Protective effects of Breviscapine injection against focal cerebral ischemia-reperfusion injury in rats
- VernacularTitle:灯盏花素注射液对大鼠脑缺血再灌注局灶性脑损伤的保护作用
- Author:
Xingsan TANG
;
Yazhen MA
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2005;9(29):243-245
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: Breviscapine, extracted from a Chinese herb Erigeron breviscapus (Vant.) Hand-Mazz (Bre), has remarkable activities against platelets and thrombus formation. It can protect against ischemic brain damage through eliminating free radicals and apoptosis.OBJECTIVE: To study the protective effects of Breviscapine on brain damage in rats after ischemia-reperfusion injury, taking MgSO4 as control.DESIGN: Randomized controlled study and analysis of variance.SETTING: College of Life Science and Technology, Xiaogan University.MATERIALS: This experiment was conducted in the College of Life Science and Technology of Xiaogan University between May 2004 and November 2004. Forty male Wistar rats were selected and randomized into five groups: sham-operation group, cerebral ischemia-reperfusion (IR)group, MgSO4 group, 50 mg/kg Breviscapine group and 75 mg/kg Breviscapine group with 8 in each group.METHODS: Rats were subjected to cerebral ischemia-reperfusion induced by inserting a nylon thread into the internal carotid artery to block the origin of middle cerebral artery and removing the thread later. Normal saline of 20 mL/kg was administrated in sham-operation group and IR group, while 50 mg/kg, 75 mg/kg Breviscapine and 30 mg/kg MgSO4 were administrated in other groups respectively 10 minutes after the onset of ischemia. Neurological scoring was performed 1 hour, 2 hours, 5 hours and 23 hours after cerebral ischemia-reperfusion (Five points in total: 0 as unobvious neurological symptom; 1 as inability to completely extend left anterior claw; 2 as rotation toward the left; 3 as inclining to the left side atwalking; 4 as inability to walk by itself. The higher scale, the more severe the behavioral disturbance was). Brain infarcted area was assayed 1 hour and 23 hours after cerebral ischemia-reperfusion, and was expressed as the percentage of stained area to non-stained area. Neuron apoptosis was detected by the terminal deoxynucleotidyl transferase (TdT)-mediated dUTPfluorescence nick end labeling (TUNEL) method to assess DNA fragments of apoptotic cells 1 hour after cerebral ischemia, and 2 hours, 5 hours and 23 hours after reperfusion. The apoptotic cells were counted under the fluorescence microscope. The percentage of apoptosis cells (%) = (apoptotic neurons ÷ hippocampal neurons) ×100%. The protein expression of caspase-3 was detected by immunohistochemical staining. The positive cells were identified and counted under the light microscope. The percentage of positive cells of caspase-3 (%) = (positive neurons of caspase-3 ÷ hippocampal neurons) ×100%.cells of caspase-3 1 hour after cerebral ischemia, and 2, 5 and 23 hours after reperfusion in each group.scoring: Neurological score of rats in 50 mg/kg, 75 mg/kg Breviscapine groups and MgSO4 group was 1.2±0.4, 0.5±0.4, and 1.3±0.4, respectively,which were all lower than that in IR group [(2.2±0.6)] 23 hours after cerebral ischemia-recirculation (F=6.09, P=0.001). However, it was lower in farcted area in rats of 50 mg/kg and 75 mg/kg Breviscapine groups and MgSO4 group was (0.18±0.03)%, (0.10±0.02)%, and (0.28±0.02)%, respectively, which were all lower than that of IR group [(0.43±0.05) %] 23 hours after cerebral ischemia-reperfusion (F=2.3, P=0.001). It was smaller in hippocampal apoptotic cells: It was (27.2±4.3) %, (20.6±3.6)%, and (35.4±5.5)% in 50 mg/kg and 75 mg/kg Breviscapine groups and MgSO4 group,which were all lower than that of IR group [(60.4±6.2)%] 23 hours after cerebral ischemia-reperfusion (F=6.17, P=0.000 7). It was lower in Brevispositive cells of caspase-3: It was (34.2±5.3)%, (21.6±3.5)%, and (47.4±4.5)% in 50 mg/kgand 75 mg/kg Breviscapine groups and MgSO4 group,which were all lower than that of IR group [(76.3±6.2)%] 23 hours after cerebral ischenia-reperfusion (F=6.88, P=0.000 1). It was lower in Breviscapine group than in MgSO4 group (P ＜ 0.01).CONCLUSION: Breviscapine can protect brain damage induced by cerebral ischemia-reperfusion by markedly decreasing the neurological score,area of cerebral infarction, number of hippocampal apoptotic cells and number of positive cells of caspase-3, and it has better effect than MgSO4.
