p38丝裂原活化蛋白激酶通路对紫杉醇诱发细胞凋亡中γ-氨基丁酸B型受体表达变化的影响  

Effects of p38 mitogen-activated protein kinase pathway on changes of γ-aminobutyric acid B receptors expression in paclitaxel-induced apoptosis

作　　者：金子[1] 王秀丽[1] 吴川[1] 赵爽[1] 李昭[1] 柴潇潇 

机构地区：[1]河北医科大学第三医院麻醉科,石家庄050051

出　　处：《中华实验外科杂志》2015年第3期546-549,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30772082）;河北省自然科学基金资助项目（H2013206087）;河北省重点学科跟踪项目（GL2014041）

摘　　要：目的 通过给予p38丝裂原活化蛋白激酶（p38MAPKs）抑制剂（SB203580）,探讨紫杉醇诱发细胞凋亡中p38MAPKs通路对γ-氨基丁酸B型受体（GABAB受体）表达变化的影响及其作用机制.方法 随机选取原代培养5d,浓度约为1×109/L的海马细胞,给予不同浓度（0、0.01、0.10、1.00、10.00 μmol/L）紫杉醇,采用噻唑蓝（MTT）法及流式细胞仪检测法分别检测海马神经元细胞抑制率（n=3）及凋亡率（n=4）变化,以此确定紫杉醇诱发细胞凋亡的最适浓度.另选取原代培养5d的海马细胞随机分为4组：对照组（C组）、10 μmol/L SB203580处理组（K组）、紫杉醇最适浓度处理组（N组）、10 μmol/L SB203580＋紫杉醇最适浓度混合组（K＋N组）,培养时间为24h,保持各组加液量一致,Western blot法测定海马细胞GABAB受体及核因子-κB（NF-κB）蛋白表达水平（n=3）.结果 紫杉醇对海马细胞活力的抑制具有时间和浓度的交互作用（F=9.127,P＜0.05）,根据MTT法（n=3）和流式细胞仪法（n=4）检测结果,各浓度紫杉醇处理24h对海马神经元凋亡率的影响差异有统计学意义（F=64.523,P＜0.05）,并计算出紫杉醇诱发细胞凋亡的最适浓度为1 μmol/L[早期凋亡率达（48.63±5.76）％].蛋白表达结果显示（n=3）：与C组比较,N组和K＋N组GABAB受体与NF-κB表达均明显增高（P＜0.05）,而K组GABAB受体与NF-κB表达均降低（P＜0.05）;与K组比较,N组和K＋N组NF-κB和GABAB受体表达均增高（P＜0.05）;与N组比较,K＋N组NF-κB和GABAB受体表达均降低（P＜0.05）.结论 在紫杉醇诱发细胞凋亡过程中,抑制p38MAPKs通路可上调GABAB受体表达,而其下游因子NF-κB可能是其调控GABAB受体表达变化的关键靶点.Objective To investigate the effects of p38 mitogen-activated protein kinase （p38MAPK） pathway on changes of γ-aminobutyric acid B receptors （GABAs receptors） expression in paclitaxel-induced apoptosis by giving p38MAPK inhibitor （SB203580） and the and molecular mechanism.Methods The primarily cultured hippocampal neurons which had been cultivated for 5 days in vitro were randomly selected,and the density was about 1 × 109/L.These neurons were separately given different concentrations of paclitaxel （0,0.01,0.10,1.00,10.00 μmol/L）.The changes of hippocampal neuronral inhibitory rate （n =3） and apoptosis rate （n =4） were detected by methyl thiazol tetrazolium （MTT） method and flow cytometry.As a result,the optimal concentration of paclitaxel to induce apoptosis was determined.Hippocampal neurons were cultured for other 5 days,selected,and randomly divided into four groups：control group （group C）,10 μmol/L SB203580 group （K group）,the optimal concentration of paclitaxel group （N）,10 μmol/L SB203580 ＋ the optimal concentration of paclitaxel group （K ＋ N group）.All of the hippocampal neurons were cultured for 24 h,and the medicine volume in each group was consistent.The GABAB receptors and nuclear factor-κB （NF-κB） expression in the hippocampal neurons was detected by Western blotting （n =3）.Results Effect of paclitaxel on the viability of hippocampal neurons had duration and concentration interaction （F =6.127,P 〈 0.05）.According to the results of MTT method （n =3） and flow cytometry （n =4）,the apoptosis rate had a significant difference among each group （F =64.523,P 〈 0.05）,and then calculated the optimal concentration of paclitaxel-induced apoptosis was 1 μmol/L[The early apoptosis rate was （48.63 ± 5.76）%].As compared with group C （n =3）,the expression of GABAB receptors and NF-κB protein was significantly up-regulated in both N group and K ＋ N group （P 〈0.05）,and that of GABAB receptors and NF-κB protein was d

关 键 词：紫杉醇 P38丝裂原活化蛋白激酶 γ-氨基丁酸B型受体 核因子-ΚB 

分 类 号：R402[医药卫生—临床医学]