| Objective:Ischemia-Reperfusion Injury(IRI)is an important pathology progress of clinical diseases,kidney as a hyperfused organ suffer from IRI easily,and lead to Acute Renal Failure.To explore the pathogensy and experimental therapy strategy, a lot of reasearches had been carried out ,but the pathogenesy of IRI is still obscure.Our research foucs on the espression of heat shock protein 70(HSP70)in renal Ischemia-Reperfusion Injury,to explore the regularity of HSP70 expression and its function in IRI,moreover, Activating Blood Circulation Herbs had been certified playing an important role in IRI.Our research is to reveal whether Activating Blood Circulation Herbs induce HSP70 synthesis and have a protective effect on IRI.Methods: 1. An experimental model of IRI using 108 rats with body weight about 200±20g were randomly divided into three groups ,group A: thirty-six rats ,operatied without ischemia.group B:thirty-six rats,clamp renal pedicle both side for 45 min inducing ischemia reperfusion at 0h,6h,12h,36h,48hpost-ischemia.venafemoralis injection NS.0.5ml/0.1Kg 5-10 min before operation.group C:thirty-six rats ,intervention with Activating Blood Circulation Herbs, clamp renal pedicle bothside for 45 min inducing ischemia reperfusion at 0h,6h,12h,36h,48h post-ischemia, vena femoralis injection Activating Blood Circulation Herbs 0.5ml/0.1Kg 5-10 min before operation; 2. annimal model establish:4%Chloral Hydrate 0.875ml?0.1kg peritoneal injection ,incisal opening from appendix ensiformis to articulation of pubis,exposed and obstructed renal pedicle for 45 min ,then opened bulldog clamp ,observed renal changing has been carried out.If successful occlusion,the color turned from darkred to black,while turn red quickly after reperfusion. 3. (1)biochemical examine Urea and CRE level at 10 min before operation and different time after post-reperfusion.(2)conserve one side renal sample in nitrogen canister at different reperfusion timing point for immunoblot. later.the other side renal sample paraffin imbed after retained in 10% Formaldehyde,for H-E stained and immunohistochemical .4. (1)detect group A,B,C Urea and CRE serum level at different time of reperfusion and before operation.(2)pathological observation.(3) to measure HSP 70 expression at different time by immunoblot and immunohistochemical.Result: 1. Renal function alter significantly between group B and A at 6h post-reperfision(p<0.05),show IRF has been develop after IRI. There is no significant difference between group A and B,but There is significant difference between group B and C (p< 0.05)show the interwention of Activating Blood Circulation Herbs have a positive effect on renal function destruction.2. Comparing with each group from pathological section ,we conclude that group C renal injury relieved after IRI ,which indicate that Activating Blood Circulation Herbs have a positive effect on renal function destruction.3. immunohistochemical results show that the expression of HSP70 significantly increased in cell nucleus of group C compared with group B .4. result of Western blot: the expression of HSP70 increase to accompany with different time of RIR in group C and B compared with group A ;the expression peak time of group B is at 24h after reperfusion ,while the peak time of group C is at 12h after reperfusion .The results indicate the Activating Blood Circulation Herbs have a positive effect on renal ischemia-reperfusion by inducing HSP70 expression increasing.Conclusion: 1. the control group express HSP 70 is low . 2. expression of HSP 70 increase after IRI and distribute in nucleus and cytoplasm. 3. expression of HSP70 increase significantly at 24h after IRI of group B while at 12h after IRI of group C. 4. Activating Blood Circulation Herbs have a positive effect on renal ischemia-reperfusion by inducing HSP70 expression increasing.All the conclusion may provide a proof for therapy strategy of renal ischemia reperfusion.
|
PDF Full Text Request