The Effect Of "Erlong Zuoci Pill" On SOD?MDA?PKC In Mice Serum And Cochlea With Presbycusis Model

Purpose : To experiment in D-gal induced presbycusis in mice as model, based on the resisitance to free radical damage,based on the theory of “kidney main ear” and “From the kidney treatment deaf”,to observe the “Erlong Zuoci Pill”on serum and cochlea of mice with the effect of SOD ? MDA ? PKC,to investigate the treatment mechanism of “Erlong Zuoci Pill”to presbycusis,to provide scientific experimental basis for “Erlong Zuoci Pill”.Material and method:Sixty healthy male SPF Kunming mice were randomly divided into 2 groups: the control group had twelve mice, the molding group had forty-eight mice.The mice in control group were treated with 0.9% Na Cl and the other mice were treated with ip D-galactose( 150mg/kg.d) once a day, with 2 months. After duplicating the mold, the seventy-five mice were randomly divided into 4 groups:model group,Chinese medicine low dose group,Chinese medicine mid-dose group,Chinese medicine high dose group,with 12 mice in each group.The Chinese medicine group were treated with different density Erlong Zuoci Pill, included the low dose group(3g/Kg.d)?the mid-dose group(6g/Kg.d)?the high dose group(12g/Kg.d)once a day,with 14 days. We checked its listening ability by auditory brain-stem response( ABR) and used colorimetric method to determine the changes of the serum SOD activity and MDA content and the PKC in rats serum was detected by double antibody body clip ELISA method.Cochlea basilar membrane stretched preparation was used to observed the morphological changes of hair cells.Immunohistochemistry was used to detect the protein expression of PKC with the cochlear of the mice.RT-PCR was used to detect the gene expression of PKC ?SOD with the cochlear of the mice.Results:1.The general condition in mice:Compared with the control group, the general condition inmice of the molding group become worse;Compared with he molding group, three Chinese medicine treatment group were improved.2.The hearing test in mice(ABR): In model group,the value of ABR was markedly increased after the establishment of model,difference was statistical significance(P<0.01).Orally after weeks,compared with the control group, the value of ABR in the molding group was increased markedly, difference was statistical significance(P<0.01); Compared with the molding group,the value of ABR in the Chinese medicine group was decreased,differences were statistical significance(P<0.01).3.Cochlea basilar membrane stretched preparation in mice:Compared with the control group,cochlear hair cells of the molding group had disordered arrangement and serious deficiencies,and were smeared-out boundary.Compared with the molding group, cochlear hair cells in three Chinese medicine treatment group showed clear boundary,arranged line up and had little deficiencies,the higher the Chinese medicine,the better the cellular morphology.4.Serological changes in mice:Compared with the control group,the serum SOD activity decreased and MDA content improved,differences were statistical significance(P<0.01);PKC content improved in the molding group,difference was statistical significance(P<0.05).Compared with the molding group,in three Chinese medicine treatment group the serum SOD activity were gradually increased, differences were statistical significance(P<0.01);MDA content were gradually decreased, differences were statistical significance(P<0.01);and the PKC content were decreased,differences were statistical significance(P<0.05).5.Immunohistochemistry for PKC protein detection in mice : Compared with the control group,in the molding group the positive rate of PKC with mice cochlear were higher,differences were statistical significance(P<0.01).Compared with the molding group,in three Chinese medicine treatment group the positive rate of PKC with mice cochlear were lower, differences were statistical significance(P<0.01).6.Cochlear tissue SOD?PKC gene expression in mice:Compared with the control group,the gene expression of SOD with cochlear in the molding group were down-regulated and PKC gene were up-regulated,differences were statistical significance(P<0.01).Compared with the molding group,the gene expression of SOD with cochlear in three Chinese medicinetreatment group were up-regulated,and PKC gene were down-regulated,differences were statistical significance(P<0.01).Conclusion:Based on the free radical damage, the“Erlong Zuoci Pill” can increase the SOD activity ?decrease the MDA ?PKC content of model mice with presbycusis,achieving the therapeutic goal.