国际麻醉学与复苏杂志   2012, Issue (2): 2-2
    
电针刺对创伤大鼠脾T细胞IL-2、IFN-γ及ERK通路的影响
王坤, 王玉, 王国年1()
1.哈尔滨医科大学附属第三医院
Effects of electroacupuncture on IL-2、IFN-γ and ERK1/2 signaling pathways in the splenic T cells of traumatized rats
 全文:
摘要:

目的 观察电针刺对手术创伤大鼠脾脏T细胞IL-2、IFN-γ和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SD大鼠随机均分为:对照组(Control组)、单纯电针刺组(EA组)、创伤组(Trauma组)和创伤后电针刺组(T + EA组)。大鼠麻醉后,沿背部中线、腹部中线行6 cm和5 cm的纵向切口,暴露腹部脏器3 min,制作免疫抑制模型。选双侧“ 足三里” 和“阑尾”穴行电针刺。于术后3天无菌取脾,尼龙毛柱法分离脾T淋巴细胞,用ELISA和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ERK1/2表达;NF-κB和AP-1活性测定采用Trans-AM ELISA-based kits分析法。结果 与Control组比较,Trauma组脾脏T细胞IL-2和IFN-γ蛋白和mRNA表达明显降低(P < 0.01);与Trauma组比较,T + EA组IL-2、IFN-γ蛋白和mRNA表达明显增加(P < 0.01);与Control组比较,EA组的IL-2和IFN-γ蛋白表达无统计学差异。 与Control组比较,Trauma组p-ERK1/2、NF-κB和AP-1 DNA连接活性明显降低(P < 0.01);与Trauma组比较,T + EA组p-ERK1/2、NF-κB和AP-1 DNA连接活性明显增加(P < 0.01);与Control组比较,EA组的NF-κB和AP-1 DNA连接活性无统计学差异。 结论 电针刺“足三里”和“阑尾”穴可提高创伤大鼠脾脏T细胞IL-2、IFN-γ蛋白和mRNA表达,改善术后免疫抑制状态,这种保护作用与ERK1/2信号通路活化有关。推测ERK1/2信号通路可能是其免疫调节作用的分子机制。

关键词: 电针刺;手术创伤;IL-2;IFN-γ;ERK1/2
Abstract:

Objective To investigate effect of electroacupuncture (EA) on IL-2 and IFN-γ protein and mRNA expression, as well as ERK1/2 signaling pathway, and evaluate the signaling regulatory mechanism of EA effect. Methods Thirty-two male SD rats were divided randomly into four groups (Control, EA, Trauma, and T + EA group, n = 8/group). 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 IFN-γ protein and mRNA expressions were assayed by ELISA and RT-PCR, respectively. We examine the activity of ERK1/2 using Western-blot, and the DNA binding activity of NF-κB and AP-1 using Trans-AM ELISA-based kits. Results IL-2 and IFN-γ protein and mRNA expression in splenic T cells of traumatized rats were markedly decreased 3 days after operation (P < 0.01, Trauma group vs. Control group). This was accompanied with a significant depression in the activity of ERK1/2, NF-κB and AP-1 (P < 0.01, Trauma group vs. Control group). EA administration increased these cytokine production and mRNA expression (P < 0.01, T + EA group vs. Trauma group), and the ERK1/2 activation as well as NF-κB and AP-1 DNA binding activity (P < 0.01, T + EA group vs. Trauma group). Compared with Control group, the protein and mRNA expressions of IL-2 and IFN-γ, and the activity of ERK1/2, NF-κB and AP-1 in splenic T cells of T + EA group are not different. Conclusion EA may ameliorate impaired immune function following surgical trauma by regulating IL-2, IFN-γ protein and mRNA expression in splenic T cells, and, at least in part, involves the signaling pathways of ERK1/2 as well as NF-κB and AP-1.

Key words: Electroacupuncture; surgical trauma; IL-2; IFN-γ; ERK