【作者】 莫莉；

【导师】 皮明钧；

【作者基本信息】 湖南中医药大学 ， 中医基础理论， 2005， 硕士

【摘要】 目的:探讨半夏泻心汤及其拆方对幽门螺杆菌感染小鼠胃粘膜形态学的影响及其作用机理,并比较半夏泻心汤全方及各拆方组对幽门螺杆菌感染小鼠胃组织的修复作用,为临床防治幽门螺杆菌感染性胃病提供实验依据。方法:将120只C₅₇BL小鼠随机分为10组:Ⅰ半夏泻心汤全方组,Ⅱ半夏组,Ⅲ甘温组,Ⅳ苦寒组,Ⅴ半夏加苦寒组,Ⅵ半夏加甘温组,Ⅶ苦寒加甘温组,Ⅷ阳性药物对照组,Ⅸ模型纽,Ⅹ空白组。每组各12只小鼠,雌雄各半。先连续4天用氨苄西林给小鼠清肠消毒,第5天开始造模:第Ⅰ组至第Ⅸ组每只小鼠灌胃幽门螺杆菌菌液0.2ml,灌胃前禁食禁水,每隔一天灌胃一次,共8次。末次灌胃造模两天后,随机抽取四只小鼠进行胃粘膜细菌培养和尿素酶试验。结果:细菌培养,革兰氏染色镜检发现G^-弧形杆菌,菌尿素酶试验阳性,证实造模成功后开始药物灌胃。第Ⅰ组至第Ⅶ组按每只小鼠体重给予等效剂量的中药灌胃;第Ⅷ组灌胃等效剂量的氨苄青霉素,用蒸馏水稀释成所需浓度。模型组和空白组每只小鼠灌胃0.2ml的蒸馏水。连续灌胃8天,每天1次,灌药前禁水禁食6小时。灌胃结束后,动物禁食不禁水12小时,次日心脏采血处死动物,并完整取胃,观察胃组织的病理形态改变和检测胃细胞凋亡及T淋巴细胞亚群CD4⁺和CD8⁺的表达。结果:1.半夏泻心汤全方组、阳性药物对照组大多数未见明显的炎细胞浸润,此两组与空白组相比无显著性差异（P＞0.05）。苦寒加甘温组、苦寒组和苦寒加半夏组可见少量炎细胞浸润,多累及胃粘膜的1/3。模型组可见大量炎细胞浸润,累及胃粘膜的2/3甚至全层。治疗组中全方组、阳性药物对照组、苦寒加甘温组与模型组对比均有显著性差异（P＜0.01）。2.模型组凋亡细胞增多,电镜下可观察到凋亡小体。药物治疗组中全方组、阳性药物对照组、苦寒加甘温组、苦寒组和苦寒加半夏组与模型组相比,均能有效减少胃粘膜细胞凋亡,比较有显著性差异（P＜0.01）。3.模型组CD4⁺明显降低,CD8⁺明显增多,CD4⁺/CD8⁺比例下降。各组CD4⁺的表达依次为阳性药物对照组＞全方组＞苦寒加甘温组＞苦寒组＞半夏加苦寒组＞半夏加甘温组＞甘温组＞半夏组＞模型组。各组CD8⁺表达水平由高至低依次为模型组＞半夏组＞半夏加甘温组＞甘温组＞苦寒组＞半夏加苦寒组＞苦寒加甘温组＞全方组＞阳性药对照组＞空白组药物治疗组。其中全方组、阳性药物对照组、苦寒加甘温组、苦寒组和苦寒加半夏组在胃粘膜细胞CD4⁺和CD8⁺的表达水平上以及CD4⁺/CD8⁺比例都与模型组有显著性差异（P＜0.01）。结论:1.幽门螺杆菌感染可导致胃粘膜受损、细胞凋亡增多和T细胞亚群失调。抑制幽门螺杆菌活性,祛除邪气是治疗幽门螺杆菌感染胃粘膜相关疾病的重要环节。2.半夏泻心汤对于幽门螺杆菌感染小鼠胃粘膜有较好的修复作用。主要体现在半夏泻心汤保护胃粘膜,抑制细胞凋亡和调节CD4⁺与CD8⁺的表达。3.半夏泻心汤全方组与各拆方组比较,全方组对幽门螺杆菌感染的修复作用明显优于各拆方组,其原因与半夏泻心汤寒热并用、辛开苦降、扶正祛邪的精当配伍密切相关。各拆方组中以扶正祛邪兼施的苦寒加甘温组疗效最佳。综合各指标检测结果,各药物组对幽门螺杆菌感染胃粘膜的修复作用从大至小依次为阳性药物对照组＞半夏泻心汤全方组＞苦寒加甘温组＞苦寒组＞半夏加苦寒组＞甘温组＞半夏加甘温组＞半夏组。更多还原

【Abstract】 AIM:Probe into tuber of Pinellia Heart-Purging Decoction and dismantle square to infect little mouse stomach influence and function mechanism,mucous membrane of morphology to pylorus spiral shell bacillus,and the whole side dismantle square group infect little mouse repair function that stomach organizes to pylorus spiral bacillus each, offer the experiment basis for the fact that prevent and cure the infectious stomach trouble of pylorus spiral bacillus clinically。Method:Divide the little mouse of 120 C57BLs into 10 groups at random:ⅠPinellia Heart-Purging Decoction all square group,Ⅱtuber of pinellia group,Ⅲsweet warm group,Ⅳbitter cold group,Ⅴtuber of pinellia add bitter cold group,Ⅵtuber of pinellia add sweet warm group,Ⅶbitter cold add sweet warm group,Ⅷpositive medicine contrast group,Ⅸmodel group,Ⅹnormal group.12 little mice respectively of every group,the male and female are half and half.Give the little mouse clear intestines to disinfect with the ammonia and benzyl and west forest in 4 days in succession first,begin to make the mould on the 5th day:Ⅰgroup irritate stomach pylorus spiral shell bacillus fungus liquid 0.2ml toⅨgroup each little mouse,irritate stomach fasting prohibitted water, irritate the stomach once every other day,amounts to 8 times.The end time irritate stomach build one day such as mould,collect four little mouse carry on stomach mucous membrane bacterium train with the urea enzyme testing at random.Result:Bacterium train,leather blue one dye mirror is it find G-curved bacillus to examine,fungus urea enzyme tests the masculine gender,is it make mould begin medicine irritate the stomach after succeeding to verify.Ⅰgroup offer equivalent traditional Chinese medicine of dosage irritate the stomach toⅦgroup according to each little mouse weight;Ⅷgroup irritate stomach equivalent ammonia benzyl penicillin of dosage,dilute with the distilled water into the necessary density.Model group and normal group each little mouse irritate distilled water,stomach of 0.2ml.Irritate the stomach for 8 days in succession,1 times a day,prohibit water fasting for 6 hours before irritating medicines.Irritate after the stomach,animal fasting can’t help 12 hours such as ink,the heart collects blood and puts animals to death next day,it is and the intact it fetch stomach,it observe pathology shape where stomach organize change and measure stomach the cells not to wither dying and T lymphocyte groups of CD4~+and CD8~+expression.Result:1.Pinellia Heart-Purging Decoction square group,positive medicine contrast most of groups meet the obvious inflammation cell soak into all,compared with normal group this two groups do not have a significance difference(P＞0.05).Bitter cold add sweet warm group, bitter cold group and bitter cold add tuber of pinellia group can see a small amount of inflammation cell soak into,involve 1/3 of the stomach mucous membrane more.Model group can see a large number of inflammation cell soak into,involve 2/3,stomach of mucous membrane even the whole layer.Treat group square group,positive medicine contrast group,bitter cold add sweet warm group and model group is it have significance difference to compare with all(P＜0.01).2.Model group is it die cell increase,can is it is it lose the little body to wither to observe under the electric mirror to wither.Medication group square group,positive medicine contrast group,bitter cold add sweet warm group,bitter cold group and bitter cold add tuber of pinellia group compare with model group all,can reduce the stomach mucous membrane cell and wither and die,relatively has a significance difference(P＜0.01)effectively.3.The model group CD4~+is obviously reducing,CD8~+not obviously it increase,CD4~+/drop CD8~+ the proportion.Every group CD4 ~+expression to last group＞All square groups＞The bitter cold adds the sweet warm group＞Bitter cold group＞ The tuber of pinellia adds the bitter cold group＞The tuber of pinellia adds the sweet warm group＞Sweet warm group＞Tuber of pinellia group＞Model group.Every group CD8 express competence from high to successively a model group＞Tuber of pinellia group＞The tuber of pinellia adds the sweet warm group＞Sweet warm group＞Bitter cold group＞The tuber of pinellia adds the bitter cold group＞The bitter cold adds the sweet warm group＞All square groups＞The positive medicine contrasting and organizing＞The normal group medicine treats the group. All square groups among them,the positive medicine is contrasted and organized,the bitter cold adds the sweet warm group,last group at stomach mucous membrane cell CD4 ~+ the and CD8 ~+bitter cold group and bitter cold expression competence at and CD4 ~+/CD8~+ the proportion have significance difference as model group(P＜0.01).Conclusion:1.Pylorus spiral shell bacillus is it can cause stomach to be mucous membrane damaged to infect,cell is it is it increase with T cell inferior group lacking proper care to die to wither.Suppressing the pylorus spiral shell’s bacillus activation,it is to treat the important link of infecting the relevant diseases of mucous membrane of stomach of spiral shell’s bacillus of pylorus to dispel perverse trend.2.Pinellia Heart-Purging Decoction infect to pylorus spiral shell bacillus little mouse stomach mucous membrane have better repair function.Mainly it embody in it rush down tuber of pinellia there aren’t heart the soups,suppress cell not to wither not dying and not regulating it CD4 ~+and CD8 ~+ expression.3.Pinellia Heart-Purging Decoction all square group with dismantle square group compare,square group all obviously superior to each dismantling the square group to pylorus spiral shell repair function that bacillus is infect each,rush down heart soup fever and chills of reason of it and tuber of pinellia and with,the more hard to turn on more bitter to lower,compatibility not precise and appropriate not strengthen body resistance and eliminating evil closely related with. Dismantle square group in order to strengthen body resistance and eliminating evil each and concurrently bitter cold that construct add sweet warm curative effect best group.Synthesize every index measure result, every medicine group infect to pylorus spiral shell bacillus repair function,stomach of mucous membrane from heavy to light to contrast group＞Pinellia Heart-Purging Decoction in the all square groups of heart soup＞The bitter cold adds the sweet warm group＞Bitter cold group＞The tuber of pinellia adds the bitter cold group＞Sweet warm group＞The tuber of pinellia adds the sweet warm group＞Tuber of pinellia group.更多还原