红景天苷改善顺铂引起的小鼠耳蜗毛细胞和螺旋神经节神经元损伤的机制
Sal idroside Modulates cAMP/PKA/CREB Signaling Pathway to Inhibit Ci splatin-induced Damages of Cochlear Hair Cells and Spiral Ganglion Neurons in Mice
李兆龙 1徐义策 1李泽文 1周洁1
作者信息
- 1. 孝感市中心医院耳鼻咽喉头颈外科(孝感 432000)
- 折叠
摘要
目的 探究红景天苷(SAL)改善顺铂(CIS)引起的耳蜗毛细胞(CHC)和螺旋神经节神经元(SGN)损伤的作用及其与环磷腺苷(cAMP)/蛋白激酶A(PKA)/cAMP效应元件结合蛋白(CREB)通路的关系.方法 分离新生C57BL/6小鼠的耳蜗基底膜,分为对照组(C组)、CIS组、SAL组、SAL+SQ22536(cAMP抑制剂)组和SAL+H-89(PKA抑制剂)组,每组20条.C组仅加入无血清BME培养液;CIS组在培养液中加入15 μmol/L CIS;SAL组在CIS组基础上加入5 μmol/L SAL;SAL+SQ22536组在CIS组基础上加入5 μmol/L SAL和5μmol/L SQ22536;SAL+H-89组在CIS组基础上加入5μmol/L SAL和30 μmol/L H-89.各组在培养箱中孵育48 h后,免疫荧光染色观察各组CHC和SGN损伤;试剂盒检测各组耳蜗基底膜中ROS和cAMP含量;Western blot检测各组PKA、p-CREB、CREB、Bcl-2、BDNF、NF-M蛋白水平.结果 CIS组CHC排列混乱、体积肿大,SGN细胞核破碎、神经突缺失,SAL可减轻CHC和SGNs损伤.与C组相比,CIS组CHC、SGN数量较少(P＜0.05),ROS、cAMP 含量、PKA、BDNF、NF-M、Bcl-2 蛋白及 p-CREB/CREB 水平较高(P＜0.05);与 CIS 组相比,SAL 组CHC、SGN 数量较多(P＜0.05),ROS 含量较低(P＜0.05),cAMP 含量、PKA、BDNF、NF-M、Bcl-2 蛋白及 p-CREB/CREB水平较高(P＜0.05).SQ22536和H-89均可逆转SAL对CHC和SGN的保护作用.结论 SAL可能通过激活cAMP/PKA/CREB通路,促进抗凋亡蛋白和神经保护因子表达,缓解CIS引起的CHC和SGN损伤.
Abstract
Objective To investigate the ameliorating effect of salidroside(SAL)on cisplatin(CIS)-induced damages of cochlear hair cells(CHC)and spiral ganglion neurons(SGNs)and its relationship with cyclic adenosine monophosphate(cAMP)/protein kinase A(PKA)/cAMP response element binding protein(CREB)pathway.Methods The cochlear basilar membranes of newborn C 57BL/6 mice were isolated and separated into control(C)group,CIS group,SAL group,SAL+SQ22536(cAMP inhibitor)group and SAL+H-89(PKA inhibitor)group,20 per group.Immunofluorescence staining was applied to observe the damages of CHC and SGNs.The kits were applied to detect the contents of ROS and cAMP in the basement membrane of the cochlea.Western blot was applied to detect the protein levels of PKA,p-CREB,CREB,Bcl-2,BDNF,and NF-M.Results CHC in CIS group were disorderly arranged and enlarged in size,SGNs had fragmented nuclei and lost neurites.SAL alleviated the damages of CHC and SGNs.Compared with the C group,the numbers of CHC and SGNs in the CIS group were less(P＜0.05),the contents of ROS and cAMP,and the levels of PKA,BDNF,NF-M,Bcl-2 proteins and p-CREB/CREB were higher(P＜0.05).Compared with the CIS group,the numbers of CHC and SGNs in the SAL group were higher(P＜0.05),the content of ROS was lower(P＜0.05),the content of cAMP,and the levels of PKA,BD-NF,NF-M,Bcl-2 proteins and p-CREB/CREB were higher(P＜0.05).Both SQ22536 and H-89 reversed the pro-tective effects of SAL on CHC and SGNs.Conclusion SAL may promote the expression of anti-apoptotic proteins and neuroprotective factors by activating the cAMP/PKA/CREB pathway to alleviate the damages of CHC and SGNs caused by CIS.
关键词
红景天苷/顺铂/毛细胞/螺旋神经节神经元/环磷腺苷/蛋白激酶A/cAMP效应元件结合蛋白通路Key words
Salidroside/Cisplatin/Hair cells/Spiral ganglion neurons/Cyclic adenosine monophos-phate/protein kinase A/cAMP response element-binding protein pathway引用本文复制引用
基金项目
孝感市自然科学基金(孝科技发[2021]3号)
出版年
2024
NETL
NSTL
维普
万方数据