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

Salidroside Modulates cAMP/PKA/CREB Signaling Pathway to Inhibit Cisplatin-induced Damages of Cochlear Hair Cells and Spiral Ganglion Neurons in Mice

李兆龙;徐义策;李泽文;周洁

1:孝感市中心医院耳鼻咽喉头颈外科

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15μmol/L CIS;SAL组在CIS组基础上加入5μmol/L SAL;SAL+SQ22536组在CIS组基础上加入5μmol/L SAL和5μmol/L SQ22536;SAL+H-89组在CIS组基础上加入5μmol/L SAL和30μmol/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。
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
基金项目(Foundation):
孝感市自然科学计划项目(孝科技发[2021]3号)
作者
李兆龙;徐义策;李泽文;周洁
参考文献

1 Freyer DR,Brock PR,Chang KW,et al.Prevention of cisplatin-induced ototoxicity in children and adolescents with cancer:a clinical practice guideline[J].Lancet Child Adolesc Health,2020,4(2):141-150.

2 Tang Q,Wang X,Jin H,et al.Cisplatin-induced ototoxicity:updates on molecular mechanisms and otoprotective strategies[J].Eur J Pharm Biopharm,2021,163(1):60-71.

3 Ramkumar V,Mukherjea D,Dhukhwa A,et al.Oxidative stress and inflammation caused by cisplatin ototoxicity[J].Antioxidants (Basel),2021,10(12):1919.

4 Magani SKJ,Mupparthi SD,Gollapalli BP,et al.Salidroside—can it be a multifunctional drug[J]?Curr Drug Metab,2020,21(7):512-524.

5 于姝媛,李浩楠,朱紫嘉,等.红景天苷通过降低氧化应激介导氨基糖苷类药物耳毒性的抑制作用[J].中国听力语言康复科学杂志,2018,16(4):246-249.

6 Ding X,Wang W,Chen J,et al.Salidroside protects inner ear hair cells and spiral ganglion neurons from manganese exposure by regulating ROS levels and inhibiting apoptosis[J].Toxicol Lett,2019,310(1):51-60.

7 Basch ML,Brown RM,Jen HI,et al.Where hearing starts:the development of the mammalian cochlea[J].J Anat,2016,228(2):233-254.

8 Gao X,Zhang X,Cui L,et al.Ginsenoside Rb1 promotes motor functional recovery and axonal regeneration in post-stroke mice through cAMP/PKA/CREB signaling pathway[J].Brain Res Bull,2020,154(1):51-60.

9 Yang Y,Ma S,Wei F,et al.Pivotal role of cAMP-PKA-CREB signaling pathway in manganese-induced neurotoxicity in PC12 cells[J].Environ Toxicol,2019,34(9):1052-1062.

10 Mei H,Zhao L,Li W,et al.Inhibition of ferroptosis protects house ear institute-organ of corti 1 cells and cochlear hair cells from cisplatin-induced ototoxicity[J].J Cell Mol Med,2020,24(20):12065-12081.

11 Kim YJ,Lee JS,Kim H,et al.Gap junction-mediated intercellular communication of cAMP prevents CDDP-induced ototoxicity via cAMP/PKA/CREB pathway[J].Int J Mol Sci,2021,22(12):6327-6337.

12 Breglio AM,Rusheen AE,Shide ED,et al.Cisplatin is retained in the cochlea indefinitely following chemotherapy[J].Nat Commun,2017,8(1):1654-1662.

13 Gentilin E,Simoni E,Candito M,et al.Cisplatin-induced ototoxicity:updates on molecular targets[J].Trends Mol Med,2019,25(12):1123-1132.

14 Zhang X,Zhang Y,Li R,et al.Salidroside ameliorates Parkinson's disease by inhibiting NLRP3-dependent pyroptosis[J].Aging (Albany NY),2020,12(10):9405-9426.

15 Hu M,Zhang D,Xu H,et al.Salidroside activates the AMP-activated protein kinase pathway to suppress nonalcoholic steatohepatitis in mice[J].Hepatology,2021,74(6):3056-3073.

16 Jewell JL,Fu V,Hong AW,et al.GPCR signaling inhibits mTORC1 via PKA phosphorylation of raptor[J].Elife,2019,8(1):e43038.

17 Sisakht M,Khoshdel Z,Mahmoodazdeh A,et al.Adrenomedullin increases cAMP accumulation and BDNF expression in rat DRG and spinal motor neurons[J].Iran J Basic Med Sci,2021,24(7):978-985.

18 Li S,Lu Y,Ding D,et al.Fibroblast growth factor 2 contributes to the effect of salidroside on dendritic and synaptic plasticity after cerebral ischemia/reperfusion injury[J].Aging (Albany NY),2020,12(11):10951-10968.

19 Yan J,Xu W,Lenahan C,et al.CCR5 activation promotes NLRP1-dependent neuronal pyroptosis via CCR5/PKA/CREB pathway after intracerebral hemorrhage[J].Stroke,2021,52(12):4021-4032.

20 Chen X,Wu Y,Yang T,et al.Salidroside alleviates cachexia symptoms in mouse models of cancer cachexia via activating mTOR signalling[J].J Cachexia Sarcopenia Muscle,2016,7(2):225-232.

21 冯晓玲,孙佳宁,赵小萱.红景天苷联合顺铂对人卵巢癌SKOV-3裸鼠移植瘤生长影响及其机制研究[J].中华中医药杂志,2021,36(7):3851-3855.