The Effect And Mechanism Of Cyclosporin A Early Administration In Early Brain Injury In Rats Following Intracerebral Hemorrhage

objective:Cyclosporin A (CsA) Can be combined to CypD and hence inhibits mitochondrial permeability transition pore (mPTP) opening, maintaining mitochondrial homeostasis and Inhibiting Nuclear Translocation of Apoptosis-Inducing Factor(AIF), Thereby Inhibition of cell necrosis and apoptosis, Improve nerve function score. To assess the impact of early administration of cyclosporine A in rat experimental cerebral hemorrhage apoptosis and necrosis pathway.Methods:1.group of Experimental:randomly Assigned four equal groups of adult male SD rats, including the sham group(n=48), ICH+vehicle group(n=48), ICH+CsA5mg group(n=48), and ICH+CsAlOmg group(n=48).2.Model making and drug treatment:The rat ICH model was induced by injection of 0.1 ml autologous whole blood into the right basal ganglia(coordinates:0.2 mm anterior,3.5 mm lateral, and 5.5mm ventral to the bregma).Tail vein administration in 15min after surgery, More than 24h, once a day.3.neurological score: accroding to HuaYa professor evaluation method is adopted to improve the score, the greater the score on behalf of the lighter neural function damage, on the contrary, the heavier damage.4. Brain water content detection and brain edema:24h after cerebral hemorrhage with dry wet weight method to detect brain water content.24h with MRI detection of brain edema after cerebral hemorrhage.5.The blood-brain barrier detection:24h after cerebral hemorrhage with Evans blue method to detect the blood brain barrier permeability.6. Apoptosis detection:72h after cerebral hemorrhage with TUNEL method to detect apoptosis. 7.Cell necrosis detection:72h after cerebral hemorrhage with PI method to detect cell necrosis.8.Immune fluorescence detection:72h after cerebral hemorrhage with immune fluorescence detection CypD and AIF changes.9.Western blot detection:24h after cerebral hemorrhage with immune CypD imprint method, total protein and AIF AIF nucleoprotein. 10. Mitochondria to observe:72h electron microscope mitochondria morphology changes after cerebral hemorrhage.11.Cell morphology to observe:7d after cerebral hemorrhage with NISSL staining and HE staining to detect cell morphological structure.12.statistic analysis:Use SSPS 17.0 software, the results are to Mean and SD, said using single factor analysis of variance, between group compared with the student t test analysis, P< 0.05 for significant standard.Results:1.72h of neural function after cerebral hemorrhage score significantly difference in the Sham group (p<0.01), CsA treatment group was significantly more ICH group (p<0.01), for example, CsA+Vehicle group (p<0.01).2.24h after cerebral hemorrhage brain water content significantly increased in the Sham group (p<0.01), for example+CsA5mg, ICH+CsA10mg group decreased significantly in the ICH group (p<0.01).24h MRI scan line shown cerebral edema after cerebral hemorrhage significantly increase, a significant reduction in treatment group.3.24h after cerebral hemorrhage blood brain barrier permeability significantly damage than the control group (p<0.01), for example+CsA5mg, ICH+CsAlOmg group compared with, for example (p<0.01).4. Cerebral hemorrhage after 72h of apoptotic cells was significantly more than in the Sham group (p<0.01), for example+CsA5mg group, ICH+CsA10mg apoptotic cells was significantly reduced in the ICH group (p<0.01), for example the ICH+CsA5mg group CsAlOmg group of apoptotic cells was significantly increased (p<0.01).5. Cerebral hemorrhage after 72h of necrotic cells was significantly more than in the Sham group (p<0.01), for example+CsA5mg group, ICH+CsA10mg necrotic cells was significantly reduced in the ICH group (p<0.01), for example the ICH+CsA5mg group CsAlOmg group of necrotic cells was significantly increased (p<0.01).6. 24h after cerebral hemorrhage with immune CypD and AIF imprint method, the change of the total protein and AIF nucleoprotein express significantly enhanced in the Sham group (p<0.01), for example+CsA5mg, ICH+CsAlOmg group decreased significantly in the ICH group (p<0.01).7.72h mitochondria edema after cerebral hemorrhage, ICH+CsA5mg group, ICH+CsA10mg mitochondria edema improved.8. Cerebral hemorrhage after 7d cells attenuation, ICH+CsA5mg group, ICH+CsAlOmg change cell morphology and cell number did not see obvious decay.Conclusion:In ICH model, CsA could Reduce the brain water cont ent and alleviate Early cerebral edema, to improve the early blood brain barrier permeability. Cyclosporin A Can be combined to CypD and hence inhibits mitochondrial permeability transition pore opening, Thereby maintaining mitochondrial homeostasis and inhib iting Nuclear Translocation of apoptosis Inducing Factor, Inhibition of cell apoptosis and necrosis, Improve early brain injury, Improve the neurological function score.