摘要
目的研究人绒毛滋养层细胞中调节细胞外基质金属蛋白酶诱导因子(EMMPRIN)表达的信号通路及p38MAPK信号通路在滋养细胞体外侵袭中的作用。方法体外无血清培养人绒毛滋养细胞,分别加入p38MAPK抑制剂(SB203580),JNK抑制剂(SP600125),ERK抑制剂(PD098059),用RT-PCR及Western blot方法观察阻断剂对EMM-PRIN表达的影响。用不同浓度的佛波酯(PMA)作用于滋养细胞,ELISA方法检测滋养细胞中p38MAPK的活性变化,用transwell细胞侵入系统检测滋养细胞的侵袭作用;加入不同浓度的SB203580,观察阻断剂对滋养细胞侵袭性的影响。结果JNK抑制剂、ERK抑制剂对滋养细胞分泌EMMPRIN无影响,p38MAPK抑制剂以时间剂量依赖的方式抑制滋养细胞表达EMMPRIN,SB203580浓度为5、10、15及20μmol/L作用24h后,EMMPIRN的抑制率分别为7.3%、24.6%、31.8%及39%;加入10μmol/L的SB203580培养24h后即可抑制EMMPRIN基因和蛋白的表达,抑制率为22%,培养48h和72h抑制率分别为45%和76%。向培养的细胞中加入浓度为0.1、1、10mmol/L的PMA作用30min,PMA以时间剂量依赖的方式激活p38MAPK,而SB203580以时间剂量依赖的方式抑制PMA对p38MAPK的激活。PMA可以促进滋养细胞体外侵袭作用,5mmol/L的SB203580能明显的抑制滋养细胞的体外侵袭能力,也能抑制PMA对滋养细胞侵袭活性的激活。结论p38MAPK信号传导途径参与了人绒毛滋养细胞中EMMPRIN的表达。p38MAPK通路在人滋养细胞的侵袭行为中有重要的作用,p38MAPK激动剂可能会为子痫前期-子痫的防治提供新的途径。
Objective: To study the effect on the in vitro invasion of cytotrophoblast by both EMMPRIN expression -regulating signal path and p38MAPK path in human Villus cytotrophoblast. Methods: Cultivate in vitro serum -free human villus cytotrophoblast, add p38MAPK inhibitor ( SB203580), JNK inhibitor ( SP600125 ), and ERK inhibitor ( PD098059 ) respectively, observe the irdluence of blocker on EMMPRIN expression through RT - PCR and Western blot method. Act on cytotrophoblast with phorbol - 12 - myris -tate - 13 -acetate (PMA) of different concentrations, detect the activity change of p38MAPK in cytotrophoblast through ELISA method, detect the invasion effect of cytotrophoblast through transwell cell invasion system; add SB203580 of different concentrations, and observe the influence of blocker on the invasion of cytotrophoblast. Results : JNK inhibitor and ERK inhibitor did not influence the EMMPRIN secretion of cytotrophoblast, but p38MAPK inhibitor inhibited the EMMPRIN expression of cytotrophoblast at in a dose - time- dependent manner. 24h after adding SB203580 at a concentration of 5, 10, 15, and 20μmol/L, the EMMPRIN inhibition rate was 7.3%, 24.6%, 31.8% , and 39% respectively. Add 10μmol/L SB203580 and at 24h after inhibitor inhibited the EMM- PRIN expression, the EMMPRIN inhibition rate was 22%, 45% and 76% respectively. 30min after adding PMA (at a concentration of 0. 1, 1, and 10mmol/L) into the cultured cell, PMA activated p38MAPK in a dose - time - dependent manner, but SB203580 inhibited such activation in a dose - time - dependent manner. PMA could promote the in vitro invasion effect of cytotrophoblast, but 5mmol/L SB203580 could significantly inhibited both such invasion and the activation of PMA on such invasion. Conclusion: p38MAPK signal conduction path participated in EMMPRIN expression in human villus cytotrophoblast. P38MAPK path had an important role in the invasion behavior of human cytotrophoblast, p38MAPK activator might provide new way to prevent and treat pre -eclampsia and eclampsia.
出处
《中国优生与遗传杂志》
2007年第8期19-22,共4页
Chinese Journal of Birth Health & Heredity