参附注射液对脑再灌注损伤中细胞凋亡的影响

Effect of Shenfu injection on brain apoptosis and expression of HO-1 during focal cerebral ischemia reperfusion injury in rats

目的探讨参附注射液（Shenfuinjection，SF）对大鼠局灶性脑缺血-再灌注损伤（cerebralischemiareperfusioninjury，CIRI）中脑神经元细胞凋亡及血红素氧合酶-1（hemeoxygenase-1，HO-1）表达的影响，并探讨其可能的机制。方法SD雄性大鼠42只，随机（随机数字法）分为假手术组（Sham组）、缺血-再灌注组（IR组）、参附注射液组（SF组），制造大脑中动脉阻塞模型，缺血2h后再灌注3h、6h。SF组于再灌注时腹腔注射参附注射液10ml／kg。光镜和电镜下观察脑组织神经细胞形态学变化，用Annexin-V—PI双染法通过流式细胞仪检测脑织凋亡细胞并计算凋亡率，应用RT-PCR技术检测HO-1。结果IR组与Sham组相比，3、6h时点脑组织凋亡率明显增加（P〈0．01）。SF组与IR组相比，SF组3、6h时点脑织凋亡率明显减少，差异均具有统计学意义（P〈0．01），3、6h时点脑组织HO-1的表达显著增加（P〈0．01），脑组织结构损害明显减轻。结论早期应用参附注射液可以诱导HO-1大量表达从而抑制细胞凋亡的发生，进而减轻脑组织损伤。

Objective To investigate the effects of Shenfu injection （SF, a Chinese herbal medicine preparation made of Codonopsis pilosula and Aconitum carmichaeli） on the cell apoptosis of focal cerebral ischemic-reperfusion injured rats and the expression of heine oxygenase-1 （ HO-1 ）. Methods Forty-two male Sprague-Dawley rats used for producing unilateral brain ischemia reperfusion model were randomly divided into three groups： sham operation group （Sham group） , ischemia reperfusion group （IR group ） , and SF Injection group （ SF group）. The model of focal cerebral ischemia-reperfnsion injury was induced by transient occlusion of middle cerebral artery （ischemia for 2 h, and reperfusion for 3, 6 h respectively）. In SF group, SF （ 10 mg/kg） was intraperitoneally injected during reperfusion. Cell apoptosis rate in brain tissue was detected by the technique of Annexin-V-PI double staining and was counted in flow cytometer. Expression of HO-1 in brain was measured by RT-PCR, while the pathological and ultra structure changes of cerebral tissue were also observed. Results Cell apoptosis rate of brain tissue were significantly higher in IR group than that in Sham group （ P 〈 0. O1 ）, while SF group had less significant changes in cell apoptosis rate, HO-1 level of brain tissue than IR group （P 〈0. 01 ）. The ultra structure change of brain tissue was less in SF group than that in IR group. Conclusions During early stage of brain IR injury, SF inhibits cellular apoptosis and in turn protects the brain from injury which is attributed to the increase in HO-1 expression induced by SF.