健脑安对血管性痴呆大鼠海马区IL-1β诱导的c-fos蛋白表达的影响

目的 :观察健脑安 (CGE)对脑缺血 再灌注大鼠高表达的IL-1β ,c-fos蛋白的抑制作用。 方法 :采用左侧大脑中动脉插入丝线结扎方法制造大鼠血管性痴呆模型 ,分成健脑安大 (19.34× 10³ g·L^-1生药 )、中 (9.67× 10 ³ g·L^-1生药 )、小 (4.83× 103 g·L^-1生药 )剂量组、IL-1ra对照组、假手术组和模型组。中药用 0 .5 %羧甲基纤维素 (CMC)配成水溶液 ,每 10 0g大鼠灌胃给中药溶液 0 .7mL。IL 1ra对照组大鼠在缺血前 30min和缺血后 10min左侧脑室内分别注射rhIL-1ra-10 μg,假手术组注射同等剂量生理盐水 10 μL。假手术组、模型组分别于造模 7d前开始给予中药灌胃 ,分别按照缺血再灌后不同时间点 (0 .5 ,1,2 ,3,4 ,6 ,8,12 ,24 ,4 8,72 ,96 ,144 ,168h)取材。应用免疫组化方法标记实验各组大鼠应用健脑安和IL-1ra后脑组织中海马区IL-1β蛋白和c-fos阳性神经元的数量 ,进行组间均数比较。 结果 :与模型组相比 ,健脑安 3个剂量组在不同再灌时间 (2 ,3,4 ,12 ,72h)均具有显著性的抑制IL-1β ,c-fos蛋白表达的作用 ,但小、中剂量组的抑制水平不及IL-1ra(P <0.05和P <0.01)。健脑安抑制海马IL-1β蛋白表达 ,2h起效 ,大剂量 (531± 151.1)和中剂量 (589.3± 78.6 )都强于模型组 (687.6±116.7)(P<0.01和0.05),直到72h。大剂量组从缺血-再灌注4h后对c-fos 蛋白表达的抑制作用（81.3±16.1)与IL-lra对照组（67.2±25.7）无显著性差异。结论：具有益气补肾、活血化痰作用的健脑安可以抑制血管性痴呆大鼠脑内IL-1β-fos蛋白的表达亢进，但这一作用起效相对IL-lra慢了大约1h，且与剂量有关.

The influence of CGE on expression of IL-1β and c-fos protein in the hippocampus region in rats following cerebral ischemia-reperfusion

OBJECTIVE: To observe inhibiting effect of CGE (compound ginseng extract) on increased expression of IL-1beta and c-fos protein following cerebral ischemia-reperfusion. METHOD: The vascular dementia model was made by middle cerebral artery occlusion for 2 hours. Expression of IL-1beta and c-fos were determined by immunohistochemistry in the hippocampus regions in brain tissue at the 0.5 h-7 d after reperfusion. CGE was diluted by CMC and poured into the stomach by 0.7 mL x (100 g)(-1) with a high dosage (19.34 x 10(3) g x L(-1) row herbs), a middle dosage (9.67 x 10(3) g x L(-1)), a low dosage (4.83 x 10(3) g x L(-1)). There were an IL-1ra (rhIL-1ra 20 microg injected into the left cerebral ventricle), a sham operation (NaCl 20 microL injected into the left cerebral ventricle) and a model as control. RESULT: Compared with control group, three dose groups (low, middle and high) in CGE showed significant inhibiting effects on the expression of c-fos protein at 2, 3, 4, 12 hours and 3 day following cerebral ischaemic-reperfusion. The level of the inhibiting effects in small and middle groups were lower at all time points than that in IL-1ra group (P < 0.05 to P < 0.01). CGE inhibited the expression of hippocampus IL-1beta protein, taking effect from the 2 h after reperfusion. Both HD group (531 +/- 151.1) and MD group (589.3 +/- 78.6) showed more obvious effect which lasted until the 72 h compared with the model group (687.6 +/- 116.7) (P < 0.01 and 0.05). Large dose group (81.3 +/- 16.1) showed the same level of the inhibiting effect on expression of c-fos protein as IL-1ra group (67.2 +/- 25.7) from 4 hour on following cerebral ischaemic reperfusion (P > 0.05). CONCLUSION: CGE with function of Yiqi Bushen, Huoxue Huatan has effect of inhibiting up-regulated expression of IL-1beta and c-fos protein following cerebral ischemia-reperfusion. However, this effect of CGE starts relatively later than that of IL-1ra. The effect of CGE is associated with its dosage, i.e. a larger dosage has a better effect on expression of c-fos protein in post-stroke dementia.