The Effect Of Clausenamide On The Expression Of Bcl-2 And Apoptosis After MCAo And Reperfusion In Renovascular Hypertensive Rats

Objective To study the effect and mechanism of clausenamide on the expression of Bcl-2 and apoptosis after MCAo and reperfusion in renovascular hypertensive rats. Methods: (1) Double renal artery stenosis of the outbred health male Wistar rats aged 1.5 months were performed with silver clips with 20-mm inner diatemer, After this operation,when the rats blood pressure were high,they were refered as to renovascular hypertensive rats. (2) The one hundred rats weighting 290~310g without stroke were randomly divided into four groups(25 each group): clausenamide 30mg intervenient group, clausenamide 15mg intervenient group, single ischemia and reperfusion model group and sham-operated group. The middle cerebral artery occlusion (MCAo) was made by intraluminal vascular suture for 2h and reperfusion in the rats, but MCAo was not performed in sham-operated group. Computerized pathological image analyzer was used to measure the cell counts of bcl-2 with Immunohistochemical staining and the counts of apoptosis with TUNEL staining methods respectively in coronal sections of brain after reperfusion (6, 12, 24, 48 and 72 hours). Results: (1) The Bcl-2 protein had expressed 6h after reperfusion, andreached its height at 24h, then declined gradually. The Bcl-2 protein positive cell counts at every times in clausenamide intervenient groups were significant higher than another group (p<0.01), and the counts was a significant high in clausenamide 30mg intervenient group than that in clausenamide 15mg intervenient group (p<0.05). (2) The number of apoptotic cells increased with the development of reperfudion, and reached its height at 72h. The apoptosis counts of every times in clausenamide 30mg intervenient group was significant lower than another two groups (p<0.01).But, there was no significant difference between clausenamide 15mg intervenient group and single ischemia and reperfusion model group 48h after reperfusion (p>0.05). There was remarkable low at every times in clausenamide 30mg intervenient group than that in clausenamide 15mg intervenient group(p<0.05). No positive cells of Bcl-2 protein and apoptotic cell could be discovered in brain section from sham-operated animals and in the oppose side to ischemia from the other groups animals.Conclusion: Expression of Bcl-2 protein increases and apoptosis exists after focal cerebral ischemia and reperfusion in rats' brain. Clausenamide can enhance the expression of Bcl-2 protein and inhibit remarkably apoptosis, which have a relationship between dose and reaction. Clausenamide may cooperate with Bcl-2 in inhibiting apoptosis. that may be the mechanisms of clausenamid to protect brain cells from ischemic cell damage.