切换至 "中华医学电子期刊资源库"

中华卫生应急电子杂志 ›› 2022, Vol. 08 ›› Issue (04) : 216 -221. doi: 10.3877/cma.j.issn.2095-9133.2022.04.005

论著

大黄附子汤改善小鼠重症急性胰腺炎肠动力障碍的机制研究
宋轶1, 仲益龙2, 路晓光1,(), 战丽彬3, 范治伟1, 路童4, 陈海铭4, 芮庆林5   
  1. 1. 116001 辽宁大连,大连大学附属中山医院急诊医学科
    2. 116622 辽宁大连,大连大学研究生院
    3. 110847 辽宁沈阳,辽宁中医药大学中医药创新工程技术中心
    4. 110847 辽宁沈阳,辽宁中医药大学附属医院急诊医学科
    5. 210004 江苏南京,江苏省中医院急诊医学科
  • 收稿日期:2022-04-18 出版日期:2022-08-18
  • 通信作者: 路晓光
  • 基金资助:
    国家自然科学基金(81673801,81473512)

Mechanism of Dahuang Fuzi decoction in alleviating intestinal motility disorder in mice with severe acute pancreatitis

Yi Song1, Yilong Zhong2, Xiaoguang Lu1,(), Libin Zhan3, Zhiwei Fan1, Tong Lu4, Haiming Chen4, Qinglin Rui5   

  1. 1. Department of Emergency Medicine, the Affiliated Zhongshan Hospital of Dalian University, Dalian 116001, China
    2. Graduate School of Dalian University, Dalian 116622, China
    3. Innovation Engineering and Technology Center of Traditional Chinese Medicine, Liaoning University of Traditional Chinese Medicine, Shenyang 110847, China
    4. Department of Emergency Medicine, the Affiliated Hospital of Liaoning University of Traditional Chinese Medicine, Shenyang 110847, China
    5. Department of Emergency Medicine, Jiangsu Hospital of Traditional Chinese Medicine, Nanjing 210004, China
  • Received:2022-04-18 Published:2022-08-18
  • Corresponding author: Xiaoguang Lu
引用本文:

宋轶, 仲益龙, 路晓光, 战丽彬, 范治伟, 路童, 陈海铭, 芮庆林. 大黄附子汤改善小鼠重症急性胰腺炎肠动力障碍的机制研究[J/OL]. 中华卫生应急电子杂志, 2022, 08(04): 216-221.

Yi Song, Yilong Zhong, Xiaoguang Lu, Libin Zhan, Zhiwei Fan, Tong Lu, Haiming Chen, Qinglin Rui. Mechanism of Dahuang Fuzi decoction in alleviating intestinal motility disorder in mice with severe acute pancreatitis[J/OL]. Chinese Journal of Hygiene Rescue(Electronic Edition), 2022, 08(04): 216-221.

目的

揭示大黄附子汤改善重症急性胰腺炎小鼠早期肠动力障碍的机制。

方法

将27只SPF级雄性C57BL/6小鼠随机分为对照组、重症急性胰腺炎组(SAP)和大黄附子汤治疗组(DHFZT),每组9只。SAP组和DHZFT组采用腹腔注射20% L-精氨酸溶液复制SAP小鼠模型,对照组腹腔注射等渗盐水。DHFZT组分别于造模后12、24、36 h以0.2 mL大黄附子汤灌肠,对照组和SAP组分别以0.2 mL等渗盐水灌肠。造模后48 h,每组取3只小鼠测定小肠推进率,其余小鼠麻醉后取血、取材。ELISA法检测小鼠血清淀粉酶、血钙、内毒素、TNF-α和IL-6含量。HE染色观察胰腺和回肠组织病理学变化。Western blot定量检测回肠nNOS、ChAT、VIP、SP蛋白表达。

结果

与对照组相比,SAP组的抑制性神经元nNOS和VIP数量显著增加,兴奋性神经元ChAT和SP数量显著下降,小肠推进率显著降低,对应血清淀粉酶、内毒素、TNF-α和IL-6水平显著升高,差异有统计学意义(P均<0.05);与SAP组相比,DHFZT组的抑制性神经元nNOS和VIP数量显著下降,而兴奋性神经元ChAT和SP数量显著增加,小肠推进率显著增加,对应血清淀粉酶、内毒素、TNF-α和IL-6水平显著降低,血钙含量显著上升,差异有统计学意义(P均<0.05)。

结论

SAP小鼠早期发生肠动力障碍的机制可能存在肠神经系统的重塑,表现在抑制性神经元数量增加、兴奋性神经元数量减少,肠动力受抑制;大黄附子汤能够逆转肠神经系统的重塑,从而促进SAP小鼠肠动力的恢复。

Objective

To reveal the possible mechanism of intestinal motility disorder in mice with severe acute pancreatitis (SAP) and the effect of Dahuangfuzi decoction on it.

Methods

Twenty seven SPF male C57BL/6 mice were randomly divided into control group, severe acute pancreatitis group (SAP) and rhubarb and aconite decoction treatment group (DHFZT), with 9 mice in each group. SAP group and DHZFT group were intraperitoneally injected with 20% L-arginine solution to replicate SAP mouse model, and the control group was intraperitoneally injected with isotonic saline. In DHFZT group, 0.2 mL rhubarb and aconite decoction was used for enema 12, 24 and 36 hours after modeling, while 0.2 mL isotonic saline was used for enema in control group and SAP group. At 48h after modeling, 3 mice in each group were taken to measure the small intestine propulsion rate, and the rest of the mice were taken blood and materials after anesthesia. Detection of serum amylase, blood calcium, endotoxin and TNF in mice by ELISA- α And IL-6 content. Histopathological changes of pancreas and ileum were observed with HE staining. Western blot was used to detect the expression of nNOS, ChAT, VIP and SP proteins in ileum.

Results

Compared with the control group, the expression of nNOS and VIP in the SAP group were increased significantly, and the ChAT and SP were decreased significantly; the propulsion rate of small intestine was decreased significantly, with the lower levels of serum amylase, endotoxin, TNF-α and IL-6, and the blood calcium content also significantly decreased. Compared with the SAP group, the expression of nNOS and VIP in the DHFZT group were decreased significantly, and the ChAT and SP were increased significantly; the propulsion rate of small intestine was increased significantly, with the lower levels of serum amylase, endotoxin, TNF-α and IL-6, and the blood calcium content also significantly increased.

Conclusion

The mechanism of intestinal motility disorder in the early course of SAP mice may be related to the ENS remodeling, the increased number of inhibitory neurons, the decreased number of excitatory neurons, and therefore the proportion was imbalanced, leading to intestinal motility inhibition. Dahuangfuzi decoction may reverse the remodeling of ENS neurons and promote the recovery of intestinal motility in SAP mice.

表1 各组小鼠血清淀粉酶、血钙含量(±s)
表2 血清内毒素、TNF-α和IL-6水平含量(±s)
图1 小鼠胰腺、回肠HE染色(200×)注:SAP为重症胰腺炎组,DHFZT为大黄附子汤治疗组
图2 Western blot检测各组小鼠回肠组织nNOS、ChAT、VIP、SP蛋白表达注:nNOS为一氧化氮合酶,ChAT为胆碱乙酰转移酶,VIP为血管活性肠肽,SP为P物质;SAP为重症胰腺炎组,DHFZT为大黄附子汤治疗组;SAP组与对照组相比:aP<0.05,治疗组与SAP组相比:bP<0.05
1
中华医学会外科学分会胰腺外科学组.中国急性胰腺炎诊治指南(2021)[J].中华消化外科杂志202120(7):730-739.
2
夏玫,龚建平.重症急性胰腺炎合并腹腔间隔室综合征的外科治疗进展[J].国际外科学杂志201946(9):640-642.
3
王旭,邓颖.腹腔内高压的相关研究进展[J].医学综述201925(8):1625-1629.
4
廖辰,姚壮凯.腹腔间隔室综合征的诊疗进展[J].临床外科杂志201624(5):398-400.
5
温聪聪,王颖,吴德卿,等.重症急性胰腺炎胃肠功能障碍的诊治和发生机制研究进展[J].中华胰腺病杂志201919(2):143-145.
6
Bressan AK, Ball CG.Intra-abdominal hypertension and abdominal compartment syndrome in acute pancreatitis,hepato-pancreato-biliary operations and livertransplantation[J].Anaesthesiol Intensive Ther2017, 49(2):159-166.
7
路小光,战丽彬,曲明阳,等.大黄附子汤对重症急性胰腺炎大鼠细胞因子的影响[J].中国中西医结合急救杂志200411(6):352-354.
8
Kang X, Lu XG, Zhan LB,et al.Dai-Huang-Fu-Zi-Tang alleviates pulmonary and intestinal injury with severe acute pancreatitis via regulating aquaporins in rats[J].BMC Complement Altern Med201717(1):288.
9
宋轶,刘杰,路晓光,等.大黄附子汤改善重症急性胰腺炎大鼠肠道动力障碍的机制研究[J].中华急诊医学杂志202130(8):954-959.
10
吕春雨,路小光,战丽彬,等.大黄附子汤治疗重症急性胰腺炎合并急性呼吸窘迫综合征患者的疗效分析[J].中华急诊医学杂志201322(4):404-408.
11
Swanson JC, Chapin LT, Hankenson FC.Management of animal care and use programs in research, education, and testing[M].2th ed. Boca Raton (FL): CRC Press/Taylor &Francis,2018:23.
12
Gecelter G, Fahoum B, Gardezi S,et al.Abdominal compartment syndrome in severe acute pancreatitis:an indication for a decompressing laparotomy?[J].Dig Surg200219(5):402-405.
13
王鹏飞,李幼生.重症急性胰腺炎的营养治疗:肠衰竭与肠苏醒[J].腹部外科202134(5):336-339,346.
14
陈鹏德,王宝西.Cajal间质细胞相关研究进展[J].中华实用儿科临床杂志201631(19):1506-1508.
15
Nagy N, Goldstein AM.Enteric nervous system development:a crest cell’s journey from neural tube to colon[J].Semin Cell Dev Biol2017(66):94-106.
16
Sasselli V, Pachnis V, Burns AJ.The enteric nervous system[J].Dev Biol, 2012366(1):64-73.
17
Wood JD.Effects of bacteria on the enteric nervous system:implications for the irritable bowel syndrome[J].J Clin Gastroenterol2007(41 Suppl 1):S7-S19.
18
Kunze WA, Furness JB.The enteric nervous system and regulation of intestinal motility[J].Annu Rev Physiol1999(61):117-142.
19
Lin Z, Sandgren K, Ekblad E.Increased expression of vasoactive intestinal polypeptide in cultured myenteric neurons from adult rat small intestine[J].Auton Neurosci2003107(1):9-19.
20
Lin Z, Sandgren K, Ekblad E.Increased expression of nitric oxide synthase in cultured neurons from adult rat colonic submucous ganglia[J].Auton Neurosci2004114(1-2):29-38.
21
Ekelund KM, Ekblad E.Structural,neuronal,and functional adaptive changes in atrophic rat ileum[J].Gut199945(2):236-245.
22
McQuade RM, Carbone SE, Stojanovska V,et al.Role of oxidative stress in oxaliplatin-induced enteric neuropathy and colonic dysmotility in mice[J].Br J Pharmacol2016173(24):3502-3521.
23
Lin Z, Liu Y, Zheng Q,et al.Increased proportion of nitric oxide synthase immunoreactive neurons in rat ileal myenteric ganglia after severe acute pancreatitis[J].BMC Gastroenterol2011(11):127.
[1] 陆婷, 范晴敏, 王洁, 万晓静, 许春芳, 董凤林. 超声引导下经皮穿刺置管引流对重症急性胰腺炎的疗效及应用时机的选择[J/OL]. 中华医学超声杂志(电子版), 2024, 21(05): 511-516.
[2] 宫丹丹, 孙飞飞, 于健, 姜晓东. 重症急性胰腺炎死亡风险因素分析及风险评估模型建立[J/OL]. 中华危重症医学杂志(电子版), 2024, 17(01): 19-25.
[3] 孙亚慧, 李甜甜. 在重症急性胰腺炎患者继发感染中调节性T细胞/辅助性T细胞17失衡的机制研究[J/OL]. 中华危重症医学杂志(电子版), 2023, 16(06): 475-480.
[4] 杨晶, 高青. 重症急性胰腺炎继发脓毒症的危险因素分析[J/OL]. 中华危重症医学杂志(电子版), 2023, 16(02): 105-110.
[5] 王晓梅, 刘冰, 马丽琼, 卢祖静, 苗建军. 基于LASSO-Cox回归分析的非轻症急性胰腺炎死亡风险列线图预测模型的建立和临床应用效果分析[J/OL]. 中华普通外科学文献(电子版), 2024, 18(01): 44-50.
[6] 黄翠君, 张喜玲, 刘思嘉, 刘云建. 重症急性胰腺炎营养支持治疗研究进展[J/OL]. 中华普通外科学文献(电子版), 2023, 17(05): 385-390.
[7] 党军强, 杨雁灵, 汪庆强, 尚琳, 朱磊, 项红军. 主动经皮穿刺引流治疗重症急性胰腺炎并发急性坏死物积聚的疗效分析[J/OL]. 中华普外科手术学杂志(电子版), 2024, 18(06): 671-674.
[8] 陈慧, 范婷, 李春花, 黄其密, 陈晓英, 翁启明, 彭茹, 唐思, 何佳霖, 陈庆, 王旖旎, 汤玲一. 重症急性胰腺炎并发肺部感染临床分析[J/OL]. 中华肺部疾病杂志(电子版), 2023, 16(06): 856-859.
[9] 高军龙, 张昕, 周倩倩, 袁媛. 重症急性胰腺炎早期免疫抑制的研究进展[J/OL]. 中华重症医学电子杂志, 2023, 09(03): 286-291.
[10] 宋燕秋, 戚桂艳, 杨双双, 周萍. 重症急性胰腺炎肠道菌群特征及早期肠内营养联合微生态制剂治疗的临床价值[J/OL]. 中华消化病与影像杂志(电子版), 2024, 14(05): 442-447.
[11] 丛黎, 马林, 陈旭, 李文文, 张亮亮, 周华亭. 改良CT严重指数联合炎症指标在重症急性胰腺炎患者胰腺感染预测及预后评估中的研究[J/OL]. 中华消化病与影像杂志(电子版), 2024, 14(05): 432-436.
[12] 汪纾羽, 焦茹, 石运涛. 早期肠内营养和微生态免疫肠内营养对重症急性胰腺炎患者肾损伤的预防效果及影响因素[J/OL]. 中华消化病与影像杂志(电子版), 2024, 14(02): 132-136.
[13] 王小红, 钱晶, 翁文俊, 周国雄, 朱顺星, 祁小鸣, 刘春, 王萍, 沈伟, 程睿智, 秦璟灏. 巯基丙酮酸硫基转移酶调控核因子κB信号介导自噬对重症急性胰腺炎大鼠的影响及机制[J/OL]. 中华消化病与影像杂志(电子版), 2023, 13(06): 422-426.
[14] 马强, 李军, 苟丽娟. 重症急性胰腺炎miR-21-3p、RUNX3表达水平及对病情发展程度的预测[J/OL]. 中华消化病与影像杂志(电子版), 2023, 13(05): 337-341.
[15] 王迎芝, 张先进, 杨士彦. 益生菌结合早期肠内营养对重症急性胰腺炎患者营养状况、肠黏膜屏障功能及肠内营养耐受性的影响[J/OL]. 中华消化病与影像杂志(电子版), 2023, 13(01): 55-58.
阅读次数
全文


摘要