经皮穴位电刺激联合全身麻醉或控制性低血压对术后海马神经保护的炎症调节机制

[Inflammatory-modulating mechanism of transcutaneous electrical acupoint stimulation combined with general anesthesia or controlled hypotension on postoperative hippocampal neuroprotection].

作者信息

Liu Zhe, Shen Ying-Jie, Lian Lin-Li, Sha Xiao-Mei, Fang Jian-Qiao

机构信息

Third Clinical Medical College, Zhejiang University of CM, Hangzhou 310053, Zhejiang Province, China.

出版信息

Zhongguo Zhen Jiu. 2013 Feb;33(2):149-55.

PMID:23620945

Abstract

OBJECTIVE

To observe the effects of transcutaneous electrical acupoint stimulation (TEAS) combined with general anesthesia or controlled hypotension on hippocampal neuronal damage and the inflammatory response in peripheral circulation and central nervous system (CNS) after surgery, and to investigate its brain protection mechanism.

METHODS

Eighteen healthy male beagles aged 6 - 8 months were randomly divided into a general anesthesia group (group G), a controlled hypotension group (group C) and a compound anesthesia acupuncture group (group A), 6 cases in each group. Dogs in group G was anesthetized by isoflurane inhalation, and group C was combined with intravenous infusion of sodium nitroprusside based on isoflurane inhalation to induce hypotension, and followed surgery after achieving the target blood pressure, and group A was combined with TEAS at "Quchi" (LI 11), "Hegu" (LI 4) "Zu sanli" (ST 36) and "Sanyinjiao" (SP 6) based on controlled hypotension, and then brain tissue was taken out on the 72 h after mean arterial pressure (MAP) was returned to baseline levels. The concentration of IL-1beta,TNF-alpha in serum at different time points were detected by ELISA. The expression of IL-1beta, TNF-alpha, Bcl-2, Bax and cleaved caspase-3 were measured by immunohistochemistry, and the apoptosis of hippocampus were detected by TUNEL.

RESULTS

(1) At different time points, the concentration of TNFalpha showed the trend of increase first and then decrease, while IL-1beta concentration represented a trend of decrease first and then increase in both group C and group A, but there were no significant differences in cytokine expression between the two groups (all P > 0.05). (2) The ratio of positive cells of IL-1beta, TNF-alpha and caspase-3 in CA1 and CA3 of hippocampus in both group C and A were higher than those in group G (all P < 0.01), and cytokines expression in group A were lower than those in group C (all P < 0.01), and caspase-3 in CA1 in group A was lower than that in group C (P < 0.01). The ratio of Bcl-2/Bax in both group C and A were lower than that in group G (all P < 0.01), and that in group A was higher than that in group C (P < 0.01 in CA1, P < 0.05 in CA3). (3) The apoptosis index (AI) of hippocampal neurons in both group C and A was significantly higher than that in group G (P < 0.01), while AI in CA1 in group A was lower than that in group C (P < 0.01).

CONCLUSION

The TEAS can regulate the expression of inflammatory factor in hippocampus in animals undergoing general anesthesia or con trolled hypotension surgery, further improving Bcl-2/Bax ratio, inhibiting the expression of caspase-3 and reducing neuron apoptosis in hippocampus so as to play a neuroprotection.

摘要

目的

观察经皮穴位电刺激（TEAS）联合全身麻醉或控制性低血压对术后海马神经元损伤及外周循环和中枢神经系统（CNS）炎症反应的影响，并探讨其脑保护机制。

方法

将18只6 - 8月龄健康雄性比格犬随机分为全身麻醉组（G组）、控制性低血压组（C组）和复合麻醉针刺组（A组），每组6只。G组犬采用异氟烷吸入麻醉，C组在异氟烷吸入基础上联合静脉输注硝普钠诱导低血压，达到目标血压后进行手术，A组在控制性低血压基础上联合在“曲池”（LI 11）、“合谷”（LI 4）、“足三里”（ST 36）和“三阴交”（SP 6）进行TEAS，然后在平均动脉压（MAP）恢复至基线水平后72小时取出脑组织。采用酶联免疫吸附测定（ELISA）法检测不同时间点血清中白细胞介素-1β（IL-1β）、肿瘤坏死因子-α（TNF-α）的浓度。采用免疫组织化学法检测IL-1β、TNF-α、Bcl-2、Bax和裂解型半胱天冬酶-3的表达，采用末端脱氧核苷酸转移酶介导的缺口末端标记法（TUNEL）检测海马凋亡情况。

结果

（1）在不同时间点，C组和A组TNF-α浓度均呈先升高后降低趋势，IL-1β浓度呈先降低后升高趋势，但两组细胞因子表达差异均无统计学意义（均P > 0.05）。（2）C组和A组海马CA1区和CA3区IL-1β、TNF-α和半胱天冬酶-3阳性细胞比例均高于G组（均P < 0.01），A组细胞因子表达低于C组（均P < 0.01），A组CA1区半胱天冬酶-3低于C组（P < 0.01）。C组和A组Bcl-2/Bax均低于G组（均P < 0.01），A组高于C组（CA1区P < 0.01，CA3区P < 0.05）。（3）C组和A组海马神经元凋亡指数（AI）均显著高于G组（P < 0.01），而A组CA1区AI低于C组（P < 0.01）。