Protective Effects And Mechanism Of Salvla Miliorrhiza Bge.f.alba C.y.wu Et H.w.li To Rat Following Focal Cerebral Infarction

Objectives:To explor the dynamical progression of pathology and the protective effects of Salvia miltiorrhiza Bge.f.alba C.Y.Wu et H.W.Li in rats following focal cerebral infarction.Methods:To establish the model of focal cerebral infarction based on the principle of photochemical initiation of thrombosis. To measure regional cerebral blood flow (rCBF) in the ischemic core and penumbra by Laser Doppler Flowmeter. The area of the red or blue field in cerebral cortex was calculated with the system of image-proplus4.0. The numbers and anatomic distribution of necrosis or apoptosis cells were observed in situ histologically with the use of hematoxylin-eosin (H&E) staining and terminal deoxynucleotide tranferase (TdT)-mediated dUTP nick end-labeling (TUNEL) assay in adjacent sections. c-Fos, HSP70 and Caspase-3 positive cells were observed by immunohistochemical method at different time points.Results:1 rCBF in ischemic core looked like waves. It decreased to the minimum after 3 h in which it was (9.36±1.72)% of normal Cerebral blood flow. But after 6 h it became to increase. rCBF increased to the maximum at 12 h in which it was (31.18±1.44)% of normal flow. It reduced after 24 h. At 24,36 and 48 h points, rCBF was no significant difference than 3 h. rCBF in penumbra decreased to the minimum at 6 h in which it was (30.88±7.11)% of normal flow. From the points of 9 to 48 h, rCBF in penumbra increased gradually. At 48 h (91.90±4.56)% of normal flow was recovered which was significant difference than others.2 The red field occurred at 30 min following focal cerebral infarction and expanded with time. The peak occurred between 48 h and 3 d. At 4,5 and 6 d, the red field decreased slightly which were no significant differences than it of 3d. The area of the blue field displayed doublepeak. There was an increasingly tendency from 3 h. At 9 h the first peak occurred. After 12 h it began to decrease. The second peak occurred at 24 h and it was more higher than the first (P<0.05). After that it decreased and disappeared.3 Necrosis cells were predominantly focused in the infarction. While apoptosis cells were perferentially located in the milieu of infarction. Both necrosis and apoptosis cells were all expending in a radial fashion. Necrosis cells appeared in the core of lighing at 3 h following focal cerebral infarction. But apoptosis cells only occurred at 6 h. The numbers of necrosis or apoptosis cells were progressively increased with time. The peak of necrosis cells occurred at 24 h and apoptosis cells'occurred at 3 d.4 c-Fos positive cells expressed generously at the point 30 min. The expression was increasing with time. The high peak of expression occurred at 6 h and all two cerebral hemisphere expressed. But at 30 min the expression was only in the injured hemisphere. From the time point of 9 h, the expression of c-Fos positive cells decreased gradually and the expression occurred only in the milieu of infarction. At all time points there were no expression in the infarction.5 HSP70 was expressed in the milieu of infarction at 12 h. At 48 h, HSP70-immunoreactivities were increased to the maximum in the milieu of infarction. Immunohistochemical analysis revealed no HSP7-immunoreactivity in the contral and sham operative groups.6 Caspase-3 was expressed in the milieu of infarction at 30 min. Caspase-3-immunoreactivities were increased to the maximum in the milieu of infarction between 36 and 48 h. However, there were no Caspase-3-immunoreactivity in the contral and sham operative groups.7 The area of red and blue field were more small in the group of Salvia miltiorrhiza Bge.f.alba C-Y-Wu et H-W-Li than the simple ischemia group. The dynamical progression in expression of HSP70 and Caspase-3 at different time points in the Group of Salvia miltiorrhiza Bge.f.alba G·Y·Wu et H·W·Li were in accord with in the group of simple ischemia. But the number of HSP70-immunoreactivities in the group of Salvia miltiorrhiza Bge.f.alba C·Y·Wu et H·W·Li was more intense than in the Group of simple ischemia. Caspase-3-immunoreactivities in the Group of Salvia miltiorrhiza Bge.f.alba C·Y·Wu et H·W·Li was more rare than in the Group of simple ischemia. There were significant differences between the group of low and middle, low and high dose (P<0.05). But there were no significant differences between the group of middle and high dose (P>0.05).Conclusions:1 rCBF in the infarction and penumbra following focal cerebral infarction showed significant rules in time and space. The recovery of rCBF in penumbra, the doublepeak of the area of blue field and the progressive expanding of infarction showed that the model could induce ischemia/reperfusion injury which triggered delayed neurocyte apoptosis.2 Necrosis cells were predominantly focused in the infarction. While apoptosis cells were perferentially located in the milieu of infarction. Both necrosis or apoptosis cells were expending in a radial fashion. Anti-apoptosis may be one of significant methods in treating cerebral infarction because delayed apoptosis.3 Our study demonstrated that focal cerebral infarction could induce neurons apoptosis and the expression of HSP70, Caspase-3.The expression of HSP70, Caspase-3 and neurons apoptosis all occurred in the farction's milieu. The peaks of HSP70 and Caspase-3 were earlier than cell apoptosis's. Caspase-3 mediated delayed neurocyte apoptosis. c-Fos could be induced by focal cerebral infarction,but there were remote expression which were no distinct mechanism.4 The mechanism of the neuroprotective effects might be that Salvia miltiorrhiza Bge.f.alba C·Y·-Wu et H·W·Li could decrease neurons necrosis and apoptosis through increasing the expression of HSP70 and decreasing the expression of Caspase-3.