电针阳陵泉在内的下合穴对急性胆囊炎模型豚鼠核因子-κB及白介素-1β的影响

Effect of electroacupuncture at Lower He-Sea points including Yanglingquan(GB 34)on nuclear factor-κB and interleukin-1βin guinea pigs with acute cholecystitis

目的:观察电针急性胆囊炎(AC)模型豚鼠下合穴对血清及胆囊组织白介素-1β(IL-1β)和胆囊组织核因子-κB(NF-κB)表达的影响,探讨胆的下合穴阳陵泉对胆腑病变是否存在相对特异性.方法:将82只健康豚鼠按照随机数字表法随机分为空白组12只,模型组、阳陵泉组、足三里组、上巨虚组和下巨虚组,每组14只,均雌雄各半.除空白组外,其余组豚鼠均予胆囊内注射大肠杆菌建立AC模型.空白组正常饲养而无特殊处理;模型组每日捆绑30 min但不做电针治疗;接受电针治疗的4组每天捆绑后针刺相应下合穴,并使用SDZ-V型电子针疗仪进行刺激.造模成功并治疗5 d后,行豚鼠腹主动脉采血,取胆囊组织进行苏木素-伊红(HE)染色,观察各组豚鼠胆囊形态学变化,酶联免疫吸附(ELISA)法检测血清中IL-1β含量,免疫组织化学(IHC)法检测胆囊NF-κB及IL-1β的表达.结果:造模后第3天,接受造模的5组豚鼠精神萎靡,食欲下降,活动明显减少;弓背倦怠,皮毛松,无光泽,大便稀;每组各取2只(雌雄各1只,不纳入统计),处死后取整个胆囊,肉眼观察胆囊,胆囊壁不同程度增厚,胆汁颜色呈深绿色,不透明,有颗粒悬浮或堆积;光镜下观察,胆囊黏膜下血管充血,有黏膜水肿、溃疡、坏死、脱落,固有层有大量炎细胞浸润,提示豚鼠AC造模成功.与模型组比,接受电针治疗的4组胆囊组织损伤均有不同程度修复,血清中IL-1β含量均降低(P<0.01或P<0.05),胆囊组织中IL-1β含量均降低(P<0.01或P<0.05).与模型组比较,阳陵泉组NF-κB表达明显降低(P<0.01),足三里组、上巨虚组和下巨虚组NF-κB表达无统计学差异(均P>0.05).与阳陵泉组比,足三里组、上巨虚组和下巨虚组胆囊组织损伤较严重,血清中IL-1β含量及胆囊组织中NF-κB及IL-1β表达增高(P<0.01或P<0.05).结论:电针胃、大肠、小肠及胆的下合穴均可对AC模型豚鼠产生疗效,减轻炎性症状.阳陵泉对AC模型豚鼠的干预效应优于足三里、上巨虚和下巨虚.电针阳陵泉治疗AC的机制可能是通过调节IL-1β及NF-κB等调控炎症反应,改善胆囊组织损伤.

Objective:To observe the effect of electroacupuncture(EA)at Lower He-Sea points on the expression levels of interleukin-1β(IL-1β)in the serum and gallbladder tissues,and nuclear factor-κB(NF-κB)in gallbladder tissues of the guinea pigs with acute cholecystitis(AC),and to explore whether Yanglingquan(GB 34),the Lower He-Sea point pertaining to Dan Fu(gallbladder),is relatively specific for the Dan Fu(gallbladder)disorders.Methods:Eighty-two healthy guinea pigs were randomly divided into 6 groups according to the random number table method,a blank group,a model group,a Yanglingquan(GB 34)group,a Zusanli(ST 36)group,a Shangjuxu(ST 37)group,and a Xiajuxu(ST 39)group,with 12 guinea pigs in the blank group while 14 in the other groups,respectively,half males and half females in each group.Except for the blank group,guinea pigs in the other groups were injected with E.coli into the gallbladder to establish AC models.Guinea pigs in the blank group were fed routinely without special treatment;those in the model group were daily tied up for 30 min without EA treatment;those in the 4 groups receiving EA treatment were acupunctured at the corresponding Lower He-Sea points after daily binding and stimulated with the SDZ-V EA instrument.After successful modeling and treatment for 5 d,blood was collected from the abdominal aorta of the guinea pigs,and the gallbladder tissues in each group were isolated for hematoxylin-eosin(HE)staining to observe the morphological changes.Enzyme-linked immunosorbent assay(ELISA)was used to detect the serum IL-1βlevel,and immunohistochemistry(IHC)was used to detect the expression levels of NF-κB and IL-1βin gallbladder.Results:On the 3rd day after modeling,the guinea pigs in the five groups with modeling were mentally depressed with decreased appetite,significantly reduced activities,slouch,lassitude,slack and matted fur,and loose stools;two guinea pigs were selected from each group(one male and one female,not included in the final statistics)to isolate the gallbladder after sacrifice;macroscopic observation showed that the gallbladder wall was differently thickened;the bile color was dark green and opaque with particles suspended or accumulated;light microscope observation showed that the submucosal blood vessels of the gallbladder were congested,along with mucosal edema,ulceration,necrosis,shedding,and a large number of inflammatory cells infiltrating in the lamina propria,indicating that the AC model was successfully prepared.Compared with the model group,the gallbladder tissue injuries of the four groups receiving EA treatment were all differently repaired,the serum IL-1βlevels were reduced(P<0.01 or P<0.05),and the IL-1βlevels in the gallbladder tissues were reduced(P<0.01 or P<0.05).Compared with the model group,the NF-κB expression level in the Yanglingquan(GB 34)group was significantly reduced(P<0.01),but was not statistical different in the Zusanli(ST 36)group,Shangjuxu(ST 37)group and Xiajuxu(ST 39)group(all P>0.05).Compared with the Yanglingquan(GB 34)group,the gallbladder tissues of the Zusanli(ST 36)group,Shangjuxu(ST 37)group and Xiajuxu(ST 39)group were more severely damaged,and the expression levels of serum IL-1β,the NF-κB and IL-1βin the gallbladder tissues were increased(P<0.01 or P<0.05).Intervention effect of Yanglingquan(GB 34)on AC guinea pigs was superior to that of Zusanli(ST 36),Shangjuxu(ST 37)and Xiajuxu(ST 39).Conclusion:EA at the Lower He-Sea points of the stomach,large intestine,small intestine and gallbladder can produce curative effects on AC guinea pigs and reduced the inflammatory symptoms.Intervention effect of Yanglingquan(GB 34)on AC guinea pigs is superior to that of Zusanli(ST 36),Shangjuxu(ST 37)and Xiajuxu(ST 39).The mechanism of EA at Yanglingquan(GB 34)in treating AC may be regulating IL-1βand NF-κB to control the inflammatory response and improve the gallbladder tissue damage.