The Role Of AIF In Ligustrazine Resistance To The Gentamicin-induced Oto-renal Injury In Rat Model

ObjectiveBased on the functionality of AIF pathway played in the occurrence of oto-renal Injury caused by gentamicin,explore the intervention mechanism of ligustrazine and provide a supplementary method to the treatment of ototoxicity and nephrotoxicity.It elaborates the pathological correlation between ear and renal,which enrichs kidney main ear theory.MethodsInitially,40 healthy and hearing responsive white SD rats were screened;then randomly divided them into four groups named normal saline(NS)group,gentamicin(GM)group,ligustrazine(TMP)group and GM+TMP group respectively.In the GM group all SD rats did GM intramuscular injection(IM)based on the amount of 120mg·kg-1·d-1;in the TMP group all SD rats did TMP IM based on the amount of 30mg·kg-1·d-1;in the GM+TMP group,all SD rats did GM IM and TMP IM based on the amounnt of 120mg·kg-1·d-1 and 30mg·kg-1·d-1 respectively;in the NS group,all SD rats did NS IM using the same amount as the GM group did.All IM required being last for 13 days.Before IM and the first day after IM,click ABR were tested.Once the final test was finished,all SD rats'cochleas and kidney specimens were prepared to be formalin-fixed paraffin-embedded format.After that cochlear slices and kidney specimens HE dye were done and the structure of Corti's organ and inner cell in the spiral ganglion were observed.Western blot method was used to examine AIF expression level.Results? From the results got from click ABR test,the hearing threshold(HT)of all SD rats from each group has no significant differences before IM(P>0.05).After IM,compared with the HT got form NS group,the HT got from the GM group and GM+TMP group increased significantly(P<0.01);there was no significant difference between NS group and TMP group(P>0.05);compared with the NS group,although the HT got from GM+TMP and GM group increased,there were no significant differences(P>0.05)between both groups with the HT got from GM+TMP group increased less.?The CREA and Bun of GM group were significantly increased,and compared with the other three groups,there were differences between of them(P<0.05).Among of tnem,though GM+TMP group also had increased,but it was littler than that of GM group;After IM,GM+TMP group?TMP group and the control group also had certain difference between of each other,but there were no difference(p>0.05).?From the observation of cochlear slices,the histomorphology of cochlea got from NS group was normal;the Corti's organ got from GM group was severely deformative with OHC swelling and deformation and lots of cells missing and cavitation in the spiral ganglion;however,from the GM+TMP group,the situation was slightly better;in the TMP group,the structure of Corti's organ and cells in the spiral ganglion were normal but with some slightly changed.From the observation of kidney,the renal tubulars were holonomic,and tubules were arranged closely;the renal tubulars epithelial cells of the GM group have dropsy,and renal tubulars expanded,we can see protein tube type and cell tube type;GM+TMP group is better than the model group,occasionally a small amount of protein tube wil be seen.? From the western blot results,after one-way analys is of variance and comparison between groups,for cytoplasm protein,it can be seen the density of AIF of cochlear in different groups were NS group>GM+TMP group?TMP group>GM group;The density of AIF in GM group changed much,compared with the other three groups,there were differences between of them(P<0.05).TMP group is amost the same of that of GM+TMP group.What's more,they all littler than that of normal group.But there were no difference between of the there groups(p>0.05).The density of AIF in kidney in different groups were NS group>TMP group>GM+TMP group>GM group.Compared with the normal group,GM group and GM+TMP group were all changede much,there were difference(p<0.05).Though TMP group had changed a little,it was almost the same of that of NS group.The density of AIF for GM+TMP group was still very high,and there was no difference with GM group.There was no difference between GM+TMP group and TMP group(p>0.05).For caryon protein,it can be seen the density of AIF of cochlear in different groups were GM group>GM+TMP group>NS group?TMP group;The density of AIF in GM group changed much,compared with the other three groups,TMP group and NS group have differences.TMP group is amost the same of that of NS group.What's more,GM+TMP group is littler than that of GM group.But there were no difference between of them(p>0.05).For kidney,no matter the cytoplasm protein or the caryon protein,the density of AIF is all the most same.Conclusion:? GM is ototoxic and nephrotoxic,and it can lead to the abnormal of the histomorphology of cochlear and kidney.The role of AIF signal pathway can regulate and control tne oto-renal injury induced by GM.? TMP can improve HT,protect kindey,decrease AIF expression level from molecular aspects,inhibit the phosphorylation of AIF and may effect AIF pathway and play a treatment role in ototoxic deafness.