Effects Of Decoction Zuoci Pill On CAMP Content And PKA Expression In Cochlear Tissue Of Aged Deaf Mic

Purpose: To discuss the therapeutic mechanism of presbycusis by researching the effect of the Erlong Zuoci pill on c AMP content,PKA in tissue of mice with presbycusis.Material and method:Seventy five auricle responsive cleaning degree male Kunming mice were randomly selected fifteen as the blank control group,which were treated with NS in empty stomach.The other sixty were the presbycusis molding group,which were treated with D-galactose(150mg/kg)in empty stomach,once a day,with sixty days.After duplicating the mold,the presbycusis molding mice were randomly divided into the molding group,Erlong Zuoci pill low-dose group,mid-dose group and high-dose group.The blank control group and the molding group were drenched with NS,and the Erlong Zuoci pill low-dose group,mid-dose group and high-dose group were drenched with concentrated solution of Erlong Zuoci pill(0.3g/ml?0.6g/ml,1.2g/kg),once a day,with fourteen days.It observed morphological change of cochlear hair cells in mice to use cochlear basilar membrane whole-mount technique.It tested the c AMP content by using ELISA,the protein expression level of PKA to use IHC and the expression level of PKA and m RNA by using RT-PCR technique.Results: 1.To observe the general condition of mice Mice in the blank control group: Mice can normally feed and drink water with responsive and energetic,and thick and luster hair.After a long-term raise,the weight of them obviously increase and urine and stool are regular.Mice in model group: mice feed and drink less with serious aging phenomenon.Because of the obvious effect of D-galactose,mice appear the physical weakness and the weight loss,as more be static than moving.At the same time,mice show up the old state,such as withered and lack-luster hair with hair are sparse and loss,spine uplift,and poor mental state.Compared with the blank control group,mice in model group are easier to be captured.Mice in the low,medium and high dose groups of Erlong Zuoci pill: after the lavage treatment,the above situation of mice in each group equally appear different improvement compared with mice in the model group.Among them,mice in the high dose group improve obviously whose situation is closer to mice in the blank control group.2.observation on the basement membrane by cochlear morphology Mice in the blank control group: Mice are clearly can be seen the cells in cochlear inner and outer hair.The cells are the same size,distinct arrangement,clear boundary clear and uniform distribution.Cells in all layers are basically no lack.Compared with the cochlear pave piece of the mice in the blank control group,the cochlear pave piece of the mice in model group have the significant differences,with the internal and external hairy cells are untidy arrangement,disorder distribution,and different forms and sizes.Meanwhile they are serious lack or fall.Compared with the mice in the model group,the mice in Chinese medicine treatment group improved,with the internal and external hairy cells are arranged neatly,uniform distribution,and unobvious loss of hairy cells.There are significant differences between the mice in the high dose and in the model group.3.c AMP content in cochlear tissue ELISA detect results of c AMP content in cochlear tissue homogenate of the mice in each group: Compared with the mice in the blank control group,c AMP content in cochlear tissue homogenate of the mice in model group significantly reduced(P < 0.01).Compared with the mice in the model group,c AMP content of the medium and high dose of Erlong Zuoci pill obviously rise(P < 0.01).It shows that Erlong Zuoci pill have effective treatment with raising c AMP content in cochlear tissue.c AMP content of the low dose do not obviously increase.Compared with the model group,the statistics has no differences.4.PKA protein expression level in cochlear tissue Immunohistochemical detection results of the PKA protein expression level in cochlear tissue of the mice in each group: Compared with the mice in the blank control group,the PKA protein expression level in cochlear tissue of the mice in the model group decreased significantly(P < 0.01).Compared with the mice in the model group,the PKA protein expression level in cochlear tissue of the mice in the medium dose group increased(P < 0.05).the PKA protein expression level in cochlear tissue of the mice in the high dose group significantly increased(P < 0.01)It shows that the treatment of Erlong Zuoci pill can adjust the PKA protein expression level in the cochlear tissue mice,and the high dose takes the obvious effects.5.PKA m RNA expression level in cochlear tissue RT-PCR test results of PKA m RNA expression level in cochlear tissue of the mice in each group: Compared with the mice in the blank control group,PKA m RNA expression level in cochlear tissue of the mice in the model group decreased significantly(P < 0.01).Compared with the mice in the model group,PKA m RNA expression level in cochlear tissue of the mice in the medium dose group rise(P < 0.05),and PKA m RNA expression level of the mice in the high dose group significantly higher(P < 0.01).Conclusion: During the process of presbycusis,c AMP content and PKA expression level changes at the same time.By drug intervention of Erlong Zuoci pill,c AMP content and PKA expression levels in the cochlea tissue significantly elevated.It shows that Erlong Zuoci pill achieves the treatment of presbycusis maybe by adjusting c AMP content and PKA expression level in cochlear tissues.