Effects of moxibustion on intestinal barrier function and TLR4/NF-κB p65 signaling pathway in  obese rats

CHEN Xia; ZHANG Yanji; HUANG Wei; ZHANG Yingrong; KONG Weiqing; ZHOU Zhongyu

doi:10.13703/j.0255-2930.20230529-k0003

您当前的位置：

首页 >

文章列表页 >

Effects of moxibustion on intestinal barrier function and TLR4/NF-κB p65 signaling pathway in obese rats

Study on Mechanism | 更新时间：2024-06-03

- Effects of moxibustion on intestinal barrier function and TLR4/NF-κB p65 signaling pathway in obese rats
- Chinese Acupuncture & Moxibustion Vol. 44, Issue 4, Pages: 449-454(2024)
- 作者机构：
  
  1.湖北中医药大学针灸骨伤学院，武汉430065
  2.湖北省中医院，武汉430061
  3.湖北中医药大学附属医院，武汉430061
- 作者简介：
- 基金信息：
- DOI：10.13703/j.0255-2930.20230529-k0003
  CLC：
- Received：29 May 2023，
  
  Published：12 April 2024
- 稿件说明：
移动端阅览
陈霞,张艳佶,黄伟,等.艾灸对肥胖大鼠肠道屏障功能及TLR4/NF-κB p65信号通路的影响[J].中国针灸,2024,44(4):449-454.

CHEN Xia,ZHANG Yanji,HUANG Wei,et al.Effects of moxibustion on intestinal barrier function and TLR4/NF-κB p65 signaling pathway in obese rats[J].Chinese Acupuncture & Moxibustion,2024,44(04):449-454.
陈霞,张艳佶,黄伟,等.艾灸对肥胖大鼠肠道屏障功能及TLR4/NF-κB p65信号通路的影响[J].中国针灸,2024,44(4):449-454. DOI： 10.13703/j.0255-2930.20230529-k0003.

CHEN Xia,ZHANG Yanji,HUANG Wei,et al.Effects of moxibustion on intestinal barrier function and TLR4/NF-κB p65 signaling pathway in obese rats[J].Chinese Acupuncture & Moxibustion,2024,44(04):449-454. DOI： 10.13703/j.0255-2930.20230529-k0003.

摘要

目的

观察艾灸对肥胖大鼠肠道屏障功能及Toll样受体4（TLR4）/核转录因子κB p65（NF-κB p65）信号通路的调控作用，探究艾灸干预肥胖的作用机制。

方法

将55只SPF级7周龄雄性Wistar大鼠随机分为正常组（10只）和造模组（45只），造模组采用高脂饮食喂养建立肥胖大鼠模型。造模成功的30只大鼠随机分为模型组、艾灸组、安慰对照组，每组10只。艾灸组在距离“中脘”3~5 cm处施灸，温度保持在（46±1）℃；安慰对照组在距离“中脘”8~10 cm处施灸，温度保持在（38±1）℃。均每天干预1次，共干预8周。观察各组大鼠干预前后体质量、摄食量；ELISA法测定各组大鼠血清总胆固醇（TC）、三酰甘油（TG）、脂多糖（LPS）水平，计算胰岛素抵抗指数（HOMA-IR）；HE染色法观察各组大鼠结肠组织形态；实时荧光定量PCR法检测各组大鼠结肠闭锁连接蛋白-1（ZO-1）、咬合蛋白（Occludin）、紧密连接蛋白-1（Claudin-1）、TLR4、NF-κB p65 mRNA表达；Western blot法检测各组大鼠结肠ZO-1、Occludin、Claudin-1、TLR4和NF-κB p65蛋白表达。

结果

与正常组比较，模型组大鼠体质量、摄食量、HOMA-IR及血清TC、TG、LPS水平升高（

0.01）；与模型组和安慰对照组比较，艾灸组大鼠体质量、摄食量、HOMA-IR及血清TC、TG、LPS水平降低（

0.01，

0.05）。与正常组比较，模型组大鼠结肠组织黏膜大量上皮细胞损伤脱落，间质内可见明显炎性细胞浸润。与模型组比较，艾灸组大鼠结肠组织损伤有不同程度恢复，间质内可见少量炎性细胞浸润；安慰对照组大鼠结肠组织上皮损伤有少许恢复，间质内可见炎性细胞浸润。与正常组比较，模型组大鼠结肠ZO-1、Occludin、Claudin-1 mRNA和蛋白表达降低（

0.01），结肠TLR4、NF-κB p65 mRNA和蛋白表达升高（

0.01）；与模型组和安慰对照组比较，艾灸组大鼠结肠ZO-1、Occludin、Claudin-1 mRNA和蛋白表达升高（

0.01，

0.05），结肠TLR4、NF-κB p65 mRNA和蛋白表达降低（

0.01）。

结论

艾灸可以降低肥胖大鼠体质量、摄食量，调节肥胖大鼠血脂，改善胰岛素抵抗，可能与改善肠道屏障功能、调节肠道TLR4/NF-κB p65信号通路来减轻炎性反应有关。

Abstract

Objective

To observe the effects of moxibustion on intestinal barrier function and Toll-like receptor 4 (TLR4)/nuclear factor-κB p65 (NF-κB p65) signaling pathway in obese rats and explore the mechanism of moxibustion in the intervention of obesity.

Methods

Fifty-five Wistar rats of SPF grade were randomly divided into a normal group (10 rats) and a modeling group (45 rats). In the modeling group

the obesity model was established by feeding high-fat diet. Thirty successfully-modeled rats were randomized into a model group

a moxibustion group

and a placebo-control group

with 10 rats in each one. In the moxibustion group

moxibustion was applied at the site 3 cm to 5 cm far from the surface of "Zhongwan" (CV 12)

with the temperature maintained at (46±1 ) ℃. In the placebo-control group

moxibustion was applied at the site 8 cm to 10 cm far from “Zhongwan” (CV 12)

with the temperature maintained at (38±1) ℃. The intervention was delivered once daily for 8 weeks in the above two groups. The body mass and food intake of the rats were observed before and after intervention in each group. Using ELISA methool

the levels of serum triacylglycerol (TG)

total cholesterol (TC) and lipopolysaccharide (LPS) were detected and the insulin resistance index (HOMA-IR) was calculated. HE staining was used to observe the morphology of colon tissue. The mRNA expression of zonula occludens-1 (ZO-1)

Occludin

Claudin-1

TLR4 and NF-κB p65 in the colon tissue was detected by quantitative real-time PCR; and the protein expression of ZO-1

Occludin

Claudin-1

TLR4 and NF-κB p65 was detected by Western blot in the rats of each group.

Results

Compared with the normal group

the body mass

food intake

the level of HOMA-IR

and the serum levels of TC

TG and LPS were increased in the rats of the model group (

0.01); those indexes in the moxibustion group were all reduced when compared with the model group and the placebo-control group respectively (

0.01

0.05). Compared with the normal group

a large number of epithelial cells in the muc

osa of colon tissue was damaged

shed

and the inflammatory cells were infiltrated obviously in the interstitium in the rats of the model group. When compared with the model group

in the moxibustion group

the damage of the colon tissue was recovered to various degrees and there were few infiltrated inflammatory cells in the interstitium

while

the epithelial injury of the colon tissue was slightly recovered and the infiltrated inflammatory cells in the interstitium were still seen in the placebo-control group. The mRNA and protein expressions of ZO-1

Occludin and Caudin-1 were decreased in the model group compared with those in the normal group (

0.01). When compared with the model group and the placebo-control group

the mRNA and protein expressions of these indexes were increased in the moxibustion group (

0.01

0.05). In the model group

the mRNA and protein expressions of TLR4 and NF-κB p65 were increased when compared with those in the normal group (

0.01)

and the mRNA and protein expressions of these indexes were reduced in the moxibustion group when compared with those in the model group and the placebo-control group (

0.01).

Conclusion

Moxibustion can reduce the body mass and food intake

regulate the blood lipid and improve insulin resistance in the rats of obesity. It may be related to alleviating inflammatory response through improving intestinal barrier function and modulating the intestinal TLR4/NF-κB p65 signaling pathway.

关键词

Keywords

references

张雪亮 . 现代肥胖症与减肥疗法 [M ] . 北京 : 北京学苑出版社 , 1990 .

Hotamisligil GS , Shargill NS , Spiegelman BM . Adipose expression of tumor necrosis factor-alpha: direct role in obesity-linked insulin resistance [J ] . Science , 1993 , 259 ( 5091 ): 87 - 91 .

李欣 , 吴伟华 . 肠道稳态与慢性低度系统性炎症 [J ] . 医学综述 , 2018 , 24 ( 7 ): 1354 - 1359 .

Ying W , Mahata S , Bandyopadhyay GK , et al . Catestatin inhibits obesity-induced macrophage infiltration and inflammation in the liver and suppresses hepatic glucose production, leading to improved insulin sensitivity [J ] . Diabetes , 2018 , 67 ( 5 ): 841 - 848 .

张金卫 , 林汉杰 , 韩凌 . 肠上皮细胞紧密连接的研究进展 [J ] . 中国医药导报 , 2015 , 12 ( 6 ): 160 - 163 .

Zihni C , Mills C , Matter K , et al . Tight junctions: from simple barriers to multifunctional molecular gates [J ] . Nat Rev Mol Cell Biol , 2016 , 17 ( 9 ): 564 - 580 .

Sumithran P , Prendergast LA , Delbridge E , et al . Long-term persistence of hormonal adaptations to weight loss [J ] . N Engl J Med , 2011 , 365 ( 17 ): 1597 - 1604 .

王琳钰 , 苏同生 , 梁莹 , 等 . 艾灸治疗肥胖的疗效特点及作用机制探析 [J ] . 辽宁中医药大学学报 , 2023 , 25 ( 8 ): 116 - 120 .

湖北省中医药学会 , 湖北省针灸学会 . 阴阳调理灸技术操作规范 [J ] . 湖北中医药大学学报 , 2022 , 24 ( 5 ): 92 - 94 .

陈霞 , 周仲瑜 . 阴阳调理灸结合五音疗法治疗痰湿质腹型肥胖42例 [J ] . 中国针灸 , 2022 , 42 ( 11 ): 1260 - 1262 .

周仲瑜 , 王佳捷 . 阴阳调理灸在更年期肥胖女性中的临床应用 [J ] . 皮肤科学通报 , 2019 , 36 ( 2 ): 225 - 230 .

张艳佶 . 穴位埋线联合阴阳调理灸改善腹型肥胖患者气虚体质的临床研究 [D ] . 武汉 : 湖北中医药大学 , 2021 .

储浩然 , 王宇 , 仝理 , 等 . 艾灸对腹泻型肠易激综合征大鼠结肠TLR4/MyD88/NF-κB信号通路的影响 [J ] . 针刺研究 , 2020 , 45 ( 8 ): 633 - 639 .

Zhang D , Ma XP , Wu HG , et al . Efficacy of herb-partitioned moxibustion at Qihai (CV 6) and bilateral Tianshu (ST 25) on colonic damage and the TLR4/NF-ÎºB signaling pathway in rats with Crohn's disease [J ] . J Tradit Chin Med , 2018 , 38 ( 2 ): 218 - 226 .

孙志 , 张中成 , 刘志诚 . 营养性肥胖动物模型的实验研究 [J ] . 中国药理学通报 , 2002 , 18 ( 4 ): 466 - 467 .

朱玉强 , 叶兰欣 , 王伊伊 , 等 . 《针灸大成》中脘穴临床应用规律探析 [J ] . 中医药临床杂志 , 2021 , 33 ( 4 ): 605 - 609 .

Su YS , Xin JJ , Yang ZK , et al . Effects of different local moxibustion-like stimuli at Zusanli (ST36) and Zhongwan (CV12) on gastric motility and its underlying receptor mechanism [J ] . Evid Based Complement Alternat Med , 2015 , 2015 : 486963 .

王桂英 , 王耀帅 , 卢开信 , 等 . 不同灸温对高脂血症小鼠胆固醇及"神阙" 穴皮肤的影响 [J ] . 中国针灸 , 2016 , 36 ( 1 ): 59 - 63 .

纵姣艳 , 王耀帅 , 王茜 , 等 . 不同灸温对高脂血症小鼠血脂和神阙穴皮肤形态及肥大细胞的影响 [J ] . 时珍国医国药 , 2017 , 28 ( 12 ): 3020 - 3022 .

Bao CH , Wu LY , Wang D , et al . Acupuncture improves the symptoms, intestinal microbiota, and inflammation of patients with mild to moderate Crohn's disease: a randomized controlled trial [J ] . EClinicalMedicine , 2022 , 45 : 101300 .

Shen L . Tight junctions on the move: molecular mechanisms for epithelial barrier regulation [J ] . Ann N Y Acad Sci , 2012 , 1258 : 9 - 18 .

Fasano A . Physiological, pathological, and therapeutic implications of zonulin-mediated intestinal barrier modulation: living life on the edge of the wall [J ] . Am J Pathol , 2008 , 173 ( 5 ): 1243 - 1252 .

Trøseid M , Nestvold TK , Rudi K , et al . Plasma lipopolysaccharide is closely associated with glycemic control and abdominal obesity: evidence from bariatric surgery [J ] . Diabetes Care , 2013 , 36 ( 11 ): 3627 - 3632 .

Allam-Ndoul B , Castonguay-Paradis S , Veilleux A . Gut microbiota and intestinal trans-epithelial permeability [J ] . Int J Mol Sci , 2020 , 21 ( 17 ): 6402 .

Monte SV , Caruana JA , Ghanim H , et al . Reduction in endotoxemia, oxidative and inflammatory stress, and insulin resistance after Roux-en-Y gastric bypass surgery in patients with morbid obesity and type 2 diabetes mellitus [J ] . Surgery , 2012 , 151 ( 4 ): 587 - 593 .

杨姝瑞 , 周钰点 , 陈瑞 , 等 . 不同腧穴配伍电针对肥胖大鼠脂代谢和肝脏Toll样受体4/核转录因子κB信号通路的影响 [J ] . 针刺研究 , 2021 , 46 ( 10 ): 845 - 850 .

Rogero MM , Calder PC . Obesity, inflammation, toll-like receptor 4 and fatty acids [J ] . Nutrients , 2018 , 10 ( 4 ): 432 .

刘畅 , 荆科 . Toll样受体阻断剂对内毒素血症小鼠肠黏膜损伤的影响 [J ] . 中国当代儿科杂志 , 2018 , 20 ( 2 ): 158 - 163 .

陈丽 , 王静芝 , 周广文 , 等 . 电针对肥胖大鼠肠道Toll样受体4和核转录因子κB的影响 [J ] . 针刺研究 , 2020 , 45 ( 7 ): 541 - 547 .

He XM , Zheng YQ , Liu SZ , et al . MiR-146a protects small intestine against ischemia/reperfusion injury by down-regulating TLR4/TRAF6/NF-κB pathway [J ] . J Cell Physiol , 2018 , 233 ( 3 ): 2476 - 2488 .

武欢 , 陈丽 , 张照庆 , 等 . 电针对肥胖胰岛素抵抗大鼠下丘脑TLR4/IκBα/NF-κB信号通路的影响 [J ] . 针刺研究 , 2021 , 46 ( 1 ): 33 - 38 .

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Mechanisms of moxibustion for delaying physical aging by regulating muscle-fat metabolism based on the theory of "Xieli Yinyang"

Electroacupuncture promotes browning of white adipose tissue by regulating mTOR/SREBP1 signaling pathway in obese mice

Effect of moxibustion-induced deqi on hippocampal autophagy function in APPswe/PSEN1dE9 transgenic Alzheimer′s disease mice

Moxibustion combined with Jianpi Wenshen formula for cancer-related anorexia of spleen-kidney yang deficiency: a randomized controlled trial

Effect of oblique needling after equal division and positioning of anatomic points on acute sacroiliac joint injury assessed with infrared thermography

Related Author

HUANG Wei

ZHOU Zhongyu

YANG Danchen

ZHANG Yuan

CHEN Fangfang

YANG Yuting

WANG Yifan

WANG Zhongyu

Related Institution

Hubei Provincial Hospital of CM

School of TCM, Ministry of Education, Beijing University of CM

School of Acupuncture-Moxibustion and Tuina, Ministry of Education, Beijing University of CM

School of Life Sciences, Ministry of Education, Beijing University of CM

Key Laboratory of Health Preservation of TCM, Ministry of Education, Beijing University of CM

⁰