001-159)。

摘　　要：目的观察受体酪氨酸激酶Mer(MER tyrosine kinase,Mertk)在高糖环境培养条件下的大鼠雪旺细胞系RSC96增殖的调控作用。方法取RSC96分为一、二、三、四、五组,分别置于含25(正常葡萄糖浓度)、50、75、100及125 mmol/L葡萄糖的培养基中培养,培养48 h时采用Western Blotting法检测各组细胞Mertk,筛选Mertk蛋白相对表达量最高浓度为后续研究的高糖浓度。取RSC96细胞先饥饿处理4 h,置入含100 mmol/L的葡萄糖培养基中,分别于培养0、24、36、48、60 h时取各组细胞,采用Western Blotting法检测各组细胞Mertk,最终筛选Mertk蛋白相对表达量高的时间为后续研究培养时间。取RSC96细胞,分为对照组、高糖组、高糖敲降组及敲降组:高糖组细胞饥饿处理4 h,置于含100 mmol/L葡萄糖的培养基中培养;高糖敲降组细胞饥饿处理4 h,置于含100 mmol/L葡萄糖的培养基中,后加入5μL的敲降Mertk基因表达siRNA溶液;敲降组细胞饥饿处理4 h,加入5μL的敲降Mertk基因表达siRNA溶液;对照组细胞用正常培养基培养。培养48 h时取各组细胞,采用Edu法测算各组细胞增殖活力,采用Western Blotting法检测各组细胞Mertk、磷酸化核转录因子κB、P65及肿瘤坏死因子α(TNF-α)。结果与一组相比,四、五组细胞Mertk蛋白相对表达量高(P均<0.05);与培养0 h相比,培养48、60 h时RSC96细胞Mertk相对表达量高(P均<0.05)。与对照组相比,高糖组细胞增殖活力低(P<0.05),敲降组细胞增殖活力高(P<0.05);与对照组相比,高糖敲降组细胞P65、TNF-α相对表达量高(P均<0.05),敲降组细胞Mertk相对表达量低、P65及TNF-α相对表达量高(P均<0.05),高糖组细胞Mertk、P65及TNF-α相对表达量高(P均<0.05)。结论敲降Mertk基因表达的高糖环境培养RSC96细胞的增殖能力高,细胞P65、TNF-α表达高。Mertk可能通过促进细胞P65、TNF-α表达,促进高糖环境培养RSC96细胞的增殖。Objective To observe the regulatory role of receptor tyrosine kinase Mer(Mertk)in the proliferation of the rat Schwann cell line RSC96 under high glucose condition.Methods RSC96 cells were divided into groups one,two,three,four and five and were cultured in medium containing 25(normal glucose concentration),50,75,100 and 125 mmol/L glucose,respectively,and Mertk was detected in cells of each group by Western blotting at 48 h of incuba‐tion,and the highest concentration of Mertk protein relative expression was selected to be the high-glucose concentration for the subsequent study.RSC96 cells were starved for 4 h,and were placed in the glucose medium containing 100 mmol/L glucose,and then were cultured for 0,24,36,48,and 60 h.Western blotting was used to detect the Mertk of cells in each group,and the time with the highest relative expression of Mertk protein was screened as the time of culture for the subsequent study.RSC96 cells were taken and divided into the control group,high glucose group,high glucose knock‐down group and knockdown group:cells in the high glucose group were starved for 4 h and cultured in medium containing 100 mmol/L glucose;cells in the high glucose knockdown group were starved for 4 h,and were placed in medium contain‐ing 100 mmol/L glucose,and then,5μL of siRNA solution with knockdown of Mertk expression was added;cells in the knockdown group were starved for 4 h,and 5μL of siRNA with knockdown of Mertk expression was added;cells in the control group were cultured in the normal medium.The cells in each group were taken at 48 h of culture,and the prolifera‐tion of each group was measured by Edu method,and Mertk,phosphorylated nuclear transcription factor-κB(P65)and tu‐mor necrosis factor-α(TNF-α)in each group were detected by Western blotting.Results Compared with the group one,the relative expression levels of Mertk protein were higher in cells of groups four and five(all P<0.05);the relative expres‐sion of Mertk was higher in RSC96 cells at 48 and 60 h of culture in compa

关 键 词：受体酪氨酸激酶 受体酪氨酸激酶Mer 葡萄糖 雪旺细胞 细胞增殖 核转录因子κB P65基因 肿瘤坏死因子α 

分 类 号：R587.11[医药卫生—内分泌]