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Objective To observe the effect ofmitochondrial division inhibitor (Mdivi-1) on the neuronal apoptosis via mitochondria signaling pathways in rats after cerebral ischemia/reperfusion (I/R) and its possible mechanism.Methods Sixty male adult healthy Wistar rats were chosen and randomly divided into sham-operated group,vehicle group and Mdivi-1 pre-conditioning group (n=20); animal models of middle cerebral artery occlusion/reperfusion in the later 2 groups were established by a filament method in the left extemal-internal carotid artery.Rats were injected with 1.2 mg/kg Mdivi-1 in the Mdivi-1 pre-conditioning group and dimethyl sulfoxide in the vehicle group 15 minutes before the cerebral ischemia.After 24 h of reperfusion,apoptotic cell death was assessed by TUNEL staining,the cytochrome C expression (Cyt C) was detected by SABC immunohistochemical staining,the protein level of Cyt C in the cerebral issues was detected by Western blotting and the mRNA expression was measured by semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR).Results The apoptosis rate (45.49％±0.77％),Cyt C positive cell rate (55.12％±1.47％) and protein and mRNA expressions of Cyt C (0.932 ±0.001,0.789±0.009) in the vehicle group were significantly higher that in the sham-operated group (4.16％±0.25％,4.22％±0.20％,0.395±0.002,0.467±0.003,P＜0.05).The apoptosis rate (45.49％±0.77％),Cyt C positive cell rate (55.12％±1.47％) and protein and mRNA expressions of Cyt C (0.594±0.002,0.523±0.004) in the Mdivi-1 pre-conditioning group were significantly decreased as compared with those in the vehicle group (P＜0.05).Conclusion Mitochondrial division inhibitor Mdivi-1 might reduce neuronal apoptosis by inhibiting mitochondria cytochrome C apoptotic pathway to protect cerebral against ischemia/ reperfusion injury.
RESUMEN
Objective To explore the effect of SA liposome mediated human interleukin-10 (IL-10) gent transfection on NHE-1 mRNA expression in penumbra area following focal cerebral ischemia reperfusion injury in rats. Methods Totally 78 male SD rats were randomly divided into normal control group (n=6), MCAO group (n=24), hIL-10 transfection group (n=24) and empty vector transfection group (n=24). Longa's method was employed to establish MCAO models in the latter 3 groups. The rats in the MCAO group underwent stereotactic operation without drug injection, and the hIL-10 transfection group and empty vector transfection group were injected stereotactieally with pcDNA3.1-IL-10 and pcDNA3.1, respectively, both by SA liposome mediation. After transfection, RT-PCR and ELISA were used to determine the effect of transfection, TTC staining was conducted to detect the infarct volume. Meanwhile, real-time quantitative PCR was performed to examine the expressions of NHE-1 mRNA and NF-κB mRNA in the penumbra area. Results (1) SA liposome effectively mediated the hIL-10 gene to transfect the brain tissue. Also hIL-10 gene transfection played neuroprotective effect by reducing the brain infarct volume. (2) The expression of NF-κB mRNA in different groups was 1.00±0.33, 4.76±0.41, 4.58±0.62 and 2.77±0.43, respectively, hIL-10 gene transfection also inhibited the increase of NF-κB mRNA expression in the penumbra area following the cerebral ischemia reperfusion injury. (3) The expression of NHE-1mRNA was 1.00±0.22, 4.16±0.48, 3.97±0.51 and 2.82±0.47, respectively, hIL-10 gene transfection also inhibited the increase of NHE-1 mRNA expression in the penumbra area following the cerebral ischemia reperfusion injury. Conclusions The hIL-10 transfection can exert the protective effect on the brain against cerebral ischemia-reperfusion injury partly via inhibiting the NHE-1 mRNA expression.