| Objective:Intermittent Hypoxia(IH)model was established to simulate the humanobstructive sleep apnea hypopnea syndrome OSAHS.The changes in the levels ofserum glucose,insulin content,insulin resistance,the levels of inflammatory markers,the tumor markers(TNF-a),interleukin-6(IL-6),interleukin-lbeta(IL-1?),in peripheral blood,and the changes in the expression level of c-Jun amino terminal kinase(JNK),p-JNK,nuclear factor kB(NF-kB)in the pancreatic tissue homogenate of rats were observed after Tempol antioxidant,to explore the intervention and inhibition of the antioxidants on the local inflammation andoxidative stress.Method:1.24 SPF adult male Wistar rats were selected to established the animal model,the weight was 200-220g.All the experimental rats were randomly divided into 4 groups,intermittent hypoxia group(group IH),anti oxidant intervention group(group IH+Tempol),normal antioxidant control group(group NC+Tempol)and normal control group(group NC),6 rats in each group were placed in 2 oxygen tanks and 3 of each oxygen chamber respectively.The daily intermittent hypoxia exposure time was 9Am-5Pm,and the total duration was 12 weeks.After thirteenth weeks,the rats in each treatment group were anesthetized,and then put to execution by bloodletting,portion of peripheral blood and pancreas tissues were reserved.2.After rats were exposed to intermittent hypoxia for 12 weeks.(1)Serum levels of glucose and insulin were measured,and the HOMA-IR of insulin resistance index was observed.(2)The levels of TNF-?,IL-6 and IL-1? in peripheral blood of each treatment group were detected by ELISA.(3)Pathological sections of pancreatic tissues from each group were observed to explore the pathological changes of pancreatic tissue after IH oxidation.(4)Western blot was used to detect the expression level of transcription factor JNK,p-JNK,cytoplasmic and nuclear NF-kB p65 in each group of pancreas.Results;1.The levels of fasting glucose and insulin and HOMA-IR in serum of each group:(1)Compared with the NC group,the glucose content in the serum of IH group and IH+tempol group increased obviously,the NC+tempol group had no obvious changes,and the IH+tempol group was lower than the IH group.There was a significant difference between the groups(F=8.235,P=0.000).The IH group was significantly different with the IH+tempol group,the NC+tempol group and the NC group(P<0.01),there was no difference in the rest groups.(2)Compared with the NC group,the serum insulin content in the IH and IH+tempol groups increased significantly,the NC+tempol group had no significant changes,and the IH+tempol group was lower than the IH group.There was a significant difference between the groups(F=66.672,P=0.000).Except there was no difference between the NC+tempol group and NC group,the other groups were significantly different(P value<0.01);(3)Compared with group NC,insulin resistance index in group IH and group IH+tempol increased significantly,NC+tempol group had no significant changes,and the IH+tempol group was lower than the IH group.There was a significant difference between group IH+tempol and IH group(F=76.172,P=0.000).Except there was no difference between the NC+tempol group and NC group,the other groups were significantly different(P value<0.01);2.Levels of TNF-?,IL-6 and IL-? related inflammatory markers in serum of each group.(1)Compared with group NC,the level of TNF-a in group IH and group IH+tempol was significantly higher,there was no significant change in NC+tempol group,and the level in group IH+tempol was lower than that in IH group.There was significant differencein each group(F=115.006,P=0.000),The IH group was significantly different with the IH+tempol group,the NC+tempol group and the NC group(P<0.01),there was no difference in the rest groups,(2)Compared with group NC,the level of IL-6 in group IH and group IH+tempol was significantly higher,the NC+tempol group had no obvious change,and the level in group IH+tempol was lower than that in IH group.There was significant difference in each group(F=39.988,P=0.000),Except there was no difference between the NC+tempol group and NC group,the other groups were significantly different(P value<0.05).(3)Compared with the NC group,the level of EL-1 beta in group IH and IH+tempol was significantly higher,the NC+tempol group had no significant change,the level in group IH+tempol was lower than that in IH group.There were significant differences in each group(F=27.919,P=0.000),Except NC+tempol group had no difference with IH+tempol group and NC group,the other groups were significantly different(P value<0.05).3.The pathological changes of pancreatic tissue:group IH compared with group NC,the lobule structure of the pancreas tissue was defected and even disappeared,the acinus tissue atrophied in varying degrees,the acinar cells arranged disorderly,the interval widened and the number of peripheral inflammatory cells increased obviously;The group of IH+tempol group was relatively complete and the acinus tissue was wilt compared with the IH group.The degree of shrinkage decreased,the number of peripheral inflammatory cells decreased,and there was a trend of normal recovery.4.The expression level of JNK,p-JNK,and NF-kB p65 after antioxidants intervention in pancreatic tissue.(1)Compared with group NC,the expression level of p-JNK in group IH and group IH+tempol was significantly higher,there was no significant difference in group NC+tempol,and the level in group IH+tempol was lower than that in IH group.There was a significant difference between groups(F=43.878,P=0.000).There was no significant difference in the relative expression of JNK protein between the groups(F=2.129,P=0.129).Compared with IH+tempol group,the relative expression of p-JNK protein in NC+tempol group and NC group was statistically significant(P<0.05),and there was no difference among other groups.(2)Compared with the NC group,the relative expression of NF-kB p65 in the cytoplasm of group IH and IH+tempol decreased obviously,the NC+tempol group had no obvious change,there was a significant difference in groups(F=50.176,P=0.000).Except there was no difference between the NC+tempol group and NC group,the other groups were significantly different(P value<0.01);(3)Compared with the NC group,the relative expression level of NF-kB p65 in the nucleus of group IH and IH+tempol increased obviously,the NC+tempol group had no obvious change.There was a significant difference in groups(F=67.551,P=0.000),Except there was no difference between the NC+tempol group and NC group,the other groups were significantly different(P value<0.01);Conclusions:1.Tempol antioxidants can decrease the level of glucose,insulin and insulin resistance in serum through intervention and inhibition of IH.2..Tempol antioxidant can reduce the level of inflammation induced by oxidative stress by IH,but it can not be completely reversed.3.Tempol antioxidants can reduce the infiltration of inflammatory cells around the injured pancreatic tissue through intervention and inhibition of IH,and there is a tendency to recover the lobular structure of the pancreas.4.The oxidative stress response induced by the intervention and inhibition of the oxidative stress induced by IH makes the decrease of the oxidative stress sensitive signal transduction pathway p-JNK and NF-kB p65 inflammatory signal transduction in the nucleus,the increase of the expression of NF-kB in the cytoplasm of the pancreatic tissue. |
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