Wang Zhuo-Yu, Ma Jia-Jia, Guan Han-Yu, Tian Yao, Ren Xiu-Jun, Ma Hui-Fang
School of Acu-moxibustion and Tuina, Beijing University of Chinese Medicine, Beijing 100029, China.
Zhen Ci Yan Jiu. 2017 Apr 25;42(2):119-24.
To observe the effect of electroacupuncture (EA) stimulation of "Fenglong" (ST 40), "Sanyinjiao" (SP 6) plus manual acupuncture (MA) stimulation of "Shuigou" (GV 26) and "Baihui" (GV 20) on Caspase-3 protein expression in the cerebral cortex of rats with hyperlipemia and cerebral ischemia(HL-CI),so as to reveal its mechanisms underlying improvement of HL-CI.
Forty-five rats were randomly divided into normal control,sham operation,model,EA group I(EA+MA was given for 14 days, i.e., 7 days before CI, and 7 days more after HL-CI)and EA group Ⅱ (EA+MA was given for only 7 days after HL-CI),with 9 rats being in each group. The HL-CI model was established by feeding the animals with high fat forage for 6 weeks and then making an occlusion of the unilateral middle cerebral artery by regional application of quantitative paper adsorbing 50% FeCl solution (10 μL). Rats of the sham operation group were treated with the same procedures only without application of FeCl solution. For rats of the EA group I,EA (1-3 mA, 2 Hz/100 Hz) was applied to bilateral acupoints SP 6 and ST 40 (for 20 min),and MA stimulation applied to GV 26 and GV 20. EA was conducted once daily for 7 days after 6 weeks' high fat fo-rage feeding, and EA+MA intervention was conducted once daily for 7 days after CI modeling. For rats in the EA group Ⅱ, EA+MA was applied to the same 4 acupoints once a day for 7 days only after CI modeling. The neurological impairment was assessed by Zea Longa's scoring. The blood sample was taken from the abdominal aorta for measuring the contents of serum cholesterol (CHO),triglyceride (TG) and high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C). Pathological changes of the cerebral cortex were observed after H.E. staining, and the expression of cerebro-cortex Caspase-3 was analyzed by immunohistochemistry.
Following modeling,the neurological score,CHO, TG and LDL-C contents, and the number of Caspase-3 positive cells as well as Caspase-3 immunoactivity level were significantly increased in the model group(<0.05), while serum HDL-C level was obviously decreased(<0.05). After the treatment,the increased neurological score, CHO, TG and LDL-C contents, and the number of Caspase-3 positive cells and Caspase-3 immunoactivity level were considerably decreased in the EA group I and Ⅱ(<0.05)while the decreased HDL-C level was notably increased relevant to the model group(<0.05). The effects of the EA group I were notably superior to those of EA group Ⅱ in down-regulating the neurological score,CHO, TG and LDL-C levels and the expression of Caspase-3 protein(<0.05). No significant differences were found between the normal control and sham operation groups in the neurological scores 20 min and 7 days after modeling and Caspase-3 expression levels (>0.05). H.E. staining showed a reduction of the apoptotic cells and inflammatory cells in both EA group I and Ⅱ.
Both EA and EA+MA interventions can improve neurological function in HL-CI rats,which may be related to their effects in adjusting the levels of serum lipids and down-regulating the expression of cell apoptosis-related Caspase-3 protein in the ischemic cortex. Moreover, the cerebral ischemia injury may be lightened by EA-lowering hyperlipemia first.
观察电针(EA)刺激“丰隆”(ST 40)、“三阴交”(SP 6)加手针(MA)刺激“水沟”(GV 26)和“百会”(GV 20)对高脂血症合并脑缺血(HL-CI)大鼠大脑皮质Caspase-3蛋白表达的影响,以揭示其改善HL-CI的作用机制。
45只大鼠随机分为正常对照组、假手术组、模型组、电针Ⅰ组(造模前7天及造模后高脂血症合并脑缺血7天给予EA+MA,共14天)和电针Ⅱ组(高脂血症合并脑缺血后仅给予7天EA+MA),每组9只。采用高脂饲料喂养动物6周,然后用定量滤纸蘸取50%FeCl3溶液(10 μL)局部应用于大鼠单侧大脑中动脉造成闭塞,建立HL-CI模型。假手术组大鼠除不应用FeCl3溶液外,其余操作同模型组。电针Ⅰ组大鼠,双侧穴位SP 6和ST 40给予电针(1~3 mA,2 Hz/100 Hz,20 min),GV 26和GV 20给予手针刺激。高脂饲料喂养6周后电针治疗7天,每天1次;造模后给予电针+手针干预7天,每天1次。电针Ⅱ组大鼠仅在造模后对相同4个穴位给予电针+手针干预,每天1次,共7天。采用Zea Longa评分法评估神经功能缺损。经腹主动脉取血,检测血清胆固醇(CHO)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)和低密度脂蛋白胆固醇(LDL-C)含量。HE染色后观察大脑皮质病理变化,免疫组织化学法分析大脑皮质Caspase-3的表达。
造模后,模型组神经功能评分、CHO、TG和LDL-C含量、Caspase-3阳性细胞数及Caspase-3免疫活性水平均显著升高(P<0.05),血清HDL-C水平明显降低(P<0.05)。治疗后,电针Ⅰ组和Ⅱ组神经功能评分、CHO、TG和LDL-C含量、Caspase-3阳性细胞数及Caspase-3免疫活性水平均显著降低(P<0.05),血清HDL-C水平较模型组显著升高(P<0.05)。电针Ⅰ组在下调神经功能评分、CHO、TG和LDL-C水平及Caspase-3蛋白表达方面的效果显著优于电针Ⅱ组(P<0.05)。造模后20 min及7天,正常对照组和假手术组神经功能评分及Caspase-3表达水平差异无统计学意义(P>0.05)。HE染色显示电针Ⅰ组和Ⅱ组凋亡细胞和炎症细胞均减少。
电针及电针+手针干预均可改善HL-CI大鼠神经功能,可能与其调节血脂水平及下调缺血皮质细胞凋亡相关Caspase-3蛋白表达有关。此外,电针先降低高脂血症可能减轻脑缺血损伤。