针刺调控外周血Th1/Th2细胞平衡改善血管性痴呆大鼠脑内神经炎性反应

Acupuncture for improving neuroinflammation in vascular dementia rats by regulating Th1/Th2 cells balance in peripheral blood

目的:观察针刺对血管性痴呆(VD)大鼠外周血辅助型T(Th)1/Th2细胞平衡、炎性反应及脑内神经炎性反应的影响,探讨针刺改善VD认知功能的机制。方法:将60只SPF级Wistar大鼠随机分为正常组(12只)、假手术组(12只)和手术组(36只)。手术组采用双侧颈总动脉结扎术制备VD大鼠模型,再将造模成功的大鼠随机分为模型组和针刺组,每组12只。针刺组予“三焦针法”进行针刺干预,穴取“膻中”“中脘”“气海”“血海”“足三里”,每穴行针30 s,不留针,每日1次,共15 d,第8天休息1次。采用Morris水迷宫进行行为学检测;流式细胞术检测外周血Th1/Th2细胞比值;Luminex液相芯片检测血清及海马白介素(IL)-4、IL-10、干扰素-γ(IFN-γ)、肿瘤坏死因子-α(TNF-α)水平。结果:正常组与假手术组各项指标比较差异无统计学意义(P>0.05)。与假手术组比较,模型组大鼠隐蔽平台实验及反向平台实验逃避潜伏期延长(P<0.01);空间探索实验原平台象限停留时间缩短、穿越原平台次数减少(P<0.01,P<0.05);外周血Th1细胞比例升高、Th2细胞比例下降、Th1/Th2细胞比值增大(P<0.01);血清及海马TNF-α及IFN-γ水平升高、IL-4及IL-10水平降低(P<0.05,P<0.01)。与模型组比较,针刺组大鼠隐蔽平台实验及反向平台实验逃避潜伏期缩短(P<0.01);空间探索实验原平台象限停留时间延长(P<0.05);外周血Th1细胞比例下降、Th2细胞比例升高、Th1/Th2细胞比值减小(P<0.05);血清及海马IFN-γ及TNF-α水平降低、IL-4及IL-10水平升高(P<0.05,P<0.01)。结论:针刺能改善VD大鼠认知功能障碍,降低脑内神经炎性反应水平,其机制可能与调控外周血Th1/Th2细胞平衡、减轻外周炎性反应有关。

Objective To observe the effect of acupuncture on the balance of T helper(Th)1/Th2 cells in peripheral blood,inflammatory reaction and intracerebral neuroinflammation in vascular dementia(VD)rats,and to explore the mechanism of acupuncture for improving cognitive function in VD.Methods A total of 60 SPF Wistar rats were randomized into a normal group(n=12),a sham operation group(n=12)and an operation group(n=36).Bilateral common carotid artery occlusion was adopted to establish the VD model in rats of the operation group.The rats of successful modeling were randomized into a model group and an acupuncture group,12 rats in each one.In the acupuncture group,Sanjiao acupuncture was applied at"Danzhong"(CV 17),"Zhongwan"(CV 12),"Qihai"(CV 6),"Xuehai"(SP 10)and"Zusanli"(ST 36),the needles were manipulated for 30 s at each acupoint,without retaining.The intervention was given once a day for 15 days,and there was 1-day rest on day 8.Morris water maze test was adopted to observe the ethology,flow cytometry was used to detect the ratio of Th1/Th2 in peripheral blood,and Luminex liquid chip technology was used to detect the levels of interleukin(IL)-4,IL-10,interferon-γ(IFN-γ)and tumor necrosis factor-α(TNF-α)in serum and hippocampus.Results There were no significant differences in various indexes between the normal group and the sham operation group(P>0.05).Compared with the sham operation group,in the model group,the escape latency of hidden platform test and reversal platform test was prolonged(P<0.01),the residence time of the original platform quadrant was shortened and the number of crossing the original platform was reduced in probe test(P<0.01,P<0.05),the proportion of Th1 cells was increased,the proportion of Th2 cells was decreased and the ratio of Th1/Th2 cells was increased in peripheral blood(P<0.01),the levels of TNF-αand IFN-γwere increased,the levels of IL-4 and IL-10 were decreased in serum and hippocampus(P<0.05,P<0.01).Compared with the model group,in the acupuncture group,the escape latency of hidden platform test and reversal platform test was shortened(P<0.01),the residence time of the original platform quadrant of the probe test was prolonged(P<0.05),the proportion of Th1 cells was decreased,the proportion of Th2 cells was increased and the ratio of Th1/Th2 cells was decreased in peripheral blood(P<0.05),the levels of TNF-αand IFN-γwere decreased,the levels of IL-4 and IL-10 were increased in serum and hippocampus(P<0.05,P<0.01).Conclusion Acupuncture can improve the cognitive dysfunction and reduce the intracerebral neuroinflammation in VD rats,its mechanism may relate to the regulation of Th1/Th2 cells balance and reduce the inflammatory reaction in peripheral blood.