艾灸对幽门螺杆菌胃炎大鼠血清免疫学作用研究  

Serumimmunological study of moxibustion on helicobacter pylori gastritis in rats

作　　者：彭艳[1] 易受乡[1] 封迎帅[2] 史冬梅[1] 侯艳玲[1] 林亚平[1] 

机构地区：[1]湖南中医药大学针灸推拿学院经穴与脏腑相关重点研究室,国家中医药管理局三级实验室,湖南长沙410007 [2]湖南省人民医院

出　　处：《中国针灸》2014年第8期783-790,共8页Chinese Acupuncture & Moxibustion

基　　金：国家自然科学基金资助项目:81072867;湖南省高校创新平台开放基金项目:12K088

摘　　要：目的:探讨艾灸保护胃黏膜损伤的免疫学机制。方法:将40只健康SD大鼠随机分为4组,即空白组、模型组、艾灸穴位组(模型+艾灸穴位)、艾灸非穴组(模型+艾灸穴位对照点),每组10只。艾灸穴位组和艾灸非穴组于造模前8天即予艾灸干预,穴取"足三里""中脘""关元""脾俞""胃俞"穴或以上穴旁开对照点,模型组仅捆绑不治疗,均每日1次,连续16d。采用幽门螺杆菌(Hp)灌胃建立大鼠Hp胃炎模型,观察大鼠胃黏膜苏木精-伊红(HE)染色镜检炎性反应程度,酶联免疫法(Elisa)检测各组大鼠血清热休克蛋白72(HSP72)及胃组织肿瘤坏死因子α(TNF-α)和白细胞介素1β(IL-1β)含量,实时定量聚合酶链式反应(PCR)法检测外周血单核细胞Toll样受体(TLR2mRNA、TLR4mRNA、CD14mRNA)、髓样分化因子88(MyD88mRNA)的表达,蛋白免疫印迹(Western Blot)法检测外周血单核细胞NFκB核因子κB、IκBα核因子κB的抑制蛋白的含量。结果:与空白组比较,模型组大鼠胃黏膜涂片革兰氏染色检测出Hp,胃黏膜组织HE染色镜检炎性反应程度评分积分值增高,血清HSP72含量及胃组织TNF-α、IL-1β含量明显升高,单核细胞TLR2mRNA、4mRNA、CD14mRNA、MyD88mRNA、NFκB表达增加(P<0.01),IκBα表达降低(P<0.05);经艾灸"足三里"等穴预处理者,艾灸穴位组大鼠胃黏膜组织HE染色镜检炎症程度积分值减低,血清HSP72含量升高,胃组织TNF-α、IL-1β含量及单核细胞TLR2、4mRNA、CD14mRNA、MyD88mRNA、NFκB表达降低(P<0.01),IκBα表达升高(P<0.05);艾灸非穴组与模型组比较差异无统计学意义(P>0.05)。结论:艾灸穴位预处理可诱导血清HSP72的高表达,通过与TLR2、4受体结合启动受体后信号转导途径,调控下游信号物质的释放,从而调节机体相关免疫物质的释放,减轻Hp胃炎大鼠胃黏膜损伤。Objective To explore the immune mechanism of moxibustion on protecting gastric mucosa injury. Methods Forty healthy SD rats were randomly divided into four groups： a blank group, a model group, a moxi- bustion acupoint group and a moxibustion non-acupoint group, 10 rats in each one. Eight days before model estab- lishment, moxibustion at ＂Zusanli＂ （ST 36）, ＂Zhongwan＂ （CV 12）, ＂Guanyuan＂ （CV 4）, ＂Pishu＂ （BL 20） and ＂Weishu＂ （BL 21） was applied in the moxibustion acupoint group while these acupoints＇ controlled points were se- lected in the moxibustion non-acupoint group, and no treatment was given in the model group, once a day in three groups for continuous 16 days. The helicobacter pylori （Hp） model was established by intragastric administration of Hp. HE staining microscopic examination was used to observe inflammation severity in gastric mucosa, and enzyme-linked immunosorbent assay （ELISA） was adapted to measure content of heat shock protein （HSP） 72, TNF-a and IL- lβ, and real-time quantitative PCR was used to measure the expression of TLR2 mRNA, TLR4 mRNA, CD14 mRNA and MyD88 mRNA in peripheral blood mononuclear cells, and western blot method was used to measure contentof NF^B and IκBa in peripheral blood mononuclear cells. Results Compared with the blank group, the expression of HP could be seen in the smear of gastric mucosa by Gram＇s staining in the model group; the inflammation severi- ty score was obviously increased as well as content of serum HSP 72 and TNF-α and IL-113 in gastric tissue; and expression of TLR2, 4 mRNA, CD14 mRNA, MyD88 mRNA, NFκB was increased （P〈0.01）, but the expres- sion of IκBa was reduced （P〈0.05）. After the moxibustion, the inflammation severity score was reduced in the moxibustion acupoint group, and the content of serum HSP 72 was increased, and the expression of TNF-a and IL- 1β in gastric tissue and expression of TLR2 mRNA＇ TLR4 mRNA, CD14 mRNA, MyD88 mRNA and NF^B were reduced （P^0.01）, but

关 键 词：胃炎大鼠 幽门螺旋杆菌 艾灸 穴位 热休克蛋白72(HSP72) TOLL受体 髓样分化因子88 

分 类 号：R245.8[医药卫生—针灸推拿学]