Based On JNK Pathway To Explore The Intervention And Mechanism Of Ligustrazine Restricting The Occurrence Of Ototoxicity And Nephrotoxicity Caused By Gentamicin

ObjectiveBased on the functionality of JNK pathway played in the occurrence of ototoxicity and nephrotoxicity caused by gentamicin,explore the intervention mechanism of ligustrazine.Further elaborate relationship between ear and kidney in the kidney main ear theory.Methods48 healthy and hearing responsive white red-eye guinea pigs were screened;then randomly divided them into four groups named gentamicin(GM)group,ligustrazine(TMP)group and(GM+TMP)group and normal blank(NB)group respectively.Each group has twelve guinea pigs.In the GM group all guinea pigs did GM intramuscular injection(IM)based on the amount of 120mg·kg-1·d-1;in the GM+TMP group,all guinea pigs did GM IM and TMP IM based on the amount of 120mg·kg-1·d-1 and 30mg·kg-1·d-1;in the TMP group all guinea pigs did TMP IM based on the amount of 30mg·kg-1·d-1 respectively;in the NB group,all guinea pigs 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.After that all all guinea pig were accept the blood urea nitrogen(BUN)and creatinine(SCr)level test.Once the final test was finished,all guinea pigs’cochleas,kidneys,livers were prepared to be formalin-fixed paraffin-embedded format.HE dye were done and the structure of these organization were observed under the light microscopy.Western blot method was used to examine cochleas and kidneys’JNK level.Finally,correlation analysis was used.Results1.From the results got from click ABR test,the hearing threshold(HT)of all guinea pigs from each group has no significant differences before IM.(P>0.05).After IM,compared with the HT got form NB group(8.17±5.33 dB nHL),the HT got from the GM group(54.64±23.72 dB nHL)and GM+TMP group(28.17±26.54 dB nHL)increased significantly(P<0.01).Though the HT of GM+TMP group also had increased,but it was littler than that of GM group.There was no significant difference between TMP group(7.83±4.49 dB nHL)(P>0.05).2.From the results got from renal function test,the SCr and BUN of GM group were significantly increased after IM,and compared with the other three groups,there were differences between of them(P<0.05).The SCr and BUN of GM+TMP group、TMP group and the NB group also had difference between of each other,but there were no difference(p>0.05).3.From the results got form morphology,the morphology of cochlea got from NB 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 NB group,occasionally a small amount of protein tube wil be seen.From the observation of liver,there were no significantly differences among four groups.4.From the results got form western blot,the JNK2 expression level of GM group were increased after IM,and there were significantly differences with other three groups(P<0.05).The JNK2 expression level of GM+TMP group were also increased after IM and there were significantly differences with TMP and NB groups(P<0.05),but it was littler than that of GM group(P<0.05).The JNK1 expression level were no significantly differences among four groups.Conclusion:1.Deafnes and renal function impairment model was established by injected muscle gentamicin.JNK pathway plays a regulary role in the ototoxicity and nephrotoxicity of gentamicin.JNK pathway was activated and JNK protein were expressed.2.TMP can prevent toxicological effects of gentamicin,then protect the function of hearing and renal.JNK pathway plays a regulation role in restricting the occurrence of ototoxicity and nephrotoxicity caused by gentamicin.JNK pathway was suppressed and decreased JNK expression level3.The correlation analysis showed the kidney had the same occurrence of pathology and prevention with ear.JNK may be one of the indicators between ear and kidney,especially JNK2,that may be one of the molecular basis of kidney main ear theory.