电针对大脑中动脉梗塞大鼠EphrinB2/EphB2信号通路上突触重塑相关因子的影响

目的:观察电针对大脑中动脉梗塞模型(MCAO)大鼠促红细胞生成素产生肝细胞配体-B2(erythropoietin-producing hepatocyte receptor interacting protein-B2,Ephrin B2)/受体-B2(erythropoietin-producing hepatocyte receptor-B2,Eph B2)信号通路上突触重塑相关因子表达的影响,以期揭示针刺治疗脑缺血的可能机制。方法:120只雄性SD大鼠随机分为假手术组、模型组、穴位组、非穴位组(n=30);各组再分为术后3d,14d及21d三个亚组(n=10)。除假手术组外,其余各组大鼠用Zea-longa线栓法制备MCAO模型;穴位组和非穴组电针治疗,其余两组只绑不针。对大鼠行神经功能评分,采用免疫组化、免疫荧光和蛋白质印迹法行相应指标检测。结果:模型组术后3d梗死灶周Ephrin B2/Eph B2表达明显减少,但随时间呈递增趋势,21d恢复到假手术水平。穴位组较模型组和非穴组增加更明显,术后21d表达高于假手术组(P0.05)。Ras相关的C3肉毒毒素底物1(ras-related C3 botulinum toxin substrate-1,Rac-1)和微管相关蛋白-2(microtubule-associated protein-2,MAP-2)的表达随时间逐渐递增,且均与神经功能改善相吻合;其中穴位组增加最明显,术后21d超假手术组水平(P0.05)。结论:Ephrin B2/Eph B2信号通路参与了MCAO大鼠突触重塑,电针肝俞、肾俞穴可能通过激活Ephrin B2/Eph B2信号通路,促进MCAO大鼠神经功能的康复。     

艾灸治疗胃黏膜损伤研究述评

艾灸在治疗胃黏膜损伤性疾病临床及实验研究中取得了一定的进展,同时也暴露出了研究中的不足之处:1治疗机制研究尚不完善。艾灸胃相关穴位可促进胃黏膜保护因子及抗凋亡蛋白的含量增加,抑制损伤因子及凋亡蛋白的产生,从而防治胃黏膜损伤。体液、内分泌、免疫等也是艾灸发挥作用的途径之一,其如何共同协作形成网络系统,发挥巨大疗效机制尚不完善,有待深度观察与研究。2关于艾灸对胃黏膜的保护作用机制尚在细胞因子、蛋白层面,鲜有关于基因改变方面的研究,今后可在此基础上继续发展,为治疗胃黏膜损伤性疾病提供新思路与新途径。3在灸量方面,虽有不少学者对其有所研究,提出影响艾灸量效关系因素,却无治疗有效病症的最佳灸量标准。今后临床中可进行关于不同人群、何种灸量可达到某种治疗效果方面的研究,从而总结出治疗不同病因造成胃黏膜损伤的最佳灸疗方案。4灸法方面,虽然方法众多,与其他疗法比较疗效确切,对各种灸法之间的比较很少,不同灸法之间疗效特异性不显著,不利于各种灸法之间灸效的判断及临床应用的选择,因而今后在研究中可针对这几个方面进行探索。     

Effect of electroacupuncture on expressions of VEGF and CD31 in MCAO model rats

Objective

To investigate the effect of electroacupuncture (EA) at Ganshu (BL 18) and Shenshu (BL 23) on vascular endothelial growth factor (VEGF) and platelet endothelial cell adhesion molecule-1 (PECAM-1)/CD31 around the cerebral infarction focus in middle cerebral artery occlusion (MCAO) rats and the possible mechanism, thus to provide a new strategy for the treatment of cerebral ischemic stroke by acupuncture.

Methods

A total of 180 healthy male Sprague-Dawley (SD) rats were randomly divided into a sham operation group, a model group, an acupoint group and a non-acupoint group, 45 rats in each group. MCAO model was established using the modified line-embolus method in all rats except for those in the sham operation group; rats in the acupoint group were treated with EA at Ganshu (BL 18) and Shenshu (BL 23); rats in the non-acupoint group were treated with EA at the control points; rats in other 2 groups were only subjected to bundling without treatment. Ten rats in each group were randomly selected on the 3rd day, the 14th day and the 21st day after acupuncture stimulation to test the neurological function impairment. The expression levels of CD31 and VEGF were also detected.

Results

Compared with the model group and non-acupoint group, the neurological function score of the acupoint group was decreased at each time point, and the differences were statistically significant (P<0.05, P<0.01). The expressions of VEGF and CD31 in each group were the lowest on the 3rd day, reached the peak on the 14th day and still remained at high level on the 21st day. And the differences among groups were statistically significant both on the 14th day and the 21st day (P<0.05, P<0.01). Compared with the model group and the non-acupoint group, the expressions of VEGF and CD31 in the acupoint group were increased, and the differences were statistically significant (all P<0.05).

Conclusion

EA at Ganshu (BL 18) and Shenshu (BL 23) can significantly improve the neurological function score of MCAO model rats, and shows protective effect on cerebral ischemia. The protective mechanism may be related to the up-regulation of CD31 and VEGF expression around the cerebral infarction focus in the MCAO model rats and induction of angiogenesis.

     

Effect of electroacupuncture at Ganshu (BL 18) and Shenshu (BL 23) on the expression of EphB2 protein in cortex around cerebral infracted area of rat

Objective

To observe the effect of electroacupuncture (EA) on the expression of erythropoie-tin-producing hepatocyte receptor B2 (EphB2) in the cortex around the infracted area of middle cerebral artery occlusion (MCAO) rats at different timing, and to reveal the possible mechanism of acupuncture in the treatment of cerebral ischemia.

Methods

A total of 180 male Sprague-Dawley (SD) rats were randomly divided into a sham operation group, a model group, an acupoint group and a non-acupoint group, with 45 rats in each group. Rats in each group were further divided into three subgroups: postoperative 3 d, postoperative 14 d and postoperative 21 d groups, with 15 rats in each subgroup. The MCAO model was made by the modified occlusion method. The neurological function score, 2,3,5-triphenyl tetrazolium chloride (TTC) staining, immunohistochemistry assay, immunofluorescence double labeling method and Western blot were used to detect the corresponding indicators.

Results

The neurological impairment of rats was most obvious at postoperative 3 d, and then gradually improved with time, which was more significant in the acupoint group (P<0.05). The change of infarcted volume was consistent with the neurological function impairment. The number of EphB2 positive cells (EphB2⁺) around the infarcted area was decreased significantly at postoperative 3 d, and then gradually improved with time, which returned to the same level as that in the sham operation group at postoperative 21 d. The increase was most significant in the acupoint group (P<0.05), and the positive cell number was higher than that in the sham operation group (P<0.01). Western blot and immunohistochemistry results were basically consistent. Immunofluorescence displayed that EphB2⁺ and postsynaptic density-95 positive (PSD-95+) were co-expressed, after the MCAO operation, in the cortical neuron around the infracted area, and the number of co-expressing cells was increased gradually with time, which was most significant in the acupoint group (P<0.05).

Conclusion

Electroacupuncture at Ganshu (BL 18) and Shenshu (BL 23) can significantly improve the neurological function and cerebral infarcted volume ratio of MCAO rats, which may be related to the activation of EphB2 expression in cortex around the infracted area and the promotion of synaptic remodeling.

     

魏玉霞应用大柴胡汤异病同治经验探析

大柴胡汤是《伤寒杂病论》中表里双解代表方剂之一，在临床上应用比较广泛，涉及领域众多，少阳不和兼有阳明里实是其病机关键所在。魏玉霞主任医师从事临床诊疗多年，经验丰富，擅长运用大柴胡汤加减治疗便秘、郁证等以肝胆郁滞、热结胃肠为基本病机的多种疾病，临床疗效颇佳。