| Objective: to verify the therapeutic methods of acupuncture and acupoint injection through experimental studies,which can reduce the damage of cochlea induced by GM in mice with ototoxicity,so as to protect the morphological integrity of cochlea hair cells and spiral ganglion and reduce the ABR threshold.It also promoted the synthesis of antioxidant enzymes in the cochlea tissue and improved the ability of the inner ear to resist oxidative damage.It affects the gene transcription and protein expression of NrF2 signaling pathway,and antagonizes the effect of gentamicin-induced apoptosis of hair cells and ganglion cells.Methods:The experiment was divided into five groups,and the pre-experiment grouping,shaping,drug administration,treatment and brainstem potential test were all the same in each group.48 normal healthy kunming mice(96 ears)were randomly divided into 6 groups,with 8 mice in each group:Ⅰ.Negative control group: from day 1 to 10,daily intraperitoneal injection of normal saline,2.5ml /Kg.Ⅱ.Model group(GM): daily intraperitoneal injection of GM,100mg/kg and intraperitoneal injection of GM,100mg/kg were performed on days 1 to 10.Ⅲ.Positive drug group(GM+ EGB): from day 1 to 10,GM+ EGB was injected intraperitoneally at 8:00 and 16:00,7mg/kg each.Ⅳ,acupuncture group(GM+ acupuncture): from day 1 to 10,GM was injected intraperitoneally at 8:30,100mg/kg daily.From the 2nd day to the 10 th day,take the hearing palace,listen to the meeting,yifeng acupuncture,acupuncture for 10 minutes.Ⅴ.Acupoint injection group(GM+ acupoint injection): from day 1 to 10,GM was injected intraperitoneally at 8:30,100mg/kg daily.From the 2nd day to the 10 th day to take the hearing palace,yi feng,tip of the ear acupuncture point injection.Ⅵ.Acupuncture + acupoint injection group(GM+ acupuncture + acupoint injection): from day 1 to 10,GM was injected intraperitoneally at 8:30,100mg/kg daily.From the 2nd day to the 10 th day,take the hearing palace,listen to the meeting,yifeng acupuncture,acupuncture for 10 minutes.Yu heigong,heighui,yi feng acupuncture point injection.ABR detection was performed in each group on the first day after drug withdrawal,and the threshold shift of ABR was compared with that of the model group.The effects of acupuncture and acupoint injection on the hearing threshold of gentamicin-induced deafness mice were analyzed.Different extraction methods were used to extract the cochlea tissue according to the different requirements on the cochlea tissue in different experiments.Changes in the morphology and number of cochlea hair cells were observed,changes in the antioxidant system of the cochlea tissue were analyzed,changes in the transcription level of NRF2 signaling pathway genes were observed,and differences in the expression of proteins related to the NRF2 signaling pathway were compared.Result: 1.Model group testing results of ABR threshold shift was 17.50 + /-2.43,compared with negative control group plus or minus 1.67 1.6,P < 0.001,significant difference,statistically significant,proved successful gentamycin induced deafness model is established,and the positive control group,acupuncture group,acupuncture point injection group,the acupuncture plus point injection group all have different degrees of threshold shift,lower than the model group,acupuncture plus point injection group compared with model group significant difference.2.In the negative control group,the inner and outer hair cells in the cochlea were arranged neatly,while in the model group,the inner and outer hair cells in the cochlea were all lost and not arranged neatly.The loss of hair cells in the positive control group,acupuncture group,acupoint injection group and acupuncture plus acupoint injection group all varied,and the arrangement tended to be orderly.The hair cell loss in the acupuncture plus acupoint injection group was the least,that is,the treatment effect was the best.3.In the experiment of anti-oxidative stress,the activity of SOD and gsh-px increased and the activity of gsh-px decreased after acupuncture and acupoint injection.4.After acupuncture and acupoint injection,the expressions of Nrf2,ho-1and NQO1 in the cochlea tissue were detected to be increased.5.In the immunohistochemical experiment,the expression of HO1,nqo-1 and Nrf2 in HC and SGN increased after acupuncture and acupoint injection,while the expression of caspase-3 decreased.In the immunohistochemical photos,HO1,nqo-1and Nrf2 in HC and SGN groups showed higher positive expression in the acupuncture plus acupoint injection group,while the caspase-3 group showed lower positive expression in the acupuncture plus acupoint injection group.Conclusion:1.GM can induce ototoxicity and increase the ABR threshold of mice;2.Destroy hair cell,spiral ganglion shape,induce cell death.2.The treatment of acupuncture group and acupoint injection group can reduce the ototoxicity of GM,reduce the damage of GM to cochlea hair cells and spiral ganglion,protect the morphological integrity of hair cells and spiral ganglion,and reduce the ABR threshold.Gentamicin damages the antioxidant system of cochlea tissue and inhibits the synthesis of antioxidant enzyme system.Acupuncture and acupoint injection can promote the synthesis of antioxidant enzymes and improve the ability of the inner ear to resist oxidative damage.4.Acupuncture and acupoint injection therapy had significant effects on gene transcription and protein expression of NrF2 signaling pathway in cochlear tissues of gentamicin-induced deafness mice,activated the mechanism of anti-oxidative stress,and antagonized the effect of gentamicin-induced apoptosis of hair cells and ganglion cells.5.Among the treatment groups,acupuncture + acupoint injection group had the best therapeutic effect. |
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