Objective:To study the effects of Erlong Zuoci Pill(耳聋左慈丸,ELZCP)and its disassembled prescriptions on gentamicin(GM)-induced ototoxicity model in vitro.Methods:After the spiral organ of cochleae of newborn mice(postnatal days:2-3)cultured for 24 h,GM alone or combined with water extracting-alcohol precipitating solution of ELZCP or with its disassembled prescriptions was added.Hair cells were observed under a fluorescence microscope after TRITC-phalloidin staining,and the cochlear hair cell loss rate was calculated by counting the whole cochlear hair cells and analyzed by whole cochlear hair cells analyzing software.Results: GM induced cochlear outer hair cells(OHCs)and inner hair cells(IHCs)injuries in a dose-dependent manner, and they were significantly different as compared with those in the normal control group(P<0.05,P<0.01). ELZCP at the concentration of 0.003-3 mg/mL could decrease the hair cells loss induced by the 0.3 mmol/L GM (P<0.05,P<0.01),the effects was in a dose-dependent manner,and the concentration of 0.3 mg/mL showed the optimal protective effect.For the ELZCP disassembled prescriptions,Liuwei-Dihuang could decrease OHC loss rate than that in the 0.3 mmol/L GM model group(P<0.05),but the OHC loss rate was still higher than that in the ELZCP group(P<0.01),which indicated that the protective effect of hair cells by Liuwei-Dihuang was not better than that of ELZCP.Poria decreased OHC loss rate from 72.1%±3.7%to 58.8%±8.2%(P<0.05). Conclusions:ELZCP could play a role in antagonizing the injury of cochlear hair cells induced by GM ototoxicity, and its disassembled prescriptions,Liuwei-Dihuang was the main component to protect the cochlear hair cells from GM-induced ototoxicity,and Magnetitum combined with Radix Bupleurui could strengthen the action of the whole prescription;Poria could reduce GM-induced OHC loss. Objective: To study the effects of Erlong Zuoci Pill and its disassembled prescriptions on gentamicin (GM) -induced ototoxicity model in vitro. Methods: After the spiral organ of cochleae of newborn mice (postnatal days: 2 -3) cultured for 24 h, GM alone or combined with water extracting-alcohol precipitating solution of ELZCP or with its disassembled prescriptions was added. Hair cells were observed under a fluorescence microscope after TRITC-phalloidin staining, and the cochlear hair cell loss rate was calculated by counting the whole cochlear hair cells and analyzed by whole cochlear hair cells analyzing software. Results: GM induced cochlear outer hair cells (OHCs) and inner hair cells (IHCs) injuries in a dose-dependent manner, and they were significantly different ELZCP at the concentration of 0.003-3 mg / mL could decrease the hair cells loss induced by the 0.3 mmol / L GM (P <0.05, P <0.05, P <0.01) <0.01), the effects was in a do For the ELZCP disassembled prescriptions, Liuwei-Dihuang could decrease OHC loss rate than that in the 0.3 mmol / L GM model group (P <0.05), and the concentration of 0.3 mg / but the OHC loss rate was still higher than that in the ELZCP group (P <0.01), which indicated that the protective effect of hair cells by Liuwei-Dihuang was not better than that of ELZCP. Poria decreased OHC loss rate from 72.1% ± 3.7% to 58.8% ± 8.2% (P <0.05). Conclusions: ELZCP could play a role in antagonizing the injury of cochlear hair cells induced by GM ototoxicity, and its disassembled prescriptions, Liuwei-Dihuang was the main component to protect the cochlear hair cells from GM-induced ototoxicity, and Magnetitum combined with Radix Bupleurui could strengthen the action of the whole prescription; Poria could reduce GM-induced OHC loss.