| Objective:1.To investigate the injury effect of diabetes on cerebral ischemia reperfusion model.2.To investigate the neuroprotective effects of GLP-1 /GIP double receptor agonist DA3-CH on cerebral ischemia reperfusion injury model and associated diabetes model(behavioral behavior,infarction area,inflammatory factors).3.To investigate the effect of DA3-CH on the inflammatory response of diabetic rats after cerebral ischemia-reperfusion injury model and the possible mechanism of neuroprotection.Methods: fifty four male SD rats were randomly divided into five groups:(1)control group(con,n=6),(2)diabetic cerebral ischemia-reperfusion group(DM+I/R,n=12),(3)cerebral ischemia-reperfusion group(I/R,n=12),(4)cerebral ischemia-reperfusion group(DA3+I/R,n=12),(5)diabetic cerebral ischemia-reperfusion group(DM+DA3+I/R,n=12).The diabetic model was made by STZ.The middle cerebral artery occlusion model was made by thread bolt method.The thread bolt was removed for reperfusion after 2 hours of ischemia.The DA3-CH groups were injected intraperitoneally with DA3-CH injection(10nmol/kg/d)14 days before ischemia-reperfusion,and the cerebral ischemia-reperfusion group was injected intraperitoneally with the same amount of normal saline at the same time.Blood glucose and body weight were measured at regular intervals,and nerve defect score(longa score)was performed 24 hours after ischemia-reperfusion.The ratio of infarct volume was detected bytcl-2,3,5-triphenyltetrazolium(TTC)staining.The expression of IL-1?,TNF-?,NF-? B in ischemic penumbra was detected by Western blot?Results:1.Comparison of blood glucose in each group: blood glucose in each diabetes model group(DM+I/R,DM+DA3+I/R)increased significantly before and after diabetes modeling(4.97±0.4 VS 22.82±7.04;5.10±0.4 VS 25.37±0.7);Compared with the control group(con)and the control group(I/R),blood glucose in the diabetes model group(DM+I/R)was significantly increased(22.82±7.04 VS 5.58±1.09;22.82±7.04 VS 5.29 ± 0.78),the difference was statistically significant(p<0.001).There was no significant difference in blood glucose between the DA3+I/R group and the I/R group(5.40 ± 0.88 VS 5.29 ± 0.78)(P>0.05).There was no significant difference in blood glucose between DM+DA3+I/R group and DM+I/R group(25.37±0.7 VS 22.82±7.04)(P>0.05).2.Weight comparison of each group: before and after the diabetes model group(DM+I/R,DM+DA3+I/R),the weight of each group decreased significantly(291.7 ±10.5 VS 277± 7;295 ± 11.14 VS 292.7± 10.44);Weight loss in the diabetes model group(DM+I/R)was(292.6±10.12 VS 303.6±10.67)compared with that in the control group(con)and the cerebral ischemia-reperfusion model group(I/R).292.6±10.12 VS301.2 ± 12.81),the difference was statistically significant(p<0.05).There was no significant difference in body weight between the DA3+I/R group and the I/R group(298.4±5.76 VS 301.2±12.81)(P>0.05).There was no significant difference in body weight between DM+DA3+I/R group and DM+I/R group(292.7±10.44 VS 277±7)(P>0.05).3.Comparison of neurological impairment scores in each group: the DM+I/R group was compared with the I/R group(2.39 ± 0.50 VS 1.67 ± 0.69),and the degree of neurological impairment was aggravated(p < 0.001).Compared with the DM+DA3+I/Rgroup(1.78±0.64 VS 2.39±0.50),the function of nerve defect in DM+DA3+I/R group was significantly improved(p<0.05).Compared with the I/R group,DA3+I/R group(1.78± 0.64 VS 1.67 ± 0.69)showed improvement in neurological impairment,but no statistical difference(p>0.05).4.Comparison of cerebral infarction volume in each group: compared with the DM+I/R group and the I/R group,the cerebral infarction volume ratio increased significantly,with statistically significant difference(p<0.001).Compared with DM+DA3+I/R group,the cerebral infarction volume ratio decreased significantly(p<0.001).The cerebral infarction volume ratio of DM+DA3+I/R group was also significantly lower than that of DM+I/R group(p<0.05).5.Western Blot results The expression levels of TNF-?,IL-1?and NF-?B in the midbrain tissues of each group were detected.The results were as follows:5.1 comparison of TNF-?expression levels in the DM+I/R group and the I/R group,TNF-?expression levels in the DM+I/R group increased(p<0.001).TNF-?expression level decreased in DM+DA3+I/R group compared with DM+I/R group(p<0.001).TNFexpression in DA3+I/R group decreased compared with that in I/R group(p<0.05).TNFexpression level was also significantly decreased in DM+DA3+I/R group compared with I/R group(p<0.05).5.2 comparison of IL-1? expression levels in each group: IL-1? expression level in DM+I/R group was significantly increased compared with that in I/R group(p<0.001).The expression level of IL-1 ? in DM+DA3+I/R group was significantly reduced compared with that in DM+I/R group(p<0.001).The expression level of IL-1?in DA3+I/R group was decreased compared with that in I/R group(p<0.05).IL-1 ?expression was decreased in DM+DA3+I/R group compared with I/R group(p<0.05).5.3 comparison of NF-? B expression levels in each group: compared with the DM+I/R group,NF-?B expression levels in the DM+I/R group increased significantly(p<0.001).NF-? B expression decreased in DM+DA3+I/R group compared with DM+I/R group(p<0.001).NF-?B expression decreased in the DA3+I/R group compared with the I/R group(p<0.05).NF-? B expression increased in DM+DA3+I/R group compared with I/R group(p<0.05).Conclusion:1.It was confirmed that diabetes aggravates cerebral ischemia-reperfusion injury by increasing the infarction area,aggravating the neurological deficit score and promoting the expression of inflammatory factors(TNF-??IL-1??NF-?B),and inflammation plays an important role in aggravating cerebral ischemia-reperfusion injury caused by diabetes.2.DA3-CH has a neuroprotective effect on cerebral ischemia-reperfusion injury model.3.DA3-CH has a significant protective effect on the model of diabetic patients with cerebral ischemia-reperfusion injury,and its mechanism may be related to the alleviation of neuroinflammation. |
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