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目的:通过研究耳聋左慈丸对老年性耳聋小鼠耳蜗组织cAMP含量及PKA表达的影响,探讨老年性耳聋的治疗机制。方法:选取75只耳廓反应灵敏的雄性清洁级昆明小鼠,随机抽取15只作为空白对照组,腹腔注射生理盐水。剩余60只进行老年性耳聋小鼠造模,腹腔注射D-半乳糖(150 mg/kg),每日1次,共60 d。造模成功后将老年性耳聋模型小鼠随机分为模型组、耳聋左慈丸低、中、高剂量组。空白对照组和模型组给予生理盐水灌服,耳聋左慈丸低、中、高剂量组给予耳聋左慈丸(0.3、0.6、1.2 g/kg)灌服,每日1次,共14 d。运用耳蜗基底膜铺片技术观察小鼠耳蜗毛细胞形态学变化;采用ELISA法检测耳蜗组织中cAMP含量;用免疫组织化学技术检测PKA的蛋白表达水平;用RT-PCR技术检测PKA mRNA表达水平。结果:与空白对照组相比,模型组小鼠耳蜗基底膜毛细胞明显缺失或散乱,cAMP含量及PKA蛋白和mRNA表达明显下降;与模型组相比,经过耳聋左慈丸治疗的小鼠耳蜗毛细胞排列较整齐,cAMP含量及PKA蛋白和mRNA表达升高,其中高剂量组变化最为明显。结论:耳聋左慈丸可以通过调节耳蜗组织中cAMP含量及PKA的表达水平发挥其对老年性耳聋的治疗作用。
Objective: To study the effect of deafness Zuo Ci Pill on cAMP content and PKA expression in cochlear of aged mice with deafness, and to explore the therapeutic mechanism of senile deafness. Methods: Seventy five male Kunming mice with sensitive auricles were selected. Fifteen rabbits were randomly selected as control group and injected with normal saline intraperitoneally. The remaining 60 adult mice with deafness were induced by intraperitoneal injection of D-galactose (150 mg / kg) once daily for 60 days. After the success of modeling, the senile deafness model mice were randomly divided into model group, deafness Zuo Ci pill low, middle and high dose group. The rats in the control group and model group were fed with normal saline. The deafness Zuo Ci pill was given to low, middle and high dose deafness Zuo Ci Pills (0.3,0.6,1.2 g / kg) once a day for 14 days. The cochlear hair cell morphological changes were observed by cochlear basement membrane plating technique. The cAMP content in the cochlea was detected by ELISA. The protein expression of PKA was detected by immunohistochemistry. The expression of PKA mRNA was detected by RT-PCR. Results: Compared with the blank control group, the cochlear hair cells in the model group were significantly deleted or scattered hair cells, cAMP content and PKA protein and mRNA expression was significantly decreased; compared with the model group, mice treated with deafness Zuo Ci pill cochlea Hair cells arranged neatly, cAMP content and PKA protein and mRNA expression increased, of which the most obvious changes in high-dose group. CONCLUSION: Deafness Zuo Ci Pill can exert its therapeutic effect on senile deafness by regulating cAMP content and PKA expression level in cochlea.