摘要
目的研究高糖对肾小球系膜细胞间隙连接蛋白(connexin43)表达和细胞间通讯功能的影响。方法分离培养大鼠肾小球系膜细胞,调整培养液葡萄糖浓度为以下3组:正常葡萄糖组(5.5mmol/L葡萄糖)、高糖组(30mmol/L葡萄糖)和渗透压对照组(5.5mmol/L葡萄糖加24.5mmol/L甘露醇),于37℃5%CO2条件下培养24、48h后,利用激光共聚焦显微镜和荧光漂白恢复(FRAP)技术检测细胞间通讯功能,并应用Northern印迹和细胞免疫化学、Western印迹方法检测connexin43mRNA和蛋白质表达,比较3组之间的差异。结果正常葡萄糖浓度培养下系膜细胞表达丰富的connexin43,细胞间通讯功能良好。高糖培养的系膜细胞细胞间通讯功能下降,荧光淬灭后的恢复比例和速度显著低于正常糖组(P<0.05)。同时高糖环境下培养的系膜细胞connexin.43mRNA和蛋白质表达均较正常糖组显著下降(P<0.05)。渗透压对照组与正常糖组之间差异无统计学意义(P>0.05)。结论高糖可抑制connexin43的基因和蛋白质表达及细胞间通讯功能,可能是糖尿病肾病系膜细胞表型和功能异常的重要原因之一。
Objective To investigate the influence of high glucose on connexin 43 expression and the gap junction intercellular communication （GJIC） function in mesangial cells. Methods The rat mesangial cells in vitro for 24 h or 48 h and were divided into normal glucose group （5.5 mmol/L）, high glucose group（30 mmol/L） and osmotic control group （5.5 mmol/L glucose plus 24.5 mmol/L mannitol）. Their GJIC function was measured by using fluorescence recovery after photobleaching assay （FRAP）. The gene and protein expression of connexin 43 was detected by Northern blot, immunocytochemistry and Western blot. Results Normal glucose group expressed rich connexin 43 and kept a good intercellular communication, whereas intercellular communication decreased in high glucose group. The fluorescence recovery rate and recovery period of highglucose group were significantly decreased compared to those of normal glucose group. The mRNA and protein expression of connexin 43 was reduced in high glucose group as compared to those in normal group. Conclusion High glucose can inhibit the GJIC function by reducing connexin 43 expression in rat mesangial cells, which may be responsible for the dysfunction and abnormality of mesangial cells in diabetic nephropathy.
出处
《中华肾脏病杂志》
CAS
CSCD
北大核心
2005年第8期443-447,共5页
Chinese Journal of Nephrology
基金
国家973资助项目(G2000057000)国家自然科学基金"创新研究群体"项目(30121005)国家自然科学基金资助项目(30300161)北京市自然科学基金资助项目(7032045)
