电针刺对创伤大鼠脾T细胞白细胞介素-2、γ干扰素及细胞外信号调节激酶通路的影响  

Effects of electroacupuncture on interleukin -2 , interferon -gamma and extracellular-regulated protein kinase signaling pathways in the splenic T cells of traumatized rats

作　　者：王坤[1] 张健[1] 徐杨[1] 王玉[1] 王国年[1] 

机构地区：[1]哈尔滨医科大学附属第三医院麻醉科,150081

出　　处：《国际麻醉学与复苏杂志》2012年第2期76-79,125,共5页International Journal of Anesthesiology and Resuscitation

摘　　要：目的观察电针刺（electmacupuncturelEA）对手术创伤大鼠脾脏T细胞白细胞介素-2（interleukin-2，IL-2）、γ干扰素（Interferon-gamma，IFN-γ）和细胞外信号调节激酶1／2（extracellular—regulated protein kinasel／2，ERK1／2）信号通路的影响，探讨其免疫调节的作用机制。方法32只Sprague-Dawley（SD）大鼠用随机数字表法随机分为两组（每组8只）：对照组（Control组）、单纯EA组（EA组）、创伤组Trauma组）和创伤后EA组（T＋EA组）。大鼠麻醉后，沿背部中线、腹部中线行6cm和5em的纵向切口，暴露腹部脏器3min，制作免疫抑制模型。选双侧“足三里”和“阑尾”穴行EA。于术后3d无菌取脾，尼龙毛柱法分离脾T淋巴细胞用酶联免疫吸附试验（ELISA）和逆转录聚合酶链反应（reverse transcription-polymerase chain reaction，RT-PCR）法分别检测脾脏T淋巴细胞IL-2、IFN-γ蛋白和mRNA表达；应用Western-blot检测ERKl／2表达；核转录因子_I（B（nuclearfactor．KB，NF-KB）和活化蛋白-1（activatorprotein-1，AP-1）活性测定采用Trans-AMELISA-basedkits分析法。结果Trauma组脾脏T细胞IL-2和IFN-γ蛋白表达比Control组明显降低（37．4±2．0）VS（76．1±3．7）ng／L；（30．6±1．3）VS（49．9±2．6）ng／L（P〈0．01），IL-2和IFN-γ mRNA表达也明显降低（P〈O．01）；与Trauma组比较，T＋EA组IL-2和IFN叫蛋白表达明显增加（58．3±1．1）VS（37．4±2．0）ng／L；（39．2±1．5）vs（30．6±1．3）ng／L（P〈O．01），IL-2和IFN-γ mRNA表达也明显增加（P〈O．01）；与Control组比较，EA组的IL-2和iFN蛋白表达差异无统计学意义。与Control组比较，Trauma组p-ERKl／2、NF-KB和AP．1DNA连接活性明显降低（P〈0．01）；与Trauma组比较，T＋EA组ERKl／2、NF—KB和AP．1DNA连接活性明显增加（P〈0．01）；与Control组比较，EA组的NF—KB和AP-1DNA连接活性差异无统计学意义。结论EA“足�Objective To investigate effect of electroacupuncture （EA） on interleukin-2（IL-2）and interferon-gamma（IFN-γ） protein and mRNA expression, as well as extracellular- regulated protein kinasel/2 （ERK1/2） signaling pathway, and evaluate the signaling regulatory mechanism of EA effect. Methods Thirty-two male Sprague-Dawley （SD） rats were divided randomly into four groups （Control, EA, Trauma, and T＋EA group, n=8）. Rats were anesthetized, and aseptically incised longitudinally to a length of 6 cm along the dorsal median line and 5 cm along the abdominal median line, and the abdominal viscera were exposed for 3 min. EA was applied at bilateral ＂Zusanli＂ and ＂Lanwei＂ acupoints. At postoperative day 3, splenic T cells were isolated using the nylon column method. IL-2 and interferon-gamma（IFN-γ） protein and mRNA expressions were assayed by ELISA and reverse transcriptionpolymerase chain reaction （RT-PCR）, respectively. We examine the activity of ERK1/2 using Western-blot, and the DNA binding activity of nuclear faetor-KB （NF-KB） and activator protein-1 （AP-1） using Trans-AM ELISA-based kits. Results IL-2 and IFN-γ protein expression in splenic T cells of rats in Trauma group were markedly decreased 3 days after operation than Control group （37.4±2.0） vs （76.1±3.7） ng/L, （30.6±1.3） vs （49.9±2.6） ng/L, P〈0.01）, IL-2 and IFN-γ mRNA expression in Trauma group were also decreased （P〈O.O1）. This was accompanied with a significant depression in the activity of ERK1/2, NF-KB and AP-1 （P〈0.01, Trauma group vs Control group）. IL-2 and IFN-γ production in T＋EA group were significantly increased compared with Trauma group （58.3γ1.1） vs （37.4γ2.0） ng/L, （39.2γ1.5） vs （30.6γ1.3） ng/L, P〈O.O1）, and IL-2 and IFN-γ mRNA expression, ERK1/2 activation as well as NF-KB and AP-1 DNA binding activity were also increased （P〈O.01）. Compared with Control group, the protein and mRNA expressions of IL-2 and IFN-γ, and the act

关 键 词：电针刺 手术创伤 白细胞介素-2 γ-干扰素 细胞外信号调节激酶 

分 类 号：R245[医药卫生—针灸推拿学]