摘要
消化性溃疡(PU)是一种临床上常见的慢性复发性疾病。近年来随着H2受体拮抗剂(H2-RA)及质子泵抑制剂(PPI)的应用,虽然能较快地促进溃疡近期愈合,但停药后,1年内65%～80%的病人复发,2年内几乎100%复发;幽门螺杆菌(Hp)根除疗法可使Hp相关性溃疡年复发率降至3%以下,但由于费用、副作用、病人依从性等问题的存在,最终使该病的复发率居高不下。另外还有一部分溃疡与Hp感染无关,因此抗PU的复发是目前亟待解决的问题。
大量研究资料显示:完全治愈的溃疡复发率很低,特别是1990年末,在夏威夷国际消化会议上,Tarnawski等首先提出了溃疡愈合质量(QOUH)的概念,使人们对溃疡复发的认识发生了改变,更多的学者开始重视QOUH的研究。因此,提高QOUH,成为抗溃疡复发的关键。临床研究表明,针灸治疗PU能取得与H2-RA相近似或者更高的治愈率,且无毒副作用,已经引起人们的关注。但从细胞凋亡的角度及凋亡因子与溃疡复发的影响方面的研究还很少。
目的通过观察针刺治疗实验性胃溃疡大鼠后,胃组织中溃疡面积的大小、HP含量及caspase-3、IFN-γ的表达及含量的变化,分析针刺疗法与胃溃疡组织中caspase-3及IFN-γ的含量关系,揭示针刺足三里促进胃溃疡愈合及防止复发的机制。
方法清洁级SD大鼠,90只。体重150—200克。全部为雌性。参照Okabe造模方法,以冰醋酸制备大鼠胃溃疡模型,分别在造模后3、8、14天,颈椎脱臼法处死,剪开腹腔,取出胃,沿胃大弯剪开,置于冰盘上,以冰生理盐水冲洗,测定溃疡指数(UI)后,溃疡周围组织分别用10%中性甲醛固定,置于超低温冰箱保存待测。进行HE染色及免疫组化染色,测量大鼠胃溃疡组织中caspase-3及IFN-γ的含量,应用计算机图像分析系统进行分析。结果做统计学分析。
结果
1、肉眼观察以冰醋酸制备大鼠溃疡模型,术后第3天,处死大鼠,肉眼观察,前壁黏膜溃疡形成率为100%,表现为典型的胃溃疡特征;正常组大鼠胃黏膜皱襞明显、色淡红、无充血水肿、无溃疡。
2、溃疡大鼠胃组织中caspase-3的含量变化与空白组(0.127±0.021)比,模型组(0.263±0.033)、电针组(0.222±0.027)caspase-3的含量均明显增高(P＜0.01);与模型组相比,电针组caspase-3含量均明显降低(P＜0.05)。
3、溃疡大鼠胃组织中IFN-γ的含量变化与空白组(0.321±0.022)比,模型组(0.363±0.046)、电针组(0.323±0.021)的含量均明显增高(P＜0.05);与模型组相比,电针组IFN-γ含量均明显降低(P＜0.05)。
结论电针可降低大鼠胃溃疡的UI指数,促进胃溃疡组织损伤的修复。电针可降低大鼠胃溃疡组织中caspase-3及IFN-γ的表达,并且降低胃溃疡组织中caspase-3及IFN-γ的含量,提示针刺治疗胃溃疡可能与针刺可降低胃溃疡组织中caspase-3及IFN-γ的含量,从而阻断细胞凋亡有关。
Objective
By observing the acupuncture treatment of experimental gastric ulcer in rats,the changes of the area of the ulcer, the content of HP and the caspase-3,IFN-γexpression in the gastric tissues.Analysed the relationship between acupuncture therapy in gastric tissues and the content of caspase-3 and IFN-γto probe the mechanism of needling ST36(Zusanli) in the promotion of the healing and prevention of the recurrence of gastric ulcer.
Methods
90 SD rats of Clean-level,150-200 grams.All female. Preparation of gastric ulcer model by using acetic acid.After operation,in 3,8,14 days,rats were killed by Cervical dislocation method.Then cut open the abdominal cavity,removed the stomach,cut the stomach along the greater curvature. Placed the tissues on ice_Plate,washed them by Ice-cold saline. After measured the ulcer index(UI),the surrounding tissue of the ulcer were fixed by 10%formaldehyde solution.The tissue were preserved in ultra-low temperature refrigerator for the next test.The tissue were staind though HE and immunohistochemical.Measure the content of caspase-3 and IFN-γin rat gastric tissues.Used computer image analysis system to analyse.The result did statistical analysis.
Results
1.Visual observation:In the rats' acetic acid ulcer model, killed rats in the three days after the operation.The percent of ulcers on the mucosal of the anterior wall was 100%.The performance were typical gastric ulcer.The normal rats' gastric mucosal fold Obviously,light red,no congestion and edema,no ulcer.
2.The change of the caspase-3's content in ulcer rat gastric tissue:The content of the caspase-3 in the blank group was 0.127±0.021,which in the model group was 0.263±0.033,in the electro-acupuncture group was 0.222±0.027.The caspase-3's levels of model group and electro-acupuncture group were significantly higher(P＜0.01)than the blank group's.Compared with model group,the content of caspase-3 of EA group were significantly lower(P＜0.05).
3.The change of the IFN-γ's content in ulcer rat gastric tissue:The content of the IFN-γin the blank group was 0.321±0.022,which in the model group is 0.363±0.046,in the electro-acupuncture group was 0.323±0.021.The IFN-γ's levels of model group and electro-acupuncture group were significantly higher(P＜0.05) than the blank group's.Compared with model group,the content of IFN-γof EA group were significantly lower(P＜0.05).
Conclusion
Electro-acupuncture may reduce the UI and promote the recovery of gastric ulcer.Electro-acupuncture reduces the expression of caspase-3 and IFN-γin the rats' gastric tissue.The results Suggest that acupuncture is effective in the treatment of gastric ulcer and its mechanism might be relative to its effectiveness in reducing the content of caspase-3 and IFN-γin gastric tissues.Thereby acupuncture can block apoptosis.
The reason why acupuncture can treat gastric ulcer may be associated with the effect that acupuncture can
引文
[1]Katz J.Multicenter study of Helicobacter pylori infection prevalence in patients with chronic gastro duodenal disease.Various epidemionlogic features[J].Acta Gastroenterol Latinoam,1997,27(6):253-257
[2]姒健敏,王杭勇,蔡建庭,等.中国人溃疡病流行状况及其治疗依从性[J].中华内科杂志,2000,39(3):153-155
[3]余跃,候晓华.细胞因子在幽门螺杆菌致病过程中的作用[J].国外医学·消化系疾病分册,1999,18(2):77-78.
[4]唐晖.十二指肠溃疡球部粘膜IL-18,IFN-γ水平变化及其临床意义.时珍国医国药,2000,16(5):411-412
[5]NecefliA,Tulumoglu B,G irlsM,et al.The effect of melation on TNB s-induced colitis[J].Dig D is Sci,2006,51(9):1538-1545.
[6]郑世营,葛锦锋,赵 军,等.外源性FasL基因诱导肺癌细胞caspase-3的表达及凋亡[J].肿瘤防治杂志,2004,3(11):273-277
[7]朱跃红,卢放根,刘祥.TLR、Caspase-3和Fas在胃癌、胃溃疡中的表达及意义.临床肿瘤学杂志,2007,12(1):37-40.
[8]关景明,赵瑞波,程艳秋,等.幽门螺杆菌感染与胃上皮细胞凋亡及消性溃疡愈合的关系[J].中国内镜杂志,2001,7(1):24-26.
[9]Yamamoto N,Sakagami T,Fukuda Y,et al.Influence of Helicobacter pylori infection on development of stress-induced gastric mucosal injury[J].Hepatogastroenterology,1999,46(27):1740-1745
[10]Ihan A,Tepez B,Gubina M,et al.Diminished interferon-gamma production in gastric mucosa T lymphocytes after H.phlori eradication in duodenal ulcer patientis[J].Infect Immun,1998,66(12):5964-5971
[11]Lindholm C,Quidin Terracciano L,Carena I,et al.In situ correlation of cytokine secretion and apoptosis in Helicobacter pylori-associated gastritis[J].Am J Physiol Gastrointest Libver Physiol,2002,283(2):481-488
[12]Lehmann FS,Jarbrink M,Lonroth H,et al.Local cytokine response in Helicobacter phlori-infected subjects[J].J Immnuol,1997,158(2):962-967
[13]Okabe P.silonw effect of bile folw[J].Gastroenterology,1973,64:246
[14]Guth PH,Aures D,Paulsen G.Topical aspirin plus HC1gastric lesions in the rat.Cytoprotective effect of prostaglandin,cimetidine,and probanthine[J].Gastroenterology,1979,76(1):88-93.
[15]谢莉莉,刘光谱.艾灸的治疗作用和机理研究进展[J].成都中医药大学,2000,16(5):55-57
[16]严洁,常小荣,邓元江,等.电针足阳明经穴对家兔胃黏膜损伤细胞保护作用的研究[J].针刺研究,2001,26(4):264-269
[17]陈演江,肖德生,汪凤维,等.艾灸对大鼠胃溃疡模型的影响[J].针刺研究,1992,17(2):113-116
[18]Srtober W,Kelsall B,Fuss I,et al.Reciprocal IFN-γ and Tgf-βresponses regulate the occurrence of mucosal inflammation[J].Immonolgy Today 1997,18(2):61-64
[19]Rieder G,Hatz RA,Stolte M,et al.Mrna transcript for inflammatory mediators in Hp-associated gastritis[M].Gut,1995,37(suppl 1):A81-81.
[1]李阳.针刺根除幽门螺旋杆菌感染的疗效观察[J].中国针灸,1999,(10):601.
[2]孙静,察鸿盛.针灸治疗消化性溃疡40例[J].陕西中医,2000,(3):130
[3]余幼鸣,针刺足三里穴镇上腹疼痛160例疗效观察和机理探讨[J].上海针灸杂志,1997(3):10
[4]孟立强,秦媛媛.消化性溃疡的针灸治疗[J].河南中医2000(3):30
[5]罗海鸥,唐勇,蒲艺等.不同穴位配伍针刺对消化性溃疡患者血浆内皮素的影响[J].新中医,2003,35(5):51-52
[6]郑关毅,郑师碧,韦立新.针刺对消化性溃疡患者内源性多肽的影响[J].上海针灸,2002,21(2):10-11
[7]郭永明,梁宪如,邱桐等.不同针刺手法对醋酸型胃溃疡大鼠溃疡指数及血清胃泌素水平的影响[J].天津中医学院学报,2001,20(4):27-28
[8]肖凡.针刺配合中药治疗胃、十二指肠溃疡穿孔23例[J].湖南中医学院学报,1999,(1):56.
[9]田立平,邵孔健.中西医结合治疗急性溃疡病穿孔80例[J].福建中医药,2000,(3):19.
[10]任国平.针灸治疗消化性溃疡36例[J].甘肃中医学院学报,2007,24(4):44-45
[11]江庆淇,许文波,杨丹红.艾灸对大鼠胃黏膜血流量的影响及于胃肠激素的关系[J].现代康复,2000,4(12):4-41
[12]刘波.浅谈埋线治疗胃、十二指肠溃疡[J].针灸临床杂志,1999,(8):47.
[13]刘国光.穴位埋线治疗胃、十二指肠溃疡57例[J].针灸临床杂 志,1998,(4):50.
[14]黄柳和.挑筋割脂埋线疗法治疗胃、十二指肠溃疡40例[J].中国针灸,1999,(10):599.
[15]刘敏.羊肠线穴位埋藏治疗胃、十二指肠溃疡165例疗效观察[J].中国针灸,1999,(6):335.
[16]肖波.穴位埋线治疗胃及十二指肠溃疡218例[J].上海针灸杂志,2002,21(2):14
[17]王志兴,鲁新华.穴位埋线法治疗消化性溃疡43例观察[J].中医药学刊,2001,19(5):457
[18]王莹,崔敏.穴位埋线法治疗消化性溃疡128例分析[J].医用放射技术杂志,2005,(9):65
[19]李咏梅,昊杞.穴位埋线配合益气活血法治疗消化性溃疡36例临床观察[J].湖南中医药导报,2002,8(5):25-251
[20]赵贤兴,赵文奎.穴位埋线结合中药治疗十二指肠溃疡的临床观察[J].上海针灸杂志,2000,(3):11.
[21]潘苏涌.健脾活血愈疡汤合穴注足三里治疗难治性消化性溃疡60例临床观察[J].光明中医2006,21(12):3-32
[22]张富山,孙国喜.自拟疡愈汤配合穴位注射治疗消化性溃疡临床观察[J].河南中医学院学报,2003,1(18):4-50
[23]郑兴,纪东梅.维生素B1、B6、B12穴位注射治疗胃及十二指肠溃疡病人65例临床观察[J].针灸临床杂志,1997,(1):37.
[24]范万生.电针治疗急性消化性溃疡穿孔28例[J].中国针灸,2000(5):35
[25]罗庆道,罗建明.电针背俞穴对急性胃脘痛的镶痛观察[J].针刺研究,1997(12):138
[26]余明哲,杨光正.电针足三里对十二指肠溃疡大鼠肠黏膜保护作用的影响[J].上海中医药大学学报,2001,1(4):48-49
[27]袁英,孙阿娟,谢元华等.电针“足三里”对胃黏膜损伤大鼠SOD、 MDA影响的时效关系[J].针刺研究,2005 30(1):29-30
[28]梁繁荣,朱慧民,何刚,等.代针膏穴敷治疗消化性溃疡临床研究[J].中国针灸,2001,(1):7.
[29]成志荣,傅卫洪.外治法治疗胃脘痛66例[J].中医外科杂志,2001,10(1):18-19
[30]李敏.推拿配合针灸治疗消化性溃疡[J].中医外治杂志,2004,13(3):33
[31]赖茂才.治脊手法配合胸穴指压治疗消化性溃疡43例[J].中国按摩与导引,2005,21(11):14-15
[32]李建忠,魏巍.针刺加TDP照射与中西药并用治疗消化性溃疡46例[J].中医外治杂志,2006,15(2):20-21
[33]窦逾常,李雪萍.中成药配合针灸治疗胃溃疡45例临床观察[J].吉林中医药,2006,26(11):55-56
[34]金淑英,刘志敏,王晶,等.电针足三里对胃运动胃电的抑制作用与蓝斑核的递质受体关系的研究[J].中医药学刊,2002,20(20):21
[35]陈香梅,柳 滨,刘志敏,等.中缝大核微量注射5-HT等药物对家兔胃运动胃电的影响[J].白求恩军医学院学报,2004,2(1):5-7
[36]刘志敏,陈宝忠,王东岩,等.中枢5-HT受体在电针对胃运动、胃电影响中的作用[J].上海针灸杂志,2000,19(4):41-42
[37]赵艳玲,常小荣,何军锋.电针对大鼠胃肌电的针刺效应及其传导途径作用机理探讨[J].中国医学研究与临床,2006,4(5):6-8.
[38]常小荣,严洁,刘玉群,等.电针足三里和阳陵泉穴对家兔胃胆运动及脑肠肽的影响[J].世界华人消化杂志,2006,14(6):17
[39]易受乡,严洁.针刺胃经穴对M受体阻断大鼠胃电及脑肠肽的影响[J].中国中西医结合消化杂志,2002,10(4):203-205
[40]李江山,严 洁,常小荣.电针足阳明经穴对家兔胃运动和胃动素的影响[J].中国中医基础医学杂志,2004,10(3):36-39
[41]姚筱梅,姚树坤,张瑞星,等.针刺对功能性消化不良患者近端胃 动力的影响[J].世界华人消化杂志,2006,14(21):2139-2141
[42]吴学飞,郑俊江,郭永明,等.温通针法预防大鼠应激性胃黏膜损伤的实验研究[J].中国针灸,2001,21(10):609
[43]郭永明,梁宪如,吴学飞,等.醋酸型胃溃疡大鼠PGE2变化及不同针刺手法的调节效应[J].辽宁中医杂志,2002,29(6):31.
[44]郭永明,梁宪如,吴学飞,等.热补针法对胃溃疡大鼠胃窦黏液细胞超微结构的影响[J].中国针灸,2002,22(11):753
[45]黄 艾,阳仁达,严 洁,等.电针对应激性胃黏膜损伤大鼠UI、GMBF及胃黏膜形态学的影响[J].中国中医基础医学杂志,2006,12(8):637-639
[46]黎喜平,严洁,易受乡,等.电针对应激大鼠胃黏膜肠三叶因子表达及形态学的影响[J].湖南中医学院学报,2006,26(3):51-53
[47]黄裕新,赵保民.针刺对胃分泌机能的调控作用[J].针刺研究,2000,25(2):89-92
[48]王瑞辉,殷克敬,赵宗辽.电磁极针对大鼠实验性胃溃疡的治疗作用及机理研究[J].陕西中医,2004,25(1):87-89.
[49]高希言.针灸对胃黏膜屏障功能影响的实验研究[J].中国中医药科技,2001,8(5):277
[50]许冠荪,裴文芬,王月芳.艾灸对胃黏膜损伤的保护作用及其与一氧化氮的关系[J].针刺研究,1997,(3):175.
[51]张燕,于浩.针刺耳胃穴对大鼠乙酸实验性胃溃疡的保护作用及机理探讨[J].贵阳中医学院学报,1998,(2):33.
[52]刘涌.电针足三里对应激性胃黏膜损伤的保护作用[J].安徽中医学院学报,2000,(2):227.
[53]秦明,黄裕新.针刺对胃黏膜保护作用及其机制的研究进展[J].世界华人消化杂志2000,8(4):456-457.
[54]黄国峰,汤得安.电针“梁门”穴治疗大鼠实验性胃溃疡的机理探讨[J].针刺研究,1999,(1):51.
[55]黎喜平,严洁.针灸对胃黏膜损伤保护作用机制的研究进展[J].针刺研究,2005,30(1):60-63.
[56]朱舜丽,许冠荪,陈金珠,等.艾灸和针刺对实验性胃溃疡大鼠NO 含量的变化及L-NNA抑制作用的影响[J].针刺研究1997,(3):177.