电针对高脂血症合并脑缺血损伤大鼠海马区钙结合蛋白D28K蛋白表达的影响

目的:观察针刺对高脂血症合并脑缺血损伤大鼠大脑海马区细胞凋亡及钙结合蛋白(Calbindin)D28 K(CB)表达的影响。方法:SD雄性大鼠,随机分为正常对照组、模型组、治疗Ⅰ组和治疗Ⅱ组,每组10只。采用经典高脂配方饲料喂养并结合FeCl3化学刺激诱导血栓性闭塞大脑中动脉,造成大鼠高脂血症合并脑缺血模型。其中治疗Ⅰ组先电针"三阴交""丰隆",每日1次,治疗10 d后再造脑缺血模型;治疗Ⅱ组在造模成功后再开始治疗。两治疗组造模完成后均电针"三阴交""丰隆",手针"百会""水沟",每日1次,治疗7 d。应用免疫组化方法观察大鼠海马区CB蛋白表达的变化,原位末端标记染色法观察海马区神经细胞的凋亡情况。结果:高脂血症大鼠局灶性脑缺血后细胞凋亡率较正常组明显增高,CB阳性细胞较正常组明显减少,阳性强度也减弱(P0.05);针刺后各治疗组CB阳性细胞的表达明显增加,其中治疗Ⅰ组较治疗Ⅱ组增加显著(P0.05),同时各治疗组神经细胞凋亡亦明显减少,以治疗Ⅰ组较为明显(P0.05)。结论:针刺可减少高脂血症大鼠海马区神经细胞的凋亡,增加CB的表达,减轻脑缺血损伤程度。

Effect of Electroacupuncture on the Expression of Hippocampal Calbindin-D 28 K in Hyperlipemia Rats with Concurrent Cerebral Ischemia

Objective To observe the effect of electroacupuncture(EA)on Calbindin D 28 K(CB)expression and apoptosis in hippocampus in hyperlipemia rats with concomitant cerebral ischemia(Cl).Methods Forty male SD rats were randomized into control,hyperlipemia+Cl(model),acupuncture Ⅰ,and acupuncture Ⅱ groups,with 10 cases in each.Hyperlipemia model was established by feeding the animals with high fat forage for 6 weeks and Cl model established by occlusion of the unilateral middle cerebral artery.For rats of acupuncture Ⅰ group,EA(1-3 mA,15 Hz)was applied to bilateral "Sanyinjiao"(SP 6)and "Fenglong"(ST 40)for 20 min every time,once daily for 10 days before Cl.For rats of both acupuncture Ⅰ and Ⅱ groups,after Cl,EA was applied to SP 6,ST 40,"Baihui"(GV 20)and "Shuigou"(GV 26)were punctured with filiform needles and stimulated by twirled the needle with hand continuously for 1 min,once daily for 7 days.The apoptotic cells of hippocampal CA 3 area were displayed by TUNEL(terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling)method.CB antibody staining with immunohistochemistry was used to show CB-positive cells which were observed by optic microscope.Results In comparison with control group,the percentage of apoptotic cells of the hippocampal CA 3 area in model group increased considerably(P＜0.01),while the number of CB immunoreaction(IR)-positive cells in the same area decreased significantly(P＜0.05).Compared with model group,the percentages of apoptotic cells of CA 3 area in acupuncture Ⅰ and acupuncture Ⅱ groups lowered remarkably(P＜0.01),while the numbers of CB IR-positive cells in the later two groups increased significantly(P＜0.01).The therapeutic effect of acupuncture Ⅰ group was apparently superior to that of acupuncture Ⅱ group in lowering the percentage of apoptotic cells and upregulating CB IR-positive cell number(P＜0.05).Conclusion Acupuncture can effectively lessen neuronal apoptosis of the hippocampus CA 3 area which may be closely related to its effect in upregulating the expression of CB in hyperlipemia+Cl rats.Moreover,the cerebral ischemia injury degree may be lightened by acupuncture-lowering hyperlipemia first.