Guiqi Baizhu Prescription Combined with Oxaliplatin Protects Intestinal Barrier of Tumor-bearing Mice with Gastric Cancer by Regulation of VIP/cAMP/PKA/AQP Signaling Pathway

DONG Huancheng; SU Yun; GONG Hongxia; CAO Wangjie; HE Jianzheng; LIU Yongqi; ZHANG Han; ZENG Yuanding; LI Congyi; KANG Qian

doi:10.13422/j.cnki.syfjx.20222103

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Guiqi Baizhu Prescription Combined with Oxaliplatin Protects Intestinal Barrier of Tumor-bearing Mice with Gastric Cancer by Regulation of VIP/cAMP/PKA/AQP Signaling Pathway

更新时间：2023-02-07

- Guiqi Baizhu Prescription Combined with Oxaliplatin Protects Intestinal Barrier of Tumor-bearing Mice with Gastric Cancer by Regulation of VIP/cAMP/PKA/AQP Signaling Pathway
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 29, Issue 5, Pages: 129-136(2023)
- 作者机构：
  
  1.甘肃中医药大学基础医学院，兰州 730000
  2.甘肃省高校重大疾病分子医学与中医药防治研究省级重点实验室，兰州 730000
  3.甘肃省中医药防治慢性疾病重点实验室，兰州 730000
  4.国家卫生健康委胃肠肿瘤诊治重点实验室，甘肃省人民医院，兰州 730000
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20222103
  CLC： R2-0;R22;R285.5;R289;R33
- Received：25 August 2022，
  
  Published Online：19 November 2022，
  
  Published：05 March 2023
- 稿件说明：
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董焕成,苏韫,龚红霞等.归芪白术方联合奥沙利铂通过调节VIP/cAMP/PKA/AQPs信号通路保护胃癌荷瘤小鼠肠道屏障[J].中国实验方剂学杂志,2023,29(05):129-136.

DONG Huancheng,SU Yun,GONG Hongxia,et al.Guiqi Baizhu Prescription Combined with Oxaliplatin Protects Intestinal Barrier of Tumor-bearing Mice with Gastric Cancer by Regulation of VIP/cAMP/PKA/AQP Signaling Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(05):129-136.
董焕成,苏韫,龚红霞等.归芪白术方联合奥沙利铂通过调节VIP/cAMP/PKA/AQPs信号通路保护胃癌荷瘤小鼠肠道屏障[J].中国实验方剂学杂志,2023,29(05):129-136. DOI： 10.13422/j.cnki.syfjx.20222103.

DONG Huancheng,SU Yun,GONG Hongxia,et al.Guiqi Baizhu Prescription Combined with Oxaliplatin Protects Intestinal Barrier of Tumor-bearing Mice with Gastric Cancer by Regulation of VIP/cAMP/PKA/AQP Signaling Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(05):129-136. DOI： 10.13422/j.cnki.syfjx.20222103.

摘要

目的

探讨归芪白术方联合奥沙利铂通过血管活性肠肽（VIP）/环磷酸腺苷（cAMP）/蛋白激酶A（PKA）信号通路调控下游水通道蛋白3（AQP3）和水通道蛋白4（AQP4）从而保护胃癌荷瘤小鼠肠道屏障的作用。

方法

将密度为1×10

个/mL的胃癌细胞株MFC制成细胞悬液，经荷瘤小鼠右腋下接种细胞悬液0.2 mL，构建胃癌荷瘤小鼠模型，造模成功后将小鼠随机分为5组，模型组、奥沙利铂组（10 mg·kg

-1

）、奥沙利铂加归芪白术方高、中、低剂量组（17.68、8.84、4.42 g·kg

-1

），每组10只，另外余10只作为空白组。各组小鼠经灌胃或腹腔注射中药、奥沙利铂或生理盐水，治疗14 d。末次给药后，次日眼球取血分离血清并取其结肠样本。苏木素-伊红（HE）染色观察组织形态的变化。酶联免疫吸附测定法（ELISA）检测血清中

-乳酸（

-LA）、二胺氧化酶（DAO）含量；实时荧光定量聚合酶链式反应（Real-time PCR）及蛋白免疫印迹法（Western blot）分别检测各组小鼠结肠组织中VIP、cAMP、PKA、AQP3和AQP4 mRNA及蛋白的表达。

结果

与正常组比较，模型组黏膜下层水肿，黏膜层中肠腺排列紊乱，杯状细胞缺失，可见大量炎性细胞浸润及绒毛脱落的现象。而各给药组均有不同程度的改善；与正常组比较，模型组血清中DAO和

-LA水平均显著上升（

0.01），与模型组比较，联合用药高剂量组DAO和

-LA水平及联合用药中剂量组

-LA水平均有所下降（

0.05，

0.01），与奥沙利铂组比较，联合用药高、中剂量组

-LA水平有所下降（

0.05），其余组DAO和

-LA表达水平也有所下降，但差异无统计学意义；与正常组比较，模型组小鼠的VIP、cAMP、PKA、AQP3和AQP4 mRNA及蛋白表达水平明显降低（

0.05，

0.01）；与模型组比较，各给药组小鼠的VIP、cAMP、PKA、AQP3和AQP4 mRNA及蛋白表达水平均有所升高，其中联合用药高剂量组VIP、cAMP、PKA、AQP3和AQP4

mRNA及蛋白表达水平明显升高（

0.05，

0.01），联合用药中剂量组cAMP蛋白表达水平升高（

0.05）；与奥沙利铂组比较，联合用药高剂量组cAMP蛋白表达水平明显升高（

0.05），其余组mRNA及蛋白表达也有所升高，但差异无统计学意义。

结论

归芪白术方联合奥沙利铂通过VIP/cAMP/PKA信号通路调节AQP3和AQP4达到保护胃癌荷瘤小鼠肠道屏障的作用。

Abstract

Objective

To investigate the protective effect of Guiqi Baizhu prescription combined with oxaliplatin on the intestinal barrier of tumor-bearing mice with gastric cancer by regulating downstream aquaporin 3 （AQP3） and aquaporin 4 （AQP4） through the vasoactive intestinal peptide （VIP）/cyclic adenosine monophosphate （cAMP）/protein kinase A （PKA） signaling pathway.

Method

The gastric cancer cell lines MFC with a density of 1×10

/mL were prepared into cell suspension. The tumor-bearing mouse model of gastric cancer was established by inoculating 0.2 mL cell suspension under the right axilla of mice. After successful modeling， mice were randomly divided into 5 groups， namely， model group， oxaliplatin group （10 mg·kg

-1

）， and high， medium， and low-dose oxaliplatin + Guiqi Baizhu prescription groups （17.68， 8.84， 4.42 g·kg

-1

）， with 10 mice in each group， and the remaining 10 mice were set as a blank group. Mice in each group were treated with Chinese medicine， oxaliplatin， or normal saline by gavage or intraperitoneal injection for 14 d. The next day after the last dose， blood was taken from the eyeball to separate serum and take colonic samples. Hematoxylin-eosin （HE） staining was used to observe the changes in tissue morphology. The content of

-lactate acid （

-LA） and diamine oxidase （DAO） in the serum was determined by enzyme-linked immunosorbent assay （ELISA）. The mRNA and protein expressions of VIP， cAMP， PKA， AQP3， and AQP4 were detected by Real-time quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively.

Result

Compared with the blank group， the model group showed edema in the colonic submucosa， disordered arrangement of intestinal glands in the mucosal layer， loss of goblet cells， infiltration of inflammatory cells， and villus shedding. However， there were different degrees of improvement in each administration group. As compared with the blank group， the serum levels of DAO and

-LA in the model group were significantly increased （

0.01）. As compared with the model group， the levels of DAO and

-LA in the high-dose oxaliplatin + Guiqi Baizhu prescription group and the level of

-LA in the medium-dose oxaliplatin + Guiqi Baizhu prescription group were decreased （

0.05，

0.01）. As compared with the oxaliplatin group， the levels of

-LA in the high and medium-dose oxaliplatin + Guiqi Baizhu prescription groups were decreased （

0.05）， and the levels of DAO and

-LA in other administration groups were decreased as well， but the difference had no statistical significance. As compared with the blank group， the mRNA and protein expression levels of VIP， cAMP， PKA， AQP3， and AQP4 in the model group were significantly decreased （

0.05，

0.01）. As compared with the model group， the mRNA and protein expression levels of VIP， cAMP， PKA， AQP3， and AQP4 in each administration group were increased， and those in the high-dose oxaliplatin + Guiqi Baizhu prescription group were significantly increased （

0.05，

0.01）， while the protein expression level of cAMP in the medium-dose oxaliplatin + Guiqi Baizhu prescription group were increased （

0.05）. As compared with the oxaliplatin group， the protein expression levels of cAMP in the high-dose oxaliplatin + Guiqi Baizhu prescription group were increased （

0.05）， and the mRNA and protein expressions of these indexes in the other groups were also increased but the differences were not statistically significant.

Conclusion

Guiqi Baizhu prescription combined with oxaliplatin can regulate AQP3 and AQP4 through the VIP/cAMP/PKA signaling pathway to protect the intestinal barrier of tumor-bearing mice with gastric cancer.

关键词

Keywords

references

SUNG H ， FERLAY J ， SIEGEL R L ， et al . Global cancer statistics 2020：GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries ［J］. CA Cancer J Clin ， 2021 ， 71 （ 3 ）： 209 - 249 .

李翔子，李市荣，杨田野，等 . 首荟通便胶囊对慢性传输性便秘小鼠肠道屏障的影响［J］. 中草药， 2022 ， 53 （ 5 ）： 1458 - 1462 .

王宏，徐娟，江茜，等 . 大黄-丹参药对对慢性肾衰竭模型大鼠肠源性尿毒素含量和肠道屏障功能的影响［J］. 中国药房， 2021 ， 32 （ 7 ）： 825 - 831 .

牛世伟，苏韫，龚红霞，等 . 归芪白术方联合奥沙利铂对胃癌荷瘤小鼠机体免疫炎症分子的影响及抑癌作用［J］. 中国免疫学杂志， 2022 ， 38 （ 3 ）： 328 - 333 .

张晗，苏韫，龚红霞，等 . 归芪白术方联合奥沙利铂对胃癌荷瘤小鼠EGFR，VEGFR2表达和血管生成的影响［J］. 中国实验方剂学杂志， 2022 ， 28 （ 7 ）： 57 - 63 .

龚红霞，李婷婷，苏韫，等 . 归芪白术方含药血清通过PI3K/Akt信号通路对共培养的胃癌MKN-45细胞凋亡的影响［J］. 时珍国医国药， 2021 ， 32 （ 12 ）： 2890 - 2893 .

牛世伟，苏韫，龚红霞，等 . 胃癌患者血清IL-1的升高和癌组织JAK2/STAT3信号通路的激活［J］. 基础医学与临床， 2022 ， 42 （ 3 ）： 401 - 405 .

刘艳菊，刘景超，王永飞，等 . 绞股蓝多糖对MFC胃癌荷瘤小鼠肿瘤生长抑制及免疫调节作用［J］. 中成药， 2019 ， 41 （ 12 ）： 2876 - 2881 .

牛世伟 . 归芪白术方联合奥沙利铂调控IL-6/JAK2/STAT3信号通路对荷瘤小鼠胃癌细胞增殖及炎症免疫的影响［D］. 兰州：甘肃中医药大学， 2022 .

李春波，苏韫，薛轩，等 . 基于中药分子机制的生物信息学在线分析归芪白术方辅助治疗胃癌的作用机制［J］. 甘肃中医药大学学报， 2019 ， 36 （ 6 ）： 7 - 13 .

SUN X ， CUI Y ， SU Y ， et al . Dietary fiber ameliorates lipopolysaccharide-induced intestinal barrier function damage in piglets by modulation of intestinal microbiome ［J］. Msystems ， 2021 ， 6 （ 2 ）： e01374 - e01420 .

LI M ， RAO X ， CUI Y ， et al . The keratin 17/YAP/IL6 axis contributes to E-cadherin loss and aggressiveness of diffuse gastric cancer ［J］. Oncogene ， 2022 ， 41 （ 6 ）： 770 - 781 .

FILALY H E ， OUTLIOUA A ， MEDYOUF H ， et al . Targeting IL-1 β in patients with advanced Helicobacter pylori infection： A potential therapy for gastric cancer ［J］. Future Microbiol ， 2022 ， 17 （ 8 ）： 633 - 641 .

BASILE D ， DI NARDO P ， CORVAJA C ， et al . Mucosal injury during anti-cancer treatment：From pathobiology to bedside ［J］. Cancers ， 2019 ， 11 （ 6 ）： 857 .

PEARCE L R ， KOMANDER D ， ALESSI D R ， et al . The nuts and bolts of AGC protein kinases ［J］. Nat Rev Mol Cell Biol ， 2010 ， 11 （ 1 ）： 9 - 22 .

GANCEDO J M . Biological roles of cAMP：Variations on a theme in the different kingdoms of life ［J］. Biol Rev Camb Philos Soc ， 2013 ， 88 （ 3 ）： 645 - 668 .

WEI C X ， WU J H ， HUANG Y H ， et al . Lactobacillus plantarum improves LPS-induced Caco2 cell line intestinal barrier damage via cyclic AMP-PKA signaling ［J］. PLoS One ， 2022 ， 17 （ 5 ）： e0267831 .

ZHI X ， TAO J ， LI Z ， et al . MiR-874 promotes intestinal barrier dysfunction through targeting AQP3 following intestinal ischemic injury ［J］. FEBS letters ， 2014 ， 588 （ 5 ）： 757 - 763 .

HU R ， ZHANG T ， TANG F ， et al . Effect of Weichang'an pill on intestinal digestion enzymes and the AQP4 concentration in proximal colon in IBS-D rats ［J］. Chin J Chin Mat Med ， 2010 ， 35 （ 21 ）： 2899 - 2903 .

HUA Y ， DING S ， ZHANG W ， et al . Expression of AQP3 protein in hAECs is regulated by CAMP-PKA-CREB signalling pathway ［J］. Front Biosci ， 2015 ， 20 （ 7 ）： 1047 - 1055 .

WANG W B ， LI J T ， HUI Y ， et al . Combination of pseudoephedrine and emodin ameliorates LPS-induced acute lung injury by regulating macrophage M1/M2 polarization through the VIP/cAMP/PKA pathway ［J］. Chin Med ， 2022 ， 17 （ 1 ）： 1 - 17 .

MARMAROU C R ， LIANG X ， ABIDI N H ， et al . Selective vasopressin-1a receptor antagonist prevents brain edema，reduces astrocytic cell swelling and GFAP，V1aR and AQP4 expression after focal traumatic brain injury ［J］. Brain research ， 2014 ， 1581 ： 89 - 102 .

KITCHEN P ， DAY R E ， TAYLOR L H J ， et al . Identification and molecular mechanisms of the rapid tonicity-induced relocalization of the aquaporin 4 channel ［J］. J Biol Chem ， 2015 ， 290 （ 27 ）： 16873 - 16881 .

TAN Q ， HU J ， ZHOU Y ， et al . Inhibitory effect of Lactococcus lactis subsp.lactis HFY14 on diphenoxylate-induced constipation in mice by regulating the VIP-cAMP-PKA-AQP3 signaling pathway ［J］. Drug Des Devel Ther ， 2021 ， 15 ： 1971 .

李婷婷，苏韫，龚红霞，等 . 归芪白术汤对脾虚小鼠免疫因子及水通道蛋白表达的影响［J］. 时珍国医国药， 2021 ， 32 （ 4 ）： 805 - 808 .

GAN Y ， AI G ， WU J ， et al . Patchouli oil ameliorates 5-fluorouracil-induced intestinal mucositis in rats via protecting intestinal barrier and regulating water transport ［J］. J Ethnopharmacol ， 2020 ， 250 ： 112519 .

SUN S ， LIU W ， LI Y ， et al . CADM1 enhances intestinal barrier function in a rat model of mild inflammatory bowel disease by inhibiting the STAT3 signaling pathway ［J］. J Bioenerg Biomembr ， 2020 ， 52 （ 5 ）： 343 - 354 .

LI Q ， XU T ， ZHONG H ， et al . Impaired intestinal barrier in patients with obstructive sleep apnea ［J］. Sleep Breath ， 2021 ， 25 （ 2 ）： 749 - 756 .

ZHANG X ， PENG B ， ZHANG Y ， et al . Biliary drainage reduces intestinal barrier damage in obstructive jaundice by regulating autophagy ［J］. Contrast Media Mol Imaging ， 2022 ， doi： 10.1155/2022/3301330 http://dx.doi.org/10.1155/2022/3301330 .

周永学，王郁金，张红，等 . 血管活性肠肽对便秘大鼠排便及结肠组织中 VIP-cAMP-PKA-AQP3 信号通路的影响［J］. 中南大学学报：医学版， 2016 ， 41 （ 11 ）： 1175 - 1180 .

王璐，隋楠 . 基于大肠主津理论助阳通便膏对便秘模型小鼠结肠组织VIP-CAMP-PKA-AQP3通路的影响［J］. 中华中医药学刊， 2022 ， 40 （ 5 ）：147-151， 277 - 278 .

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