阿司匹林对顺铂损伤豚鼠耳蜗螺旋神经节HO-1表达的影响

Effect of aspirin on heme oxygenase-1 expression in the cochlear spiral ganglion of guinea pigs with cisplatin-induced ototoxicity

目的:研究阿司匹林(acetylsalicylic acid,ASA)对顺铂(cisplatin,DDP)损伤豚鼠耳蜗螺旋神经节(spiral ganglion,SGNs)细胞中血红素加氧酶-1(heme oxygenase-1,HO-1)表达的影响,并探讨ASA通过诱导HO-1的表达对DDP损伤豚鼠耳蜗螺旋神经节的保护作用。方法:将40只豚鼠随机分成4组,每组10只。A组(正常对照组):每日以0.9%生理盐水10 mL/kg腹腔注射;B组(DDP组):每日腹腔注射顺铂10 mg/kg;C组(ASA+DDP组):每日腹腔注射阿司匹林100 mg/kg预处理1 h,再腹腔注射顺铂10 mg/kg;D组[HO-1抑制剂锌原卟啉IX(ZnppIX)+ASA+DDP组]:先同时予以ZnppIX 10μmol/kg+阿司匹林100 mg/kg腹腔注射预处理1 h,再予以顺铂10 mg/kg腹腔注射。每组均早晚各一次,持续用药3 d后,观察各组豚鼠ABR阈值变化,并采用免疫组织学、免疫荧光学、蛋白免疫印迹检测各组间耳蜗螺旋神经节细胞中HO-1的表达情况。结果:在各组豚鼠的检测结果中,D组ABR阈值最高(47.18±2.56),其次是B组(39.98±7.32)、C组(2.85±2.38),A组最低(1.65±2.13);免疫组织学、免疫荧光学的HO-1检测显示A组>C组>B组>D组;Western blot检测结果显示,A组HO-1表达最多(0.98±0.07),其次是C组(0.63±0.02)、B组(0.27±0.05),D组最少(0.12±0.09),4组间差异有统计学意义(P=0.000)。结论:阿司匹林可以通过增加耳蜗螺旋神经节HO-1的表达,对顺铂诱导的SGNs损伤起保护作用。

Objective:To study the effect of aspirin(ASA) on the expression of heme oxygenase-1(HO-1) in cochlear spiral ganglion neurons(SGNs) of guinea pigs with cisplatin(DDP) ototoxicity,and to explore the protective effect of ASA on cochlear SGNs of guinea pigs with DDP ototoxicity by inducing the expression of HO-1. Methods:Forty guinea pigs were randomly and equally divided into 4 groups:group A(normal control group) received intraperitoneal injection of 0.9% normal saline at a daily dose of 10 mL/kg;group B(DDP group) received intraperitoneal injection of DDP at a daily dose of 10 mg/kg;group C(ASA+DDP group) received intraperitoneal injection of DDP at a daily dose of 10 mg/kg following 1-hour pretreatment of 100 mg/kg ASA;group D [HO-1 inhibitor zinc protoporphyrin IX(ZnppIX)+ASA+DDP group] received intraperitoneal injection of DDP at a daily dose of 10 mg/kg following 1-hour pretreatment of 10 μmol/kg ZnppIX and 100 mg/kg ASA. Each group received treatment once in the morning and once in the evening. After 10 days of continuous administration,changes in auditory brainstem response(ABR) threshold were observed in each group. The HO-1 expression in cochlear SGNs was measured by immunohistochemistry,immunofluorescence,and Western blot.Results:In all the four groups,group D had the highest ABR threshold(47.18±2.56),followed by group B(39.98±7.32),group C(2.85±2.38),and group A(1.65 ±2.13). The immunohistochemistry and immunofluorescence results showed that group A had the highest HO-1 expression,followed by group C,group B,and group D. The Western blot results showed that there were significant differences between the four groups in HO-1 expression(P=0.000) and group A had the highest expression(0.98 ±0.07),followed by group C(0.63±0.02),group B(0.27±0.05),and group D(0.12±0.09). Conclusion:ASA can protect SGNs from DDP-induced ototoxicity by increasing the expression of HO-1 in cochlear SGNs.