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目的探讨p38丝裂原活化蛋白激酶(p38MAPK)通路与细胞空泡形成的关系。方法应用茴香霉素、放线菌酮、p38MAPK抑制剂SB203580、JNK抑制剂SP600125处理HepG2、LM3、QBC939、Hela和A549细胞,光学显微镜和激光共聚焦显微镜观察细胞空泡化情况;Western blot法检测p38MAPK等通路相关分子的表达水平;内质网红色荧光探针标记内质网,激光共聚焦显微镜观察内质网结构变化;溶酶体红色荧光探针标记溶酶体,激光共聚焦显微镜观察溶酶体荧光染色情况。结果 (1)茴香霉素对HepG2细胞空泡有消除作用。(2)茴香霉素通过活化p38MAPK消除细胞空泡。(3)阻断p38MAPK诱导多种肿瘤细胞空泡形成。(4)阻断p38MAPK介导的空泡形成破坏内质网结构的整体性。(5)阻断p38MAPK介导的空泡形成具有可逆性。结论 p38MAPK通路在调节细胞空泡形成中发挥了重要作用。
Objective To investigate the relationship between the p38 mitogen-activated protein kinase (p38MAPK) pathway and the formation of cellular vacuoles. Methods HepG2, LM3, QBC939, Hela and A549 cells were treated with anisomycin, cycloheximide, p38MAPK inhibitor SB203580 and JNK inhibitor SP600125. The cell vacuolization was observed by light microscope and confocal laser scanning microscopy. p38MAPK and other pathways related to the expression of endoplasmic reticulum red fluorescent probe labeled endoplasmic reticulum, confocal laser scanning microscope changes in endoplasmic reticulum structure; lysosomal red fluorescent probe labeled lysosomes, laser scanning confocal microscope Enzyme fluorescence staining. Results (1) Anisomycin had no effect on the vacuolization of HepG2 cells. (2) Anisomycin eliminates cellular vacuoles by activating p38MAPK. (3) block p38MAPK induced a variety of tumor cells vacuolar formation. (4) block p38MAPK-mediated vacuolization disrupts the integrity of the endoplasmic reticulum structure. (5) Blocking p38MAPK-mediated vacuolar formation is reversible. Conclusion The p38 MAPK pathway plays an important role in the regulation of cell vacuolization.