摘要
目的研究D-半乳糖(D-gal)对小鼠TM4睾丸支持细胞屏障功能的损伤作用及机制。方法TM4细胞分为正常对照组和(25、50、100、150、200、250)mmol/L D-gal刺激组,噻唑蓝(MTT)法检测TM4细胞增殖活性,Western blot法检测TM4细胞紧密连接相关蛋白闭锁小带蛋白1(ZO-1)和闭合蛋白(occludin),黏附连接相关蛋白神经钙黏蛋白(N-cadherin)、上皮钙黏蛋白(E-cadherin)和β联蛋白(β-catenin),缝隙连接相关蛋白连接子蛋白43(CX43),骨架蛋白波形蛋白(vimentin)和丝裂原激活蛋白激酶(MAPK)信号通路相关蛋白胞外信号调节激酶1/2(ERK1/2)、Janus激酶(JNK)和p38MAPK的蛋白表达及磷酸化水平。结果与正常对照组比较,D-gal浓度大于50 mmol/L时,细胞活力显著下降;ZO-1、occludin、N-cadherin、E-cadherin和β-catenin的蛋白表达水平均显著降低,p38MAPK蛋白磷酸化水平显著增加,而CX43、vimentin及ERK1/2、JNK蛋白及磷酸化水平无明显变化。结论D-gal可引起TM4睾丸支持细胞紧密连接损伤和黏附连接损伤,可能与激活p38MAPK通路有关。
Objective To explore the effects of D-galactose(D-gal)on barrier function of murine TM4 sertoli cells and its mechanism.Methods TM4 cells were divided into control group and 25,50,100,150,200,250 mmol/L D-gal stimulation group.The viability of TM4 cells was determined by MTT assay.The protein expression levels of tight junction-related proteins including zonula occluden-1(ZO-1)and occludin,adheren junction-related proteins including neural cadherin(N-cadherin),epithelial cadherin(E-cadherin)andβ-catenin,gap junction-related protein connexin43(CX43)and cytoskeleton-related protein vimentin,and MAPK signaling pathway-related proteins ERK1/2,phosphorylated ERK1/2(p-ERK1/2),JNK,phosphorylated JNK(p-JNK),p38MAPK and phosphorylated p38MAPK(p-p38MAPK)were detected by Western blot analysis.Results Compared with the control group,the viability of TM4 cells significantly decreased when the concentration of D-gal was more than 50 mmol/L.In addition,the protein expression levels of ZO-1,occludin,N-cadherin,E-cadherin andβ-catenin were significantly down-regulated in D-gal-treated group,while the protein expression levels of p-p38MAPK were significantly up-regulated.However,there were no differences in the protein expression levels of CX43,vimentin,p-ERK,ERK1/2,p-JNK and JNK between the control group and D-gal-treated groups.Conclusion D-gal can disrupt tight junction and adheren junction of TM4 cells via the activation of p38MAPK signaling pathway.
作者
杨圆
张长城
张艳
杨思琪
叶勇
吴杰
袁丁
赵海霞
YANG Yuan;ZHANG Changcheng;ZHANG Yan;YANG Siqi;YE Yong;WU Jie;YUAN Ding;ZHAO Haixia(Third-Grade Pharmacological Laboratory on Traditional Chinese Medicine Approved by State Administration of Traditional Chinese Medicine,Medical College of China Three Gorges University,Yichang 443002,China)
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2020年第11期1002-1008,共7页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金(81774316,8187307,81573931)。
