| Purpose:The effect and mechanisms of Shenqi pill to Notch2/hesl Signaling Pathway in gentamicin-induced the rat-oto-renal injury modelMethods:The mature SD rats were randomly divided into normal group, model group, DAPT group, Shenqi pill group. We use Gentamicin model ing (120mg/(kg·d), intraperitoneal injection, continuous ten days). Before modeling and the1st,14th,28th day post-modeling,to detect hearing changes by auditory brainstem response (ABR); To observe the cochlea hair cell damage and repair with cochlea stretched. To observe the damage situation of renal tubules with light microscope and electron microscopy. To observe the percentage of Ki67-positive cells in oto-renal by immunofluorescence. Immunohistochemistry observed the expression of Jag2/Notch2/hesl signal in the tubular. By western-blotting and real-time quantitative PCR, observed expression of the Jag2/Notch2/hesl in cochlea and in kidney tissue in the1st,14th,28th post-modeling.Results:Renal biopsy showed:In the normal group, the tubular no significant change in each time. In Model group, injury of the renal tubular is severe in the1st post-modeling; it is mitigation in the14th; the damage of tubular return to normal in the28th; in the1st14th, compared with the model group, the degree of kidney damage is obvious mitigation in the DAPT group; in the28th, tubular back to normal in DAPT. In the1st14th, compared with the model group, the degree of kidney damage is obvious mitigation in the Shenqi pill group; in the28th, tubular back to normal in the Shenqi pill. Immunohistochemistry, Western blot protein, real-time quantitative PCR show that the model group compared with the normal group, the level of Jag2/Notch2/hes1is significantly higher in the1st,14th (P<0.05); there is no significant difference in the28th (P>0.05); compared with the the model group, the level of Jag2/Notch2/hes1is significantly decrease in DAPT group (P<0.05); in the28th there is no significant difference between the model group and DAPT group (P>0.05). Compared with1st,14th, the Jag2/Notch2/hes1expression decreased (P<0.05) in the DAPT group in the28th. In the1st,14th, compared with the model group, the level of Jag2/Notch2/hes1is significantly decrease in the Shenqi pill group (P<0.05); in the28th, there is no significant difference between the model group and the Shenqi pill group (P>0.05); Compared with1st,14th, the Jag2/Notch2/hes1expression decreased (P<0.05) in the Shenqi pill group in the28th. Immunof luorescence showed:the percentage of Ki67-positive cells was2.52%in the normal group; Compared with the the normal group, the percentage of Ki67-positive cells was significant increased in the model group in the1st,14th. Compared with the the model group, the percentage of Ki67-positive cells was significant increased in the DAPT group and the Shenqi pill group in the1st,14th.Auditory brainstem response (ABR) shows:the normal group had no significant change; compared with before treatment, the ABR threshold were significantly higher in model and DAPT and Shenqi group. Compared with model group, the ABR threshold is a significantly downward in the DAPT group. Compared with the model group, the ABR threshold were significantly decreased in Shenqi pill group. Cochlea shtreched show:damage of hair cell is severe in cochlea in the1st14th,28th after modeling in gentamicin group; and compared with the1st, hair cell damage is significantly reduced in the28th. Compared with the model group, damage of hair cell is reduced in the1st,14th,28th after modeling in DAPT group. Compared with the1st, hair cell damage is significantly reduced in the28th in the DAPT group. Compared with the model group, damage of hair cell is reduced in the1st14th,28th after modeling in the Shenqi pill group; and compared with the1st, hair cell damage is significantly reduced in the28th in the Shenqi pill group. Real-time quantitative PCR and western-blotting shows:compared with the normal group, the expression level of Jag2/Notch2/hesl is significantly higher in the1st,14th,28th in the model group (P<0.05); compared with the1st, the level of Jag2/Notch2/hesl is significantly downward in the28th in the model group (P<0.05). At the same time period compared with the model group, the expression level of Jag2/Notch2/hesl is significantly lower in DAPT group (P <0.05); compared with the1st, the expression level of Jag2/Notch2/hesl had evident decline in the28th in the DAPT group (P<0.05). At the same time period compared with the model group, the expression level of Jag2/Notch2/hesl is significantly lower in the Shenqi pill group (P<0.05); compared with the1st, the expression level of Jag2/Notch2/hes1had evident decline in the28th in the Shenqi pill group (P <0.05). Immunofluorescence showed:The percentage of Ki67-positive cells was1.42%in the normal group; Compared with the the normal group, the percentage of Ki67-positive cells was significant increased in the model group in the1st,14th,28th. Compared with the the model group, the percentage of Ki67-positive cells was significant increased in the DAPT group and the Shenqi pill group in the1st,14th,28th.Conclusion:Jag2/Notch2/hes1signaling pathways may be one of the important role in the gentamicin-induced renal and ear injury and repair process. Shenqi pill and DAPT may be go through inhibit Jag2/Notch2/hes1signaling pathways to reduce ear and renal injury and promote tubular epithelial cell and the hair cells repair. |
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