Effects Of Treadmill Exercise Combined With IL-10 On The Recovery Of Nerve Function And Axonal Outgrowth In Rat Brain After Cerebral Ischemia

Objects To observe the effects of treadmill exercise combined with IL-10 on the recovery of nerve function and axonal outgrowth after cerebral ischemia and explore the related mechanisms of neural remodeling and functional reconstructionMethods 1. Construct lentiviral vector of IL-10 gene over expression and lentiviral vector of si RNA-IL-10. 2. Male adult Sprague-dawley rats were divided into 5 groups: sham-operated group(S group, n=12), MCAO group(M group, n=12) and MCAO + treadmill exercise group(M+E group, n = 12), MCAO + IL-10 gene over expression lentiviral vector group(M+V group, n = 12), MCAO + si RNA-IL-10 lentiviral vector + treadmill exercise group(M+Vsi+E group, n = 12). 3. MCAO model was established with the modified Zea-Longa’s method. 4. Rats in M+V and M+Vsi+E group were injected each with lentiviral vector of IL-10 gene over expression and lentiviral vector of si RNA-IL-10 through the lateral ventricle from the 12 hours after MCAO. 5. From the 48 hours after MCAO, rats in M+E and M+Vsi+E group were forced to run on the treadmill for 30 min per day for 2 consecutive weeks. Rats in other groups were left on the treadmill for 30 min and were not forced to run. 6. Rats in M group and M+E group underwent behavioral functional tests prior to the MCAO procedure in 1, 3, 7, 10 and 14 days after MCAO with modified neurologic severity scores(m NSS). 7.Use ELISA to detect expression of IL-10 around the ischemic brain tissue. 8. Immunohistochemistry was used to detect microglial cells activation specific protein IBA-1 expression around lateral ventricle.9. The tissue corresponding to the peri-infarct cortex was dissected for expression level of GAP-43, NF200 by Western blot, and detection NF200 by immunofluorescent staining.Results 1. Rats treated with the treadmill exercise had significantly lower m NSS scores at day 7(P<0.05)and 10,14(P<0.01) compared to the rats without exercise. 2. ELISA showed that the expression of IL-10 around the ischemic brain tissue higher than the rats without exercise(P<0.05). 3. Immunohistochemistry was found that microglial cells activation specific protein IBA-1 expression around lateral ventricle express more on the rats without exercise. 4. The results western blot and immunofluorescent showed that compared with M group, the expression of GAP-43 and NF200 higher on both M+E group(P<0.05, P<0.01) and M+V group(both P<0.01),while M+Vsi+E group lower than M+E group(P<0.01, P<0.05).Conclusions We demonstrates that treadmill exercise could inhibit the inflammatory reaction and recover of nerve function and promote axonal outgrowth in rat peri-ischemic brain area. That may be attributed, at least in part, to the regulation of IL-10.