摘要
目的探讨壳聚糖（cs）介导细胞因子反应调节蛋白A（CrmA）对软骨细胞IL-1β转化酶（ICE）和IL-1β mRNA和蛋白的表达作用。方法体外培养兔关节软骨细胞，分别加入PBS、10μg／mlCS／pcDNA3．1（＋）和cs／r,cDNA3．1（＋）CrmA处理后，加入10ng／ml IL-1β共培养，采用实时定量PCR方法检测软骨细胞ICE和IL-1β mRNA的表达。蛋白印迹法分析软骨细胞ICE和IL-1β蛋白的表达。结果ICEmRNA在Cs／pcDNA3．1（＋）CrmA组（0．52±0．09）软骨细胞中的表达显著低于CS／pcDNA3．1（＋）组（0．84±0．11，t=4．42，P〈0．01）和PBS组（1．00±0．10，t=6．58，P〈0．01），软骨细胞ICE蛋白在CS／pcDNA3．1（＋）CrmA组（0．20±0．03）的表达明显低于CS／pcDNA3．1（＋）组（0．37±0．05，f=4．85，P〈0．01）和PBS组（0．44±0．07，t=6．68，P〈0．01），cs／pcDNA3．1（＋）组软骨细胞ICEmRNA和蛋白的表达与PBS组比较差异无统计学意义。与CS／pcDNA3．1（＋）组（0．69±0．06，t=3．50，P〈0．01）和PBS（0．99±0．04，t=11．12，P〈0．01）组比较，cs／pcDNA3．1（＋）CrmA组软骨细胞IL-1β t=3．50，（0．55±0．08）mRNA表达明显降低。软骨细胞IL-1β蛋白的表达在CS／pcDNA3．1（＋）CrmA组（0．230±0．020）显著低于CS／pcDNA3．1（＋）组（0．450±0．060，t=5．07，P〈0．01）与PBS组（0．610±0．090，t=8．70，P〈0．01），cs／pcDNA3．1（＋）组与PBS组比较软骨细胞IL-1β mRNA和蛋白的表达差异有统计学意义（t=7．61，P〈0．01；f=3．63，P〈0．01）。结论壳聚糖介导CrmA能够显著抑制ICEmRNA和蛋白的表达，从而下调IL-1β mRNA和蛋白的表达，这可能成为CS／pcDNA3．1（＋）CrmA抑制OA软骨退变的作用机制之一。
Objective To investigate the effects of chitosan（CS）/pcDNA3.1（＋） CrmA on the expression of interleukin-1β （IL-1β） converting enzyme （ICE） and IL-1β in chondrocytes. Methods Rabbit chondrocytes were isolated and cultured. Chondrocytes were treated with PBS, 10 μg/ml CS/pcDNA3.1 （＋） and CS/pcDNA3.1 （＋） CrmA respectively for 6 hours. Then 10 ng/ml IL-1β was added into the culture medium. After 48 hours, the messenger RNA and protein expression of ICE and IL-1β in chondrocytes were detected by using real time polymerase chain reaction and western blotting. Results In CS/pcDNA3.1 （＋） CrmA treated group （0.52 ±0.09）, the mRNA expression of ICE inchondrocytes was significantly inhibited compared withcorresponding samples of CS/pcDNA3.1（＋） group （0.84±0.11, t=4.42, P〈0.01） and PBS group （1.00±0.10, t=6.58, P〈0.01）. ICE protein expression in CS/pcDNA3.1（＋） CrmA treated group （0.20±0.03） was markedly lower than CS/pcDNA3.1 （＋） （0.37±0.05, t=4.85, P〈0.01） and PBS treated group （0.44±0.07, t=6.68, P〈0.01）. There was no significant difference of ICE mRNA and protein expressions in chondrocytes between CS/pcDNA3.1（＋） group and PBS group. Significant difference of IL-1β mRNA expression was found in the three groups. IL-1β mRNA expression level was significantly lower in CS/peDNA3.1（＋） CrmA treated group （0.55± 0.08） than CS/pcDNA3.1（＋） （0.69±0.06, t=3.50, P〈0.01） and PBS group （0.99±0.04, t=11.12, P〈0.01）of chondrocytes. IL-1β protein expression level of chondrocytesin CS/pcDNA3.1（＋） CrmA group （0.230±0.020） was significantly lower than CS/peDNA3.1 （＋） （0.450±0.060, t=5.07, P〈0.01 ）and PBS groups （0.610±0.090, t=8.70, P〈0.01） of eells. Significant differenee of IL-1β mRNA and protein expression between CS/peDNA3.1 （＋） and PBS group was also observed （t=7.61, P〈0.01; t=3.63, P〈0.01）. Conclusion CrmA mediated by ehitosan ean signifieantly suppress t
出处
《中华风湿病学杂志》
CAS
CSCD
北大核心
2016年第9期609-613,共5页
Chinese Journal of Rheumatology
基金
国家自然科学基金（81071494）
中央高校基本科研业务费专项资金项目（2042015kf10-14）
湖北省科技支撑计划项目（2015BCA316）
湖北省卫生计生科研项目（WJ2015MB024）
