胃饥饿素介导幽门螺杆菌相关消化不良的动物实验研究

doi:10.3877/cma.j.issn.1674-1358.2022.05.005

目的

建立慢性幽门螺杆菌感染的动物模型，在此模型中通过加强胃饥饿素的表达，探讨胃饥饿素在幽门螺杆菌相关性消化不良中的影响。

方法

选取4周龄雌性C57BL/6小鼠24只，随机分为对照组（蒸馏水灌胃+生理盐水腹腔注射）、实验组A（幽门螺杆菌菌液灌胃+生理盐水腹腔注射）和实验组B（幽门螺杆菌菌液灌胃+雷帕霉素腹腔注射）。实验结束后处死小鼠，分别检测3组小鼠胃饥饿素表达水平、胃内容物残留率及小肠推进率及HE染色观察小鼠胃炎的病理变化。采用独立样本t检验分析小鼠的胃排空率、小肠推进速度、胃饥饿素分泌水平；多组间数据的整体比较采用ANOVA检验，再应用LSD-t进行组间两两比较。

结果

实验组A小鼠胃残留率[（43.6 ± 7.2）%]较对照组小鼠[（61.6 ± 3.8）%]显著下降，差异有统计学意义（t = 4.579、P = 0.013）；实验组B小鼠胃残留率[（61.6 ± 3.8）%]较实验组A[（43.6 ± 7.2）%]升高，差异均有统计学意义（t = 5.232、P = 0.012）。对照组、实验组A、实验组B小鼠的小肠推进率经两两比较差异均无统计学意义（P均> 0.05）。与对照组小鼠的胃饥饿素[（39.6 ± 1.3）%]分泌相比，实验组A小鼠胃饥饿素[（25.4 ± 5.0）%]分泌显著减少（t = 6.104、P < 0.001），且病理染色发现伴有胃炎发生；实验组B小鼠胃饥饿素[（31.8 ± 4.5）%]较实验组A小鼠分泌显著增多，差异有统计学意义（t = 3.196、P = 0.041），同时病理染色提示胃炎得到不同程度改善。

结论

提高胃饥饿素水平可促进幽门螺杆菌相关消化不良症状的改善。

关键词: 幽门螺杆菌, 消化不良, 胃排空, 胃饥饿素, 雷帕霉素

Objective

To establish an animal model of chronic Helicobacter pylori (H. pylori) infection, and to investigate the effect of ghrelin on H. pylori related dyspepsia by increasing ghrelin expression.

Methods

Twenty-four 4-week-old female C57BL/6 mice were randomly divided into control group (water gavage + saline injection), group A (H. pylori gavage + saline injection) and group B (H. pylori gavage + rapamycin injection). The mice were sacrificed at the end of the experiment, and the expression level of ghrelin, residual rate of gastric contents and small intestine propelling rate of the three groups were detected, and the pathological changes of gastritis in mice were observed by HE staining. Independent sample t test was used to analyze the gastric emptying mice, intestinal propulsion speed and ghrelin secretion of mice. Overall comparison of data between multiple groups was performed by ANOVA test and pairwise comparisons between the groups were performed by LSD-t test.

Results

The gastric residual rate of group A mice [(43.6 ± 7.2)%] was significantly lower than that of control mice [(61.6 ± 3.8)%], with significant difference (t = 4.579, P = 0.013). The gastric residual rate of group B mice [(61.6 ± 3.8)%] was higher than that of group A mice (43.6 ± 7.2%), with significant difference (t = 5.232, P = 0.012). There was no significant difference in small intestine propulsion mice between two groups (all P > 0.05). Compared with ghrelin secretion of control mice [(39.6 ± 1.3)%], ghrelin secretion of group A mice significantly reduced [(25.4 ± 5.0)%], (t = 6.104, P < 0.001), accompanied by gastritis. Compared with ghrelin secretion of group A mice, group B mice secretion ghrelin [(31.8 ± 4.5)%] was significantly increased (t = 3.196, P = 0.041), and the levels of gastritis were improved.

Conclusions

Increased ghrelin levels could alleviate symptoms of H. pylori-related dyspepsia.

Key words: Helicobacter pylori, Dyspepsia, Gastric emptying, Ghrelin, Rapamycin

图1 使用产气袋培养幽门螺杆菌模式图

图2 幽门螺杆菌固体培养图注：幽门螺杆菌菌落颜色透明色泽很浅为半透明色，在强光下观察结果不明显。因此使用侧位观察，可见透明大量透明针尖样菌落铺满平板

图3 幽门螺杆菌液体培养显微镜下图注：幽门螺杆菌液体培养后未染色直接于电子显微镜下图片，可见图中存在大量形态各异的棒状细菌，呈海鸥形，弯曲状，直棒状等多种形态

表1 三组小鼠胃残留率和小肠推进率（± s，%）

组别	只数	胃残留率	小肠推进率
对照组	6	61.6 ± 3.8	55.9 ± 3.9
实验组A	6	43.6 ± 7.2	53.4 ± 6.8
实验组B	6	61.2 ± 6.6	62.4 ± 4.4
F值		14.658	2.394
P值		< 0.001	0.128
^at值		4.579	0.646
^aP值		0.013	0.539
^bt值		5.232	0.754
^bP值		0.012	0.468
^ct值		0.114	2.411
^cP值		0.912	0.037

表1 三组小鼠胃残留率和小肠推进率（± s，%）

组别	只数	胃残留率	小肠推进率
对照组	6	61.6 ± 3.8	55.9 ± 3.9
实验组A	6	43.6 ± 7.2	53.4 ± 6.8
实验组B	6	61.2 ± 6.6	62.4 ± 4.4
F值		14.658	2.394
P值		< 0.001	0.128
^at值		4.579	0.646
^aP值		0.013	0.539
^bt值		5.232	0.754
^bP值		0.012	0.468
^ct值		0.114	2.411
^cP值		0.912	0.037

图4 小鼠胃肠蠕动功能模式解剖图注：A、B、C分别为3组小鼠灌胃结扎后胃组织的图像，可见A、C中的胃内仍留存有较多的半固体营养液，B中的胃内半固体营养液含量明显减少；D为小鼠灌胃后游离小肠的图像

图5 幽门螺杆菌鉴定与小鼠胃组织染色图注：A：对照组（左）与实验组（右）的小鼠胃组织尿素酶实验结果对比；B和C中所指为Hp在胃黏膜中定植；E、F、G分别为对照组、实验组A和实验组B胃组织

表2 ELISA法检测3组小鼠血清胃饥饿素水平（± s，pg/ml）

组别	数量	血清胃饥饿素
对照组	6	39.6 ± 1.3
实验组A	6	25.4 ± 5.0
实验组B	6	31.8 ± 4.5
F值		19.222
P值		< 0.001
^at值		6.104
^aP值		< 0.001
^bt值		3.196
^bP值		0.041
^ct值		4.018
^cP值		0.002

表2 ELISA法检测3组小鼠血清胃饥饿素水平（± s，pg/ml）

组别	数量	血清胃饥饿素
对照组	6	39.6 ± 1.3
实验组A	6	25.4 ± 5.0
实验组B	6	31.8 ± 4.5
F值		19.222
P值		< 0.001
^at值		6.104
^aP值		< 0.001
^bt值		3.196
^bP值		0.041
^ct值		4.018
^cP值		0.002

[1]	Stanghellini V, Chan FK, Hasler WL, et al. Gastroduodenal disorders[J]. Gastroenterology,2016,150(6):1380-1392.
[2]	刘文忠. 幽门螺杆菌感染,慢性胃炎和功能性消化不良[J]. 中华消化杂志,2002,22(10):1-2.
[3]	史萍慧，刘丽.加味柴胡疏肝散联合幽门螺杆菌根除方案治疗幽门螺杆菌感染相关慢性胃炎的临床研究[J/CD]. 中华实验和临床感染病杂志(电子版),2017,11(5):496-499.
[4]	Sugano K, Tack J, Kuipers E J, et al. Kyoto global consensus report on Helicobacter pylori gastritis [J]. Gut,2015,64(9):1353-1367.
[5]	Koletzko L, Macke L, Schulz C, et al. Helicobacter pylori eradication in dyspepsia: New evidence for symptomatic benefit[J]. Best Pract Res Clin Gastroenterol,2019,40-41:101637.
[6]	Calam J. Helicobacter pylori, acid and gastrin[J]. Eur J Gastroenterol Hepatol,1995,7(4):310-317.
[7]	Osawa H. Ghrelin and Helicobacter pylori infection[J]. World J Gastroenterol,2008,14(41):6327-6333.
[8]	Yanagi S, Sato T, Kangawa K, et al. The homeostatic force of ghrelin[J]. Cell Metabolism,2018,27(4):786-804.
[9]	Feng GJ, Chen Y, Li K. Helicobacter pylori promote inflammation and host defense through the cagA-dependent activation of mTORC1 [J]. J Cell Physiol,2020,235(12):10094-108.
[10]	Pena-Leon V, Perez-Lois R, Seoane LM. mTOR pathway is involved in energy homeostasis regulation as a part of the gut-brain axis[J]. Int J Mol Sci,2020,21(16):5715.
[11]	Hong X, Zhang H, Liang H, et al. Exendin-4 decreases ghrelin levels through mTOR signaling[J]. Mol Cell Endocrinol,2016,437:201-212.
[12]	Tang L, Zeng Y, Li L, et al. Electroacupuncture upregulated ghrelin in rats with functional dyspepsia via AMPK/TSC2/Rheb-mediated mTOR inhibition[J]. Dig Dis Sci,2020,65(6):1689-1699.
[13]	李梦俊，张晓荣，王婧, 等. 幽门螺杆菌感染介导IL-1β升高导致C57BL小鼠骨骼肌衰减[J]. 中国免疫学杂志,2021,37(17):2065-2069.
[14]	苏丹，唐飞，张善明, 等. 不同嫩度青砖茶改善小鼠胃肠道功能研究[J]. 湖北农业科学,2021,60(14):103-108.
[15]	Kim SE, Memon A, Kim BY, et al. Gastroprotective effect of phytoncide extract from Pinus koraiensis pinecone in Helicobacter pylori infection [J]. Sci Rep,2020,10(1):9547.
[16]	罗欣，王玉珍. 功能性消化不良发病机制研究进展[J]. 世界最新医学信息文摘,2019,19(76):91-92, 107.
[17]	柳红良，白宇宁. 基于罗马Ⅳ标准探讨功能性消化不良中西医心身诊疗思路[J]. 中国临床医生杂志,2021,49(10):1138-1141.
[18]	谢琼，卢月月，易宏锋. 上消化道症状与幽门螺杆菌感染及胃癌前疾病的相关性[J/CD]. 中华实验和临床感染病杂志(电子版),2016,10(3):319-322.
[19]	Liu B, Dong J, Wang S, et al. Helicobacter pylori causes delayed gastric emptying by decreasing interstitial cells of Cajal[J]. Exp Ther Med,2021,22(1):663.
[20]	Duval-Araujo I, Queiroz DMM, Magnago AGP, et al. Increased gastric emptying induced by Helicobacter heilmannii type 1 infection in rats[J]. J Med Microbiol,2000,49(7):627-634.
[21]	Saito Y, Suzuki H, Tsugawa H, et al. Dysfunctional gastric emptying with down-regulation of muscle-specific microRNAs in Helicobacter pylori-infected mice[J]. Gastroenterology,2011,140(1):189-198.
[22]	Verdu EF, Bercik P, Huang XX, et al. The role of luminal factors in the recovery of gastric function and behavioral changes after chronic Helicobacter pylori infection[J]. Am J Physiol Gastrointest Liver Physiol,2008,295(4):G664-G670.
[23]	Russo F, Chimienti G, Clemente C, et al. Gastric activity and gut peptides in patients with functional dyspepsia: postprandial distress syndrome versus epigastric pain syndrome[J]. J Clin Gastroenterol, 2017,51(2):136-144.
[24]	Lunding JA, Tefera S, Gilja OH, et al. Rapid initial gastric emptying and hypersensitivity to gastric filling in functional dyspepsia: effects of duodenal lipids[J]. Scand J Gastroenterol,2006,41(9):1028-1036.
[25]	Kusano M, Zai H, Shimoyama Y, et al. Rapid gastric emptying, rather than delayed gastric emptying, might provoke functional dyspepsia[J]. J Gastroenterol Hepatol,2011,26(Suppl 3):75-78.
[26]	Koutouratsas T, Kalli T, Karamanolis G, et al. Contribution of ghrelin to functional gastrointestinal disorders’ pathogenesis[J]. World J Gastroenterol,2019,25(5):539-551.
[27]	Nwokolo CU, Freshwater DA, O'Hare P, et al. Plasma ghrelin following cure of Helicobacter pylori[J]. Gut,2003,52(5):637-640.
[28]	Akalu Y, Molla MD, Dessie G, et al. Physiological effect of ghrelin on body systems[J]. Int J Endocrinol,2020,2020:1385138.
[29]	Mantero P, Matus GS, Corti RE, et al. Helicobacter pylori and corpus gastric pathology are associated with lower serum ghrelin[J]. World J Gastroenterol,2018,24(3):397-407.

[1]	樊恒, 孙敏, 朱建华. 红景天苷通过抑制PI3K/AKT/mTOR信号通路对大鼠脓毒症急性肾损伤的保护作用[J/OL]. 中华危重症医学杂志(电子版), 2024, 17(03): 188-195.
[2]	崔玉峰, 林毅军, 王志民. 幽门螺杆菌分子检测技术研究进展[J/OL]. 中华实验和临床感染病杂志(电子版), 2024, 18(05): 257-262.
[3]	郭伟仪, 林沛玲. 不同抗体型幽门螺杆菌感染与溃疡性结肠炎患者疾病活动及组织学评分的关系[J/OL]. 中华实验和临床感染病杂志(电子版), 2024, 18(04): 237-244.
[4]	沈华娟, 庄剑波, 刘春. 幽门螺杆菌感染抗体分型与胃黏膜炎性病变程度及黏膜组织学变化间的相关性[J/OL]. 中华实验和临床感染病杂志(电子版), 2024, 18(03): 156-162.
[5]	杜成旭, 李冬瑞, 张树彬, 李秋生, 邢中强, 王天阳, 赵伟红, 刘建华. 新胰胃吻合在腹腔镜胰腺中段切除术中的应用价值[J/OL]. 中华肝脏外科手术学电子杂志, 2024, 13(04): 504-508.
[6]	伍宵, 潘立, 谢理政, 茆翔. 幽门螺杆菌感染和循环miRNA变化与老年脑干出血患者应激性溃疡的关系[J/OL]. 中华神经创伤外科电子杂志, 2024, 10(01): 42-47.
[7]	杨卫东, 周威, 向洪涛. 慢性萎缩性胃炎患者幽门螺杆菌感染与炎性细胞因子及病理特征的关系[J/OL]. 中华消化病与影像杂志(电子版), 2024, 14(05): 459-464.
[8]	李艳, 邹联琼, 刘千喜, 徐寅. 隔药饼脐灸对功能性消化不良患者中医症状积分、胃电图参数及胃排空率的影响[J/OL]. 中华消化病与影像杂志(电子版), 2024, 14(05): 474-477.
[9]	窦丽辉, 李鹏, 窦静敏, 王贵玲. 辨证针灸联合双歧杆菌三联活菌胶囊治疗脑梗死恢复期脾胃虚弱证功能性消化不良的疗效[J/OL]. 中华消化病与影像杂志(电子版), 2024, 14(03): 218-221.
[10]	汤峥丽, 王芳, 王唯坚. 中老年人群幽门螺杆菌感染对非酒精性脂肪肝及冠状动脉粥样硬化影响的关联性分析[J/OL]. 中华消化病与影像杂志(电子版), 2024, 14(02): 137-140.
[11]	何妍, 檀丽冰, 周雯, 魏敏, 连军松. 老年男性人群幽门螺杆菌感染与血脂谱的关联研究[J/OL]. 中华胃肠内镜电子杂志, 2024, 11(03): 171-175.
[12]	李志伟, 汤泊夫, 孙鹤, 贾语婧, 江振宇, 刘琳, 李慧, 党彤. 利用探头式共聚焦激光显微内镜荧光定量评价除菌治疗对幽门螺杆菌相关性萎缩性胃炎黏膜屏障功能影响的研究[J/OL]. 中华胃肠内镜电子杂志, 2024, 11(02): 94-99.
[13]	冯建聪, 普布扎西, 大旦增, 宋虎, 彭学成, 程志立, 林杨, 张昊翔. 西藏山南地区幽门螺杆菌感染的单中心分析[J/OL]. 中华胃肠内镜电子杂志, 2024, 11(01): 37-40.
[14]	薛黎, 高孝忠. 幽门螺杆菌阳性患者服药依从性预测模型的建立[J/OL]. 中华胃肠内镜电子杂志, 2024, 11(01): 23-27.
[15]	蒋琦, 刘卫东, 艾合买江·库尔班江, 惠文佳, 张梦霞, 李紫琼, 高峰. 不同类型幽门螺杆菌合并反流性食管炎的临床病理特征分析[J/OL]. 中华胃食管反流病电子杂志, 2024, 11(02): 95-99.

阅读次数

全文

摘要

选择文件类型/文献管理软件名称

选择包含的内容

Animal experimental study of Helicobacter pylori associated dyspepsia mediated by ghrelin

模态框（Modal）标题

选择文件类型/文献管理软件名称

选择包含的内容

Animal experimental study of Helicobacter pylori associated dyspepsia mediated by ghrelin