Effect of <italic style="font-style: italic">Yigan Fupi Prescription</italic> （抑肝扶脾方） on the AKT/mTOR Pathway in the Colon Tissue of Diarrhea-Type Irritable Bowel Syndrome Model Rats with Liver Stagnation and Spleen Deficiency Pattern

JIU Suting; LI Huiying; SUN Yueting; CUI Songxiang; XU Xintian; ZHENG Hao; LU Weimin

doi:10.13288/j.11-2166/r.2025.03.013

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Effect of Yigan Fupi Prescription （抑肝扶脾方） on the AKT/mTOR Pathway in the Colon Tissue of Diarrhea-Type Irritable Bowel Syndrome Model Rats with Liver Stagnation and Spleen Deficiency Pattern

更新时间：2025-01-26

- Effect of Yigan Fupi Prescription （抑肝扶脾方） on the AKT/mTOR Pathway in the Colon Tissue of Diarrhea-Type Irritable Bowel Syndrome Model Rats with Liver Stagnation and Spleen Deficiency Pattern
- Journal of Traditional Chinese Medicine Vol. 66, Issue 3, Pages: 290-299(2025)
- 作者机构：
  
  1.南京中医药大学附属医院，江苏省南京市秦淮区汉中路155号，210029
  2.云南省曲靖市中医院
  3.南京市浦口区中医院
- 作者简介：
- 基金信息：
- DOI：10.13288/j.11-2166/r.2025.03.013
  CLC：
- Received：30 September 2024，
  
  Revised：2024-12-02，
  
  Published：02 February 2025
- 稿件说明：
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酒苏婷,李慧颖,孙月婷等.抑肝扶脾方对肝郁脾虚型腹泻型肠易激综合征模型大鼠结肠组织AKT/mTOR通路的影响[J].中医杂志,2025,66(03):290-299.

JIU Suting,LI Huiying,SUN Yueting,et al.Effect of Yigan Fupi Prescription （抑肝扶脾方） on the AKT/mTOR Pathway in the Colon Tissue of Diarrhea-Type Irritable Bowel Syndrome Model Rats with Liver Stagnation and Spleen Deficiency Pattern[J].Journal of Traditional Chinese Medicine,2025,66(03):290-299.
酒苏婷,李慧颖,孙月婷等.抑肝扶脾方对肝郁脾虚型腹泻型肠易激综合征模型大鼠结肠组织AKT/mTOR通路的影响[J].中医杂志,2025,66(03):290-299. DOI： 10.13288/j.11-2166/r.2025.03.013.

JIU Suting,LI Huiying,SUN Yueting,et al.Effect of Yigan Fupi Prescription （抑肝扶脾方） on the AKT/mTOR Pathway in the Colon Tissue of Diarrhea-Type Irritable Bowel Syndrome Model Rats with Liver Stagnation and Spleen Deficiency Pattern[J].Journal of Traditional Chinese Medicine,2025,66(03):290-299. DOI： 10.13288/j.11-2166/r.2025.03.013.

摘要

目的

从蛋白激酶B（AKT）/雷帕霉素靶蛋白（mTOR）通路探究抑肝扶脾方治疗肝郁脾虚型腹泻型肠易激综合征（IBS-D）的可能作用机制。

方法

将60只SD大鼠随机分为对照组，模型组，抑肝扶脾方低、中、高剂量组，匹维溴铵组，每组10只。除对照组外其余各组通过夹尾刺激21天+番泻叶灌胃14天的方法建立肝郁脾虚型IBS-D大鼠模型。造模成功后抑肝扶脾方低、中、高剂量组给药量分别为0.96、1.93、3.87 g/（kg·d），匹维溴铵组给药量为13.5 mg/（kg·d），对照组及模型组予10 ml/（kg·d）蒸馏水，各组均每天灌胃1次，连续14天。实验期间记录各组大鼠一般情况，造模后及给药后比较大鼠体质量、Bristol粪便评分、腹壁撤退反射实验（AWR）评分；HE染色观察大鼠结肠组织病理形态；ELISA法检测大鼠血清中肿瘤坏死因子α （TNF-α）、白细胞介素1β（IL-1β）、白细胞介素6（IL-6）水平；免疫荧光法检测大鼠结肠组织中AKT/mTOR通路相关蛋白［包括磷酸化蛋白激酶B（p-AKT）/AKT、磷酸化雷帕霉素靶蛋白（p-mTOR）/mTOR］相对表达量；Western Blot法检测大鼠结肠组织中自噬相关蛋白［包括UNC-51样激酶（ULK1）、Beclin1（自噬关键分子酵母Atg6同系物1）、微管相关蛋白1轻链3（LC3）］以及紧密连接蛋白［包括闭合蛋白（Occludin）、闭锁小带蛋白1（ZO-1）］水平。

结果

造模后，与对照组比较，其余各组大鼠体质量降低，粪便Bristol评分以及20、40、60、80 mmHg压力级别下AWR评分均升高（

＜0.05或

＜0.01）。给药后，与对照组比较，模型组大鼠体质量以及结肠组织中ULK1、Beclin1、LC3Ⅱ/LC3Ⅰ、Occludin、ZO-1蛋白水平均降低（

＜0.05或

＜0.01）；粪便Bristol评分，20、40、60、80 mmHg压力级别下AWR评分，血清中TNF-α、IL-1β、IL-6水平，以及结肠组织中p-AKT/AKT、p-mTOR/mTOR蛋白相对表达量均升高（

＜0.05或

＜0.01）；病理形态结果显示，结肠组织固有层腺体上部杯状细胞明显减少，少量炎性细胞浸润；黏膜下层胶原纤维溶解，分界不清，着色变淡，微血管充血、扩张。与模型组比较，抑肝扶脾方各剂量组及匹维溴铵组大鼠体质量，以及结肠组织中Beclin1、Occludin、LC3Ⅱ/LC3Ⅰ蛋白水平均升高（

＜0.05或

＜0.01）；粪便Bristol评分，40、60、80 mmHg压力级别下AWR评分，血清中IL-1β、IL-6、TNF-α水平，以及结肠组织中p-AKT/AKT、p-mTOR/mTOR蛋白相对表达量均降低（

＜0.05或

＜0.01）；抑肝扶脾方各剂量组及匹维溴铵组病理形态有不同程度的改善。与匹维溴铵组比较，抑肝扶脾方低、中剂量组在20、40、60 mmHg压力级别下AWR评分均升高，抑肝扶脾方低剂量血清中TNF-α、IL-1β、IL-6水平降低，抑肝扶脾方各剂量组结肠组织中p-mTOR/mTOR蛋白相对表达量均升高（

＜0.05）。与抑肝扶脾方低剂量组比较，抑肝扶脾方高剂量组及匹维溴铵组在各压力级别下AWR评分、结肠组织中p-AKT/AKT蛋白相对表达量降低，抑肝扶脾方中、高剂量组血清中TNF-α、IL-

1β水平升高，结肠组织中p-mTOR/mTOR蛋白相对表达量降低（

＜0.05）。

结论

抑肝扶脾方可有效改善肝郁脾虚型IBS-D模型大鼠结肠组织病理损伤，降低粪便Bristol及AWR评分，其作用机制可能与降低炎症因子水平，抑制结肠组织中AKT/mTOR通路相关蛋白，从而提高自噬相关蛋白水平有关。

Abstract

Objective

To explore the possible mechanism of

Yigan Fupi Prescription

（抑肝扶脾方， YFP） in treating diarrhea-type irritable bowel syndrome （IBS-D） by investigating the AKT/mTOR signaling pathway.

Methods

Sixty SD rats were randomly divided into control group， model group， YFP low-， medium-， and high-dose group， and pinaverium bromide group， with 10 rats in each group. All groups but the control group， were subjected to 21 days of tail-clamping stimulation and 14 days of senna leaf gavage to establish a liver stagnation and spleen deficiency-type IBS-D rat model. After successful modeling， the YFP low-， medium-， and high-dose group were administered 0.96， 1.93， and 3.87 g/（kg·d） of the prescription， respectively. The pinaverium bromide group was given 13.5 mg/（kg·d）， while the control and model groups were given 10 ml/（kg·d） distilled water. All groups were administered once daily for 14 consecutive days. General conditions of the rats were recorded during the experiment， and after modeling and drug administration， body weight， Bristol stool score， abdominal withdrawal reflex （AWR） score， and histo pathology of colon tissue were observed under HE staining. ELISA was used to detect serum levels of tumor necrosis factor α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-6 （IL-6）. Immunofluorescence was employed to detect the levels of AKT/mTOR pathway-related proteins including phosphorylated AKT （p-AKT）/AKT and phosphorylated mTOR （p-mTOR）/mTOR in the colon tissue. Western Blotting was used to detect the levels of autophagy-related proteins， including UNC-51-like kinase 1 （ULK1）， Beclin1 and LC3， and tight junction proteins including Occludin and ZO-1 in the colon tissue.

Results

After modeling， compared to the control group， the body weight of rats in the other groups decreased， and Bristol stool scores， as well as AWR scores under 20， 40， 60， and 80 mmHg increased （

＜0.05 or

＜0.01）. After drug administration， compared to the control group， the model group showed reduced body weight， decreased ULK1， Beclin1， LC3Ⅱ/LC3Ⅰ， Occludin， and ZO-1 protein levels in the colon tissue （

＜0.05 or

＜0.01）， and increased Bristol stool scores， AWR scores， serum TNF-α， IL-1β， and IL-6 levels， as well as p-AKT/AKT and p-mTOR/mTOR protein relative expression levels （

＜0.05 or

＜0.01）. Pathological results showed a significant reduction in goblet cells in the upper part of the glandular layer of the colon， with mild inflammatory cell infiltration. The submucosal collagen fibers were dissolved， with unclear boundaries， pale staining， and microvascular congestion and dilation. Compared with the model group， the YFP low-， medium-， and high-dose group and the pinaverium bromide group showed increased body weight， Beclin1， Occludin， and LC3Ⅱ/LC3Ⅰ protein levels （

＜0.05 or

＜0.01）， and decreased Bristol stool scores， AWR scores under 40， 60， and 80 mmHg， serum IL-1β， IL-6， TNF-α levels， and p-AKT/AKT， p-mTOR/mTOR protein relative expression levels （

＜0.05 or

＜0.01）. The pathological morphology of the rats in the YFP groups and pinaverium bromide group showed varying degrees of improvement. Compared with the pinaverium bromide group， the YFP low- and medium-dose group showed increased AWR scores under 20， 40， and 60 mmHg （

＜0.05）. The YFP low-dose group had reduced TNF-α， IL-1β， and IL-6 levels， and increased p-mTOR/mTOR protein relative expression levels occured in all YFP groups （

＜0.05）. Compared with the YFP low-dose group， the YFP high-dose group and pinaverium bromide group

showed decreased AWR scores under different pressure levels and reduced p-AKT/AKT protein relative expression levels， while the YFP medium- and high-dose group had elevated serum TNF-α， IL-1β levels and reduced p-mTOR/mTOR protein relative expression levels （

＜0.05）.

Conclusion

YFP can effectively improve the pathological injury of colon tissue in IBS-D model rats with liver stagnation and spleen deficiency， reduce Bristol stool and AWR scores， and its mechanism may be related to reducing level of inflammatory factors and inhibiting AKT/mTOR pathway-related proteins in colon tissue， thereby enhancing the expression of autophagy-related proteins in the colon tissue.

关键词

Keywords

references

LACY BE ， MEARIN F ， CHANG L ， et al . Bowel disorders ［J］. Gastroenterology ， 2016 ， 150 （ 6 ）： 1393 - 1407 .

OKA P ， PARR H ， BARBERIO B ， et al . Global prevalence of irritable bowel syndrome according to Rome Ⅲ or Ⅳcriteria： a systematic review and meta-analysis ［J］. Lancet Gastroenterol Hepatol ， 2020 ， 5 （ 10 ）： 908 - 917 .

FORD AC ， SPERBER AD ， CORSETTI M ， et al . Irritable bowel syndrome ［J］. Lancet ， 2020 ， 396 （ 10263 ）： 1675 - 1688 .

JOVANI M ， FIORINO G ， DANESE S . Commentary：associations between immune activation， intestinal permeability and irritable bowel syndrome ［J］. Aliment Pharmacol Ther ， 2013 ， 37 （ 2 ）： 277 - 278 .

HANNING N ， EDWINSON AL ， CEULEERS H ， et al . Intestinal barrier dysfunction in irritable bowel syndrome：a systematic review ［J］. Therap Adv Gastroenterol ， 2021 ， 14 ： 1 - 31 . doi： 10.1177/1756284821993586 http://dx.doi.org/10.1177/1756284821993586 .

FENG Y ， TEITELBAUM DH . Tumour necrosis factor-induced loss of intestinal barrier function requires TNFR1 and TNFR2 signalling in a mouse model of total parenteral nutrition ［J］. J Physiol ， 2013 ， 591 （ 15 ）： 3709 - 3723 .

AL-SADI R ， YE D ， SAID HM ， et al . IL-1beta-induced increase in intestinal epithelial tight junction permeability is mediated by MEKK-1 activation of canonical NF-κB pathway ［J］. Am J Pathol ， 2010 ， 177 （ 5 ）： 2310 - 2322 .

ZHU ML ， ZHANG PM ， JIANG M ， et al . Myricetin induces apoptosis and autophagy by inhibiting PI3K/Akt/mTOR signalling in human colon cancer cells ［J］. BMC Complement Med Ther ， 2020 ， 20 （ 1 ）： 209 .

赵旦娅，郑浩，宋昌娟，等 . 抑肝扶脾方治疗肝郁脾虚证腹泻型肠易激综合征的临床研究［J］. 中华中医药杂志， 2022 ， 37 （ 12 ）： 7491 - 7494 .

赵双梅，李慧臻，孔祥茹，等 . 肝郁脾虚型IBS-D大鼠模型的建立及评价［J］. 湖南中医杂志， 2015 ， 31 （ 10 ）： 137 - 139 .

赵妍，罗丹妮，陈颖，等 . 慢性束缚应激联合番泻叶灌胃法制备IBS-D大鼠模型的量效及时效关系评价［J］. 世界华人消化杂志， 2017 ， 25 （ 15 ）： 1360 - 1367 .

LEWIS SJ ， HEATON KW . Stool form scale as a useful guide to intestinal transit time ［J］. Scand J Gastroenterol ， 1997 ， 32 （ 9 ）： 920 - 924 .

AL-CHAER ED ， KAWASAKI M ， PASRICHA PJ . A new model of chronic visceral hypersensitivity in adult rats induced by colon irritation during postnatal development ［J］. Gastroenterology ， 2000 ， 119 （ 5 ）： 1276 - 1285 .

徐叔云，卞如濂，陈修 . 药理实验方法学［M］. 3版 . 北京：人民卫生出版社， 2002 ： 1861 .

中华中医药学会脾胃病分会 . 肠易激综合征中医诊疗专家共识意见（2017）［J］. 中医杂志， 2017 ， 58 （ 18 ）： 1614 – 1620 .

徐景藩 . 徐景藩脾胃病临证经验集粹［M］. 陆为民，徐丹华，罗斐和，整理.北京：科学技术出版社， 2015 ： 40 - 42 .

曹正龙，何镔，潘军，等 . 国医大师徐景藩抑肝扶脾法治疗腹泻型肠易激综合征经验应用的研究［J］. 中医药临床杂志， 2013 ， 25 （ 7 ）： 604 - 606 .

MARTÍNEZ C ， LOBO B ， PIGRAU M ， et al . Diarrhoea-predominant irritable bowel syndrome： an organic disorder with structural abnormalities in the jejunal epithelial barrier ［J］. Gut ， 2013 ， 62 （ 8 ）： 1160 - 1168 .

SAHA K ， SUBRAMENIUM GANAPATHY A ， WANG A ， et al . Autophagy reduces the degradation and promotes membrane localization of Occludin to enhance the intestinal epithelial tight junction barrier against paracellular macromolecule flux ［J］. J Crohns Colitis ， 2023 ， 17 （ 3 ）： 433 - 449 .

WONG M ， GANAPATHY AS ， SUCHANEC E ， et al . Intestinal epithelial tight junction barrier regulation by autophagy-related protein ATG6/beclin 1 ［J］. Am J Physiol Cell Physiol ， 2019 ， 316 （ 5 ）： C753 - C765 .

PETERSON LW ， ARTIS D . Intestinal epithelial cells： regulators of barrier function and immune homeostasis ［J］. Nat Rev Immunol ， 2014 ， 14 （ 3 ）： 141 - 153 .

OHMAN L ， SIMRÉN M . Pathogenesis of IBS： role of inflammation， immunity and neuroimmune interactions ［J］. Nat Rev Gastroenterol Hepatol ， 2010 ， 7 （ 3 ）： 163 - 173 .

POLIVKA J JR ， JANKU F . Molecular targets for cancer therapy in the PI3K/AKT/mTOR pathway ［J］. Pharmacol Ther ， 2014 ， 142 （ 2 ）： 164 - 175 .

YU X ， LONG YC ， SHEN HM . Differential regulatory functions of three classes of phosphatidylinositol and phosphoinositide 3-kinases in autophagy ［J］. Autophagy ， 2015 ， 11 （ 10 ）： 1711 - 1728 .

CHEN Y ， KLIONSKY DJ . The regulation of autophagy- unanswered questions ［J］. J Cell Sci ， 2011 ， 124 （ Pt 2 ）： 161 - 170 .

KLIONSKY DJ ， ABDELMOHSEN K ， ABE A ， et al . Guidelines for the use and interpretation of assays for monitoring autophagy （3rd edition）［J］. Autophagy ， 2016 ， 12 （ 1 ）： 1 - 222 .

MIZUSHIMA N ， KOMATSU M . Autophagy： renovation of cells and tissues ［J］. Cell ， 2011 ， 147 （ 4 ）： 728 - 741 .

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Effect of Yigan Fupi Prescription （抑肝扶脾方） on the AKT/mTOR Pathway in the Colon Tissue of Diarrhea-Type Irritable Bowel Syndrome Model Rats with Liver Stagnation and Spleen Deficiency Pattern

DOI：10.13288/j.11-2166/r.2025.03.013

摘要

Abstract

关键词

Keywords

references