摘要
该研究通过检测不同月龄C57BL/6J小鼠血管纹上S1PR1的表达情况,进一步探究老年耳蜗血管纹内皮细胞上S1PR1变化对血迷路屏障通透性的影响。用听性脑干反应(auditory brain response,ABR)检测听觉功能;伊文思蓝染色检测耳蜗血迷路屏障的通透性;免疫荧光检测耳蜗血管纹上S1PR1和紧密连接蛋白表达水平;ELISA检测S1P的释放量;Transwell小室检测内皮细胞通透性;Western blot检测EC上S1PR1和紧密连接蛋白的表达水平。结果显示,12m组小鼠听力阈值明显增高且高频听力损失明显,耳蜗血迷路屏障通透性升高,S1PR1和紧密连接蛋白表达水平均降低。在细胞水平,D-gal+EC组中S1P的释放量降低,S1PR1表达水平下降,加入外源S1P后,S1PR1表达水平增加;D-gal+EC组紧密连接蛋白表达水平降低,内皮细胞通透性增加,加入外源S1P后,紧密连接蛋白表达水平增加,内皮细胞通透性降低,给予S1PR1抑制剂干预后,紧密连接蛋白表达水平降低,内皮细胞通透性增加。在衰老状态下,耳蜗血管纹内皮细胞上S1PR1表达下降,这可能会下调紧密连接蛋白的表达,进而影响血迷路屏障通透性。
In this study,the paper examined the expression of S1PR1 in stria vascularis of C57BL/6J mice at different ages to explore the effect of S1PR1 on the permeability of blood-labyrinth barrier in endothelial cells of stria vascularis of the aged cochlea.ABR(auditory brain response)was used to measure auditory function.Evans blue staining for the permeability of the cochlear blood-labyrinth barrier.The expression of S1PR1 and connexin in stria vascularis of cochlea was detected by immunofluorescence.The release of S1P was detected by ELISA.The permeability of endothelial cells was measured by Transwell chamber.The expression levels of S1PR1 and tight junction protein in EC were detected by Western blot.The results showed that the hearing threshold and high frequency hearing loss were significantly increased in the 12m group,the permeability of cochlear blood-labyrinth barrier was increased,and the expression levels of S1PR1 and tight junction protein were decreased.At the cellular level,the release of S1P and the expression of S1PR1 were decreased in the D-gal+EC group,and S1PR1 expression was increased after the addition of exogenous S1P,while the expression of tight junction protein was decreased and endothelial permeability was increased in the D-gal+EC group,when exogenous S1P was added,tight junction protein expression was increased and endothelial permeability was decreased.When S1PR1 inhibitor was given,tight junction protein expression was decreased and endothelial permeability was increased.The decrease of S1PR1 expression in vascular stria endothelial cells of the cochlea during senescence may down-regulate the expression of tight junction protein,and then affect the permeability of blood labyrinth barrier.
作者
余蒙
余苗
孙诗恒
王皓阳
曾宪思
刘志丹
姜文君
李丽
YU Meng;YU Miao;SUN Shiheng;WANG Haoyang;ZENG Xiansi;LIU Zhidan;JIANG Wenjun;LI Li(School of Basic Medical Science,Zhejiang Chinese Medical University,Hangzhou 310000,China;College of Basic Medicine,Jiaxing University College of Medicine,Jiaxing 314000,China;Department of Physiology,Shihezi University College Xinjiang,Shihezi 832002,China)
出处
《中国细胞生物学学报》
CAS
CSCD
2023年第8期1141-1151,共11页
Chinese Journal of Cell Biology
基金
国家自然科学基金(批准号:81960188)
浙江省政府自然科学基金(批准号:LY21H130001)资助的课题。
