Electro-acupuncture decreases hydrogen sulfide concentration in brain cortex of baby rats with hypoxic-ischemic brain damage

目的 建立缺氧缺血性脑损伤(HIBD)大鼠模型,研究电针干预后大鼠大脑皮层硫化氢(H2S)含量及相关酶的表达变化,探讨电针治疗神经系统疾病的气体生物学机制.方法 SD大鼠32只,日龄7 d,随机分为4组,假手术(sham)对照组,sham+电针(EA)组,HIBD对照组和HIBD+EA组,每组8只.Sham+EA组、HIBD+EA组大鼠于造模次日开始予以电针刺激百会、大椎穴,30min/d,14 d为1疗程.对照组仅固定四肢,不予针刺.疗程结束次日将大鼠处死、取材.应用敏感硫电极法测定大脑皮层H2S含量;应用免疫组织化学法定位和半定量检测胱硫醚-β-合成酶(cystathionine-β-synthase,CBS)表达;应用蛋白质免疫印迹技术定量分析大脑皮层组织CBS表达.结果 HIBD对照组幼鼠脑皮层H2S含量为(26.83±4.31)nmol/mg蛋白,较sham对照组[(22.78±1.54)nmol/mg蛋白]明显升高,差异有统计学意义(P＜0.01);HIBD+EA组幼鼠脑皮层H2S含量为(18.08±2.71)amol/mg蛋白,sham+EA组为(18.91±2.78)amol/mg蛋白,较相应对照组明显降低,差异有统计学意义(P＜0.01);EA组大鼠脑皮层CBS表达较相应对照组明显降低,差异有统计学意义(P＜0.05).结论 电针可下调大脑皮层H2S/CBS体系,这可能是电针发挥对脑损伤修复作用的机制之一.Objective To establish hypoxic-ischemic brain damaged (HIBD) rat model,investigate whether H2S and cystathionine-β-synthase (CBS), the key enzyme for its generation, may be a mediator of electro-acupuncture(EA) stimulation treatment for HIBD. Methods Thirty-two healthy Sprague-Dawley neonatal rats were divided into four groups randomly: sham control group ( n = 8 ); sham + EA group ( n =8); HIBD control group ( n = 8); and HIBD + EA group ( n = 8 ). HIBD rat models were established on their 7-day-old. From the next day ,rats of sham + EA group and HIBD + EA group were electric stimulated 30 min daily for 14 d,BAIHUI and DAZHUI as the acupoints. Control ones were just fixed at the same time,without acupuncture. The rats were sacrificed on the 22 nd day, one day after the treatment course. Cortical H2S concentration was measured by sensitive sulphur electrode assay. The CBS protein expression was measured by western blot analysis and immunohistochemistry for localization. Results The concentration of cortical H2S in HIBD control group was (26. 83 ± 4. 31 ) nmol/mg protein, which was significantly higher than that of sham control group[(22. 78 ± 1.54) nmol/mg protein]( P ＜ 0. 01 ). The H2 S levels in HIBD + EA group and sham + EA group were ( 18.08 ± 2.71 ) nmol/mg protein and ( 18.91 ± 2. 78 ) nmol/mg protein, respectively. Compared with corresponding control group, they were much lower( P ＜ 0. 01 ). The expression of CBS protein in rats with EA stimulation decreased in cortex compared to corresponding control group( P ＜0. 05 ).Conclusion EA can down-resulate H2S/CBS pathway. This may be one of the mechanisms of how EA contributes to the recovery of brain damage.北京市科委国际合作项目; 国家科委国际合作项目中国科技核心期刊(ISTIC)02142-1451