Aconiti Lateralis Radix Praeparata-Cinnamomi Cortex Combination Alleviates Slow Transit Constipation due to Yang Deficiency in Rats via Regulating VIP/cAMP/PKA/AQP Pathway in Colon

ZHAO Luona; YANG Yuanfeng; LI Yuchuan; ZHANG Yuanzhe

doi:10.13422/j.cnki.syfjx.20231212

您当前的位置：

首页 >

文章列表页 >

Aconiti Lateralis Radix Praeparata-Cinnamomi Cortex Combination Alleviates Slow Transit Constipation due to Yang Deficiency in Rats via Regulating VIP/cAMP/PKA/AQP Pathway in Colon

更新时间：2023-10-13

- Aconiti Lateralis Radix Praeparata-Cinnamomi Cortex Combination Alleviates Slow Transit Constipation due to Yang Deficiency in Rats via Regulating VIP/cAMP/PKA/AQP Pathway in Colon
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 29, Issue 21, Pages: 103-113(2023)
- 作者机构：
  
  1.贵州中医药大学,贵阳 550025
  2.贵州中医药大学第二附属医院,贵阳 550025
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20231212
  CLC： R284;R285;R289;R287;R22;R2-031;R33;R24
- Published：05 November 2023，
  
  Published Online：22 September 2023，
  
  Received：25 May 2023，
- 稿件说明：
扫描看全文
赵罗娜,杨元凤,黎豫川等.附子-肉桂调控结肠VIP/cAMP/PKA/AQP通路改善大鼠阳虚型慢传输型便秘[J].中国实验方剂学杂志,2023,29(21):103-113.

ZHAO Luona,YANG Yuanfeng,LI Yuchuan,et al.Aconiti Lateralis Radix Praeparata-Cinnamomi Cortex Combination Alleviates Slow Transit Constipation due to Yang Deficiency in Rats via Regulating VIP/cAMP/PKA/AQP Pathway in Colon[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(21):103-113.
赵罗娜,杨元凤,黎豫川等.附子-肉桂调控结肠VIP/cAMP/PKA/AQP通路改善大鼠阳虚型慢传输型便秘[J].中国实验方剂学杂志,2023,29(21):103-113. DOI： 10.13422/j.cnki.syfjx.20231212.

ZHAO Luona,YANG Yuanfeng,LI Yuchuan,et al.Aconiti Lateralis Radix Praeparata-Cinnamomi Cortex Combination Alleviates Slow Transit Constipation due to Yang Deficiency in Rats via Regulating VIP/cAMP/PKA/AQP Pathway in Colon[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(21):103-113. DOI： 10.13422/j.cnki.syfjx.20231212.

摘要

目的

通过血管活性肠肽（VIP）/环状磷酸腺苷（cAMP）/蛋白激酶A（PKA）/水通道蛋白（AQP）通路探究附子-肉桂对阳虚型慢传输便秘（STC）大鼠肠道功能的影响及作用机制。

方法

将SD大鼠随机分为6组（

=6），包括正常组、模型组、附子-肉桂高剂量组、附子-肉桂中剂量组、附子-肉桂低剂量组及普芦卡必利组。除正常组以外，其余组均采用盐酸洛哌丁胺联合冰水灌胃造模法，建立阳虚型STC大鼠模型。记录各组大鼠首次黑便排出时间，粪便数量，粪便含水量，肠道推进率及粪便性状评分。通过苏木素-伊红（HE）染色观察结肠组织病理形态，阿利新蓝-过碘酸-雪夫（AB-PAS）染色观察结肠黏液分泌情况。酶联免疫吸附测定法（ELISA）检测血清中VIP的水平。实时荧光定量聚合酶链式反应（Real-time PCR）检测结肠组织中AQPs mRNA表达水平，免疫组化检测结肠和肾组织中AQPs表达，蛋白免疫印迹法（Western blot）检测结肠组织中cAMP、PKA及VIP的蛋白表达水平。

结果

与空白组大鼠相比，模型组大鼠首粒黑便排出时间显著加长，粪便粒数及含水量显著减少，粪便性状评分及肠道推进率显著降低（

0.01）；大鼠肠道病变重，黏液分泌减少；血清中VIP含量显著减少，结肠和肾组织中水通道蛋白1（AQP1）表达水平显著增加，水通道蛋白3（AQP3）及水通道蛋白（AQP9）表达水平显著减少（

0.01）；结肠组织中cAMP、PKA及VIP表达降低。与模型组大鼠比较，高剂量附子-肉桂组大鼠首粒黑便排出时间缩短，粪便粒数及含水量增加，粪便性状评分及肠道推进率提高；附子-肉桂高剂量组，大鼠肠道病变减轻，黏液分泌增加；血清中VIP含量增加（

0.01），结肠和肾组织中AQP1表达水平明显降低，AQP3及AQP9表达水平明显增加（

0.05，

0.01）。附子-肉桂高剂量组大鼠结肠组织中cAMP、PKA及VIP表达也显著增加（

0.01）。

结论

附子-肉桂高剂量组可以通过上调VIP/cAMP/PKA/AQP通路提高肠道动力，平衡肠道水液代谢，从而改善阳虚型慢传输便秘大鼠的便秘症状。

Abstract

Objective

To investigate the effect and mechanism of Aconiti Lateralis Radix Praeparata-Cinnamomi Cortex in regulating the intestinal function in the rat model of slow transit constipation （STC） due to yang deficiency via the vasoactive intestinal peptide （VIP）/cathelicidin antimicrobial peptide （cAMP）/protein kinase A （PKA）/aquaporin （AQP） pathway.

Method

SD rats were randomized into 6 groups （

=6）， including a control group， a model group， high-， medium-， and low-dose Aconiti Lateralis Radix Praeparata-Cinnamomi Cortex groups， and a prucalopride group. Other groups except the control group were treated with loperamide hydrochloride combined with ice water by gavage for the modeling of STC due to yang deficiency. The number of fecal pellets， time to the first black stool defecation， fecal water content， intestinal propulsion rate， and score of fecal properties were recorded in each group. At the end of the treatment， the colon was stained with hematoxylin-eosin （HE） to reveal the histopathological changes and Alcian blue/periodic acid-Schiff （AB-PAS） to reveal the secretion of colonic mucus. The enzyme-linked immunosorbent assay （ELISA） was employed to measure the level of VIP in the serum. The mRNA level of AQP in the colon was measured by polymerase chain reaction （Real-time PCR）. Immunohistochemical staining was performed to observe the expression of AQPs in the colon and kidney tissues. Western blot was performed to determine the protein levels of cAMP， PKA， and VIP in the colon tissue.

Result

Compared with the control group

the model group had longer time to the first black stool defecation

reduced fecal pellets and water content

reduced Bristol Stool Form Scale score and intestinal propulsion rate

and constipation aggravated（

0.01）. Moreover

increased the intestinal lesions

reduced the mucus secretion

reduce the serum VIP level

up-regulated the expression levels of AQP1 in the colon and kidney tissues

inhibited the expression of AQP3 and AQP9（

0.01）.

and down-regulated the protein levels of cAMP

PKA

and VIP in the colon tissue. Compared with the model group， the high-dose Aconiti Lateralis Radix Praeparata-Cinnamomi Cortex group had shortened time to the first black stool defecation， increased fecal pellets and water content， increased Bristol Stool Form Scale score and intestinal propulsion rate， and alleviated constipation symptoms. Moreover， high-dose Aconiti Lateralis Radix Praeparata-Cinnamomi Cortex reduced the intestinal lesions， increased the mucus secretion， elevated the serum VIP level（

0.01）.， down-regulated the expression levels of AQP1 in the colon and kidney tissues， promoted the expression of AQP3 and AQP9（

0.05，

0.01）， and up-regulated the protein levels of cAMP， PKA， and VIP in the colon tissue. The medium- and low-dose groups had weaker effect than the high-dose group（

0.01）.

Conclusion

High-dose Aconiti Lateralis Radix Praeparata-Cinnamomi Cortex can improve the intestinal motility and balance the intestinal water and fluid metabolism by up-regulating the VIP/cAMP/PKA/AQP pathway， thereby mitigating the constipation symptoms in the rat model of slow transit constipation due to yang deficiency.

关键词

阳虚型慢传输便秘附子-肉桂血管活性肠肽（VIP）/水通道蛋白VIP/环状磷酸腺苷（cAMP）/蛋白激酶A（PKA）/水通道蛋白（AQP）通路

Keywords

slow transit constipation due to yang deficiencyAconiti Lateralis Radix Praeparata-Cinnamomi Cortexvasoactive intestinal peptideaquaporinsvasoactive intestinal peptide （VIP）/cathelicidin antimicrobial peptide （cAMP）/protein kinase A （PKA）/aquaporin （AQP） pathway

references

SUN Y，YAN C，JIN S，et al.Curative effect and mechanism of guiren runchang granules on morphine-induced slow transit constipation in mice［J］.Evid Based Complement Alternat Med，2020，2020：5493192.

TILLOU J，POYLIN V.Functional disorders： Slow-transit constipation［J］.Clin Colon Rectal Surg，2017，30（1）：76-86.

王盼盼，张小元.基于开阖枢理论探讨脾肾阳虚型便秘的辨治［J］.中国民间疗法，2022，30（19）：29-31.

王志荣.附子丁香散加味穴位贴敷治疗阳虚型STC的临床观察和实验研究［D］.济南：山东中医药大学，2018.

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.

BAINS M，LANEY C，WOLFE A E，et al.Vasoactive intestinal peptide deficiency is associated with altered gut microbiota communities in male and female C57BL/6 mice［J］.Front Microbiol，2019，10：2689.

叶俊雯.建中活血行气汤对慢传输型便秘大鼠的疗效及作用机制研究［D］.重庆：西南医科大学，2018.

IKARASHI N，MOCHIDUKI T，TAKASAKI A，et al.A mechanism by which the osmotic laxative magnesium sulphate increases the intestinal aquaporin 3 expression in HT-29 cells［J］.Life Sci，2011，88（3/4）：194-200.

杨清瑞，胡泽玉，雷玉玉，等.基于5-羟色胺信号通路探讨中医药辨证治疗腹泻型肠易激综合征的研究进展［J］.中国实验方剂学杂志，2023，doi：10.13422/j.cnki.syfjx.20232291http://dx.doi.org/10.13422/j.cnki.syfjx.20232291.

周永学，王郁金，闫曙光，等.硝菔通结方对功能性便秘大鼠结肠组织中VIP-cAMP-PKA-AQP3信号通路的影响［J］.中国实验方剂学杂志，2016，22（24）：99-104.

李智辉.附子配伍肉桂对肾阳虚小鼠治疗的实验研究［D］.昆明：云南中医学院，2008.

张远哲，黎豫川，赵罗娜.附子-肉桂对慢传输型便秘大鼠5-HT信号通路及c-Kit表达的影响［J］.世界科学技术—中医药现代化，2023，25（3）：1111-1121.

范明明，林伟，韩海瑞，等.慢传输型便秘动物模型构建方法的研究进展［J］.广西医学，2022，44（14）：1657-1660.

JARUVONGVANICH V，PATCHARATRAKUL T，GONLACHANVIT S.Prediction of delayed colonic transit using bristol stool form and stool frequency in eastern constipated patients：A difference from the west［J］.J Neurogastroenterol Motil，2017，23（4）：561-568.

王亚文.桑叶提取物对豚鼠离体肠肌及洛哌丁胺所致便秘大鼠肠道功能的影响［D］.石家庄：河北医科大学，2021.

邓振旭，楚德昌.消化道组织块黏液细胞的组织化学染色［J］.生物技术，2007，17（6）：41-43.

ZHANG D，ZHU A.Pathogenesis and treatment of slow transit constipation［J］.Chin J Gastrointest Surg，2016，19（12）：1447-1450.

Guidelines-Rome Ⅲ Diagnostic Criteria for Functional Gastrointestinal Disorders［J］.J Gastrointestin Liver Dis，2006，15（3）：307-312.

MUGIE S M，BENNINGA M A，DI LORENZO C.Epidemiology of constipation in children and adults：A systematic review［J］.Best Pract Res Clin Gastroenterol，2011，25（1）：3-18.

BHARUCHA A E，PEMBERTON J H，LOCKE G R.American gastroenterological association technical review on constipation［J］.Gastroenterology，2013，144（1）：218-238.

李硕，孙悦阳，樊景春，等.基于CiteSpace的中医药治疗便秘的知识图谱可视化分析［J］.中国实验方剂学杂志，2023，29（4）：167-175.

顾尽晖，史仁杰.慢性功能性便秘从肾阳虚论治［J］.辽宁中医药大学学报，2010，12（10）：99-101.

刘莹，侯晓华，宋军.消除对肠杯状细胞的“刻板印象”——肠杯状细胞的研究进展［J］.临床消化病杂志，2023，35（2）：159-163.

程晓雯，林中，郑清华，等.重症急性胰腺炎伴胃肠动力障碍大鼠结肠黏膜下血管活性肠肽神经元的变化［J］.南京医科大学学报：自然科学版，2012，32（2）：183-187.

武云朋.舒肝解郁胶囊对功能性便秘大鼠胃肠动力学及肠神经递质VIP、cAMP、AQP3影响机制的研究［D］.呼和浩特：内蒙古医科大学，2021.

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.

HE J，YANG B.Aquaporins in renal diseases［J］.Int J Mol Sci，2019，20（2）：366.

马雪巍，刘传佳，唐学贵.增液汤对慢传输型便秘大鼠结肠AQP9的影响及血清中5-HT的表达变化［J］.中华中医药学刊，2020，38（4）：125-129，后插11.

杜丽娟，占煜，吴至久，等.大黄对便秘大鼠肠动力及结肠水通道蛋白3表达的调节作用［J］.中华中医药学刊，2017，35（4）：873-875.

王沐晨，单思，刘红宁.水通道蛋白与“肾主水”理论的关系探析［J］.中国实验方剂学杂志，2022，28（18）：205-212.

梁星琛，周永学，张小波.基于“肾主水司二便”探讨硝菔通结方对功能性便秘大鼠水通道蛋白表达的影响［J］.中华中医药学刊，2018，36（1）：57-60.

刘娇，陈明，陈鑫，等.从水通道蛋白诠释“肾主水”的现代科学内涵［J］.山东中医杂志，2021，40（8）：884-889.

赵兵，孔鹏飞，吴至久，等.AQP1在慢传输型便秘发病过程中的作用机制探讨［J］.川北医学院学报，2014，29（6）：527-530.

LIU W，ZHI A.The potential of quercetin to protect against loperamide-induced constipation in rats［J］.Food Sci Nutr，2021，9（6）：3297-3307.

CONG L，DUAN L W，SU W P，et al.Efficacy of high specific volume polysaccharide-A new type of dietary fiber-on molecular mechanism of intestinal water metabolism in rats with constipation［J］.Med Sci Monit，2019，25：5028-5035.

KON R，IKARASHI N，HAYAKAWA A，et al.Morphine-induced constipation develops with increased aquaporin-3 expression in the colon via increased serotonin secretion［J］.Toxicol Sci，2015，145（2）：337-347.

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，2021，15：1971-1980.

ZHAN Y，TANG X，XU H，et al.Maren Pills improve constipation via regulating AQP3 and NF-κB Signaling Pathway in Slow Transit Constipation In Vitro and In Vivo［J］.Evid Based Complement Alternat Med，2020，2020：9837384.

储正达，蔡彬.肠道水通道蛋白与便秘的相互关系［J］.现代消化及介入诊疗，2023，28（1）：117-121.

ZHU C，CHEN Z，JIANG Z.Expression， distribution and role of aquaporin water channels in human and animal stomach and intestines［J］.Int J Mol Sci，2016，17（9）：1399.

杨会锋，袁维堂.水通道蛋白3和水通道蛋白9在结肠黏膜的表达及意义［J］.医药论坛杂志，2007，28（6）：12-14，封4.

EL HALAWANI M E ，PITTS G R，SUN S，et al.Active immunization against vasoactive intestinal peptide prevents photo-induced prolactin secretion in turkeys［J］.Gen Comp Endocrinol，1996，104（1）：76-83.

HALIMI F，PIOT O，GUIZE L，et al.Electrophysiological effects of vasoactive intestinal peptide in rabbit atrium： A modulation of acetylcholine activity［J］.J Mol Cell Cardiol，1997，29（1）：37-44.

丛丽敏.益生菌联合膳食纤维改善便秘［D］.大连：大连医科大学，2016.

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Anti-swelling and Analgesic Mechanism of Jianpi Tongluo Prescription from MAPKs Signaling Pathway

Relationship Between Aquaporins and "Kidney Governing Water" Theory

Mechanism of Xiao Qinglongtang Regulating Lung Water Transport-related Proteins Based on Theory of Lung Controlling Water Movement

Mechanism of Chinese Medicine Against Sjögren Syndrome： A Review

Diuretic Activity and Mechanism of

Related Author

FANG-LUO Changting

LIU Chunfang

LIN Na

WANG Hongjie

XU Tengteng

YANG Chao

HU Zhixing

WANG Xiaoxiao

Related Institution

Institute of Chinese Materia Medica， China Academy of Chinese Medicine Sciences

Research Center for Differentiation and Development of Basic Theory of Chinese Medicine，Jiangxi University of Chinese Medicine

Jiangxi Key Laboratory of Etiology and Biology of Chinese Medicine

School of Traditional Chinese Medicine， Beijing University of Chinese Medicine

Dongzhimen Hospital，Beijing University of Chinese Medicine

Postal code：100700
Tel：010-84076882 Email：syfjx_2010@188.com
Technical support is provided by Beijing Founder electronics co., LTD 京ICP备11006657号-10 京公网安备11010802024621
It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.

⁰