缺血再灌注大鼠脑内生长因子的表达及电针任脉的干预作用

目的:观察电针任脉经穴对脑缺血再灌注大鼠脑内碱性成纤维细胞生长因子(bFGF)mRNA、表皮生长因子(EGF)mRNA及神经营养因子(NGF)mRNA表达的影响。方法:采用线栓法建立大鼠大脑中动脉栓塞局灶性脑缺血再灌注模型。72只实验动物随机分为假手术组(Sham)、脑缺血再灌注模型组(MCAO)、模型+电针任脉治疗组(R)。用原位杂交的方法检测脑内侧脑室下区、海马齿状回区bFGFmRNA、EGFmRNA及NGFmRNA的阳性表达。结果:模型组与假手术组相比,3种生长因子的mRNA阳性表达均有不同程度的增加,于造模后7天、14天差异有统计学意义(P<0.05);电针任脉组3种因子的阳性表达均多于模型组,NGFmRNA仅于造模后28天差异有统计学意义(P<0.05),bFGFmRNA及EGFmRNA的阳性表达于造模后14天及28天差异均有统计学意义(P<0.05)。结论:电针任脉经穴能够上调脑缺血再灌注大鼠脑内bFGF、EGF、NGF的表达,这可能是电针任脉促进脑缺血损伤后神经修复的保护机制之一。

应用逆转录病毒pLXSN-EGFP示踪观察电针任脉对MCAO大鼠SVZ区NSC增殖的影响

目的:利用侧脑室注射pLXSN-EGFP在体示踪观察电针任脉对缺血侧SVZ区NSC增殖的情况,探讨电针任脉治疗缺血性脑损伤的作用机制。方法:将病毒上清注入MCAO大鼠侧脑室以标记新增殖细胞,并用Nestin免疫荧光染色法鉴定NSC,并计数不同时相缺血侧SVZ区NSC。结果:脑缺血后不同时相缺血侧SVZ区均有新增殖NSC,且电针任脉组有更明显的增加。结论:缺血可激发SVZ区NSC的增殖,电针任脉可起到明显促进作用,推论这种作用可能是是任脉的妊养和促进脑缺血后神经修复功能的重要机制。

电针任脉经穴对脑缺血再灌注大鼠脑内bFGF蛋白及bFGFm RNA表达的影响

目的:观察电针任脉经穴对脑缺血再灌注大鼠脑内碱性成纤维细胞生长因子(basic fibrob lasticgrowth factor,bFGF)蛋白及mRNA表达的影响。方法:采用线栓法建立大鼠大脑中动脉栓塞局灶性脑缺血再灌注模型。72只实验动物随机分为假手术组(Sham)、脑缺血再灌注模型组(MCAO)、模型+电针任脉治疗组(R)。用免疫组化及原位杂交的方法检测脑内侧脑室下区、海马齿状回区bFGF蛋白及bFGFmRNA的阳性表达。结果:模型组与假手术组相比,bFGF蛋白及bFGFmRNA的阳性细胞数均有不同程度的增加,其中造模后7天、14天差异有统计学意义(P<0.05);电针任脉组的阳性表达多于模型组,但于造模后7天差异无统计学意义(P>0.05),造模后14天及28天差异均有统计学意义(P<0.05)。结论:电针任脉经穴能够上调脑缺血再灌注大鼠脑内bF-GF蛋白及bFGFmRNA表达,这可能是电针任脉促进脑缺血损伤后神经修复的保护机制之一。

电针任脉经穴对MCAO大鼠脑内表皮生长因子表达的影响

目的:观察电针任脉经穴对脑缺血再灌注大鼠脑内表皮生长因子(epidermal growth factor,EGF)蛋白及mRNA表达的影响。方法:采用线栓法建立大鼠大脑中动脉栓塞局灶性脑缺血再灌注模型。72只实验动物随机分为假手术组(Sham)、脑缺血再灌注模型组(MCAO)、模型+电针任脉治疗组(R)。用免疫组化及原位杂交的方法检测脑内侧脑室下区、海马齿状回区EGF蛋白及EGFmRNA的阳性表达。结果:模型组与假手术组相比,EGF蛋白及EGFmRNA的阳性细胞数均有不同程度的增加,其中蛋白表达于再灌注14天恢复至基线水平,而mRNA表达于再灌注28天恢复至基线水平;电针任脉经穴至少能在再灌注7天后上调两区内源性EGF蛋白的表达,而于再灌注14天后上调EGF mRNA的表达,差异有统计学意义(P<0.05)。结论:电针任脉经穴能够上调脑缺血再灌注大鼠脑内EGF蛋白及EGFmRNA表达,这可能是电针任脉促进脑缺血损伤后神经修复的保护机制之一。

Effect of electroacupuncture at conception vessel and governor vessel on correlative signals of stress-injury-repair in cerebral ischemia reperfusion rats

Objective To explore the effect of electroacupuncture at conception vessel and governor vessel on correlative signals of inflammatory response stressinjury-repair in cerebral ischemia reperfusion rats. Methods A total of 216 rats were included in the experiment, of which 192 rats were randomly divided into four groups, including sham operation group（group A）, model group（group B）, governor vessel group（group C）, and conception vessel and governor vessel group（group D）, 48 rats in each group. The rats in each group were further divided into six sub-groups, including 12 h group, 24 h group, 48 h group, 72 h group, 96 h group and 144 h group. MCAO operation course was simulated in group A, and the common carotid artery, internal carotid artery and external carotid artery on the right side were exposed, without insertion of suture or treatment. After completion of modeling in group B, treatment was not conducted. After completion of modeling in group C, ＂Shuǐgōu（水沟 GV 26）＂, ＂Bǎihuì（百会 GV 20）＂ and ＂Dàzhuī（大椎 GV 14）＂ of governor vessel were selected, and electroacupuncture device was connected with ＂GV 14＂ and ＂GV 20＂. After completion of modeling in group D, ＂GV 26＂, ＂GV 20＂ and ＂GV 14＂ of governor vessel, and ＂Qìhǎi（气海 CV 6）＂, ＂Guānyuán（关元 CV 4）＂ and ＂Chéngjiāng（承浆 CV 24）＂ of conception vessel were selected; electroacupuncture device was connected with ＂GV 14＂ and ＂GV 20＂, ＂CV 4＂ and ＂CV 6＂. Neurological severity score was conducted.Enzyme linked immunosorbent assay（ELISA） was applied to determine the concentration of peripheral serum adrenocorticotropic hormone（ACTH） of rats, and immunohistochemical method was adopted to detect the expression levels of tumor necrosis factor α（TNF-α） and transforming growth factor β1（TGF-β1） in brain tissue of rats. The other 24 rats were also randomly divided into four groups mentioned above, 6 rats in each group, to detect cerebral ischemie volume. Results Compared with group B, the ischemia volume in brain tissue at 144 h after cerebral ischemia reperfusion decreased significantly in group C and group D（P〈0.05）, and the ischemia volume in group D was less than that in group C（P〈0.05）. The neurological impairment scores of group C and group D at all time points were lower than those of group B, and the differences were statistically significant（all P〈0.05）. The scores of group D at all time points were lower than those of group C, and the differences since 24 h were statistically significant（all P〈0.05）. Compared with group B, the excessive expression of ACTH was suppressed dynamically and generally in group C and group D（P〈0.05）; the expression level of TNF-α was down-regulated and TGF-β1 was up-regulated as well in group C and group D（P〈0.05）. The effect in group D was superior to that in group C. Conclusion Electroacupuncture at conception vessel and governor vessel can protect brain through the intervention of the expression of correlative signals of inflammatory response stress-injury-repair of cerebral ischemia reperfusion.