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1.成都中医药大学,成都 611137
2.四川省中医药科学院 中医研究所,成都 610031
3.华润(雅安)三九药业集团,四川 雅安 625000
刘芳,博士,高级实验师,从事中药炮制与中药制剂研究,E-mail:bittercaffe1@126.com
刘畅,硕士,从事中药炮制与中药制剂研究,E-mail:Liuchang2548@126.com
收稿日期:2022-10-13,
网络出版日期:2022-12-14,
纸质出版日期:2023-07-20
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刘芳,张芮苑,刘畅等.发汗前后厚朴对IBS-C大鼠肠道菌群分布及5-HT、SP水平的影响[J].中国实验方剂学杂志,2023,29(14):141-149.
LIU Fang,ZHANG Ruiyuan,LIU Chang,et al.Effect of Magnolia officinalis Cortex Before and After Sweating on Distribution of Intestinal Flora and Levels of 5-HT and SP in IBS-C Rats[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(14):141-149.
刘芳,张芮苑,刘畅等.发汗前后厚朴对IBS-C大鼠肠道菌群分布及5-HT、SP水平的影响[J].中国实验方剂学杂志,2023,29(14):141-149. DOI: 10.13422/j.cnki.syfjx.20230346.
LIU Fang,ZHANG Ruiyuan,LIU Chang,et al.Effect of Magnolia officinalis Cortex Before and After Sweating on Distribution of Intestinal Flora and Levels of 5-HT and SP in IBS-C Rats[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(14):141-149. DOI: 10.13422/j.cnki.syfjx.20230346.
目的
2
研究发汗前后厚朴对便秘型肠易激综合征(IBS-C)大鼠的作用机制。
方法
2
采用冰水灌胃法建立IBS-C大鼠模型。将大鼠随机分为空白组、模型组、莫沙必利组(1 mg·kg
-1
)、未发汗厚朴组(以下简称“厚朴组”,10 g·kg
-1
)及发汗厚朴组(10 g·kg
-1
)。观察大鼠体质量、粪便含水量、粪便数量变化;采用16S rRNA测序检测大鼠粪便肠道菌群的结构变化;采用酶联免疫吸附测定法(ELISA)测定大鼠结肠组织中5-羟色胺(5-HT)及P物质(SP)水平。
结果
2
与模型组比较,给药后莫沙必利组、厚朴组、发汗厚朴组的粪便含水量和粪便数量明显升高(
P
<
0.05)。16S rRNA测序结果显示,菌群门水平丰度前4种分别为厚壁菌门、拟杆菌门、螺旋体门、变形菌门,与空白组比较,模型组厚壁菌门相对丰度明显降低(
P
<
0.05),螺旋体门相对丰度明显增加(
P
<
0.05);与模型组比较,厚朴组及发汗厚朴组的厚壁菌门、螺旋体门比例趋近于空白组,且发汗厚朴组螺旋体门相对丰度低于厚朴组;在科水平上,菌群丰度前4种分别为乳酸菌科、S24_7、瘤胃球菌科、拟杆菌科,与空白组比较,模型组的乳酸菌科相对丰度明显降低(
P
<
0.05),模型大鼠给药后厚朴组和发汗厚朴组的乳酸菌科相对丰度明显提高(
P
<
0.05),且两组大鼠菌群结构趋近于空白组;在属水平上,菌群丰度前4种分别为乳酸菌属、Unspecified_
S
24_7、拟杆菌属、密螺旋体属,与空白组比较,模型组的乳酸菌属相对丰度明显降低(
P
<
0.05),密螺旋体属的相对丰度则明显增加(
P
<
0.05);与模型组比较,厚朴组与发汗厚朴组乳酸菌属和密螺旋体属菌群结构比例趋近于空白组,说明发汗前后厚朴均可恢复IBS-C大鼠肠道菌群结构。药理实验结果显示,与模型组比较,莫沙必利组的5-HT含量明显降低(
P
<
0.05),厚朴组与发汗厚朴组的5-HT和SP含量均显著升高(
P
<
0.01),且发汗厚朴组高于厚朴组。
结论
2
发汗前后厚朴均可通过改善肠道菌群结构、调节5-HT和SP含量治疗IBS-C。
Objective
2
To investigate the mechanism of
Magnolia officinalis
cortex for constipation-type irritable bowel syndrome(IBS-C) rats before and after sweating.
Method
2
IBS-C rat model was established by gavage of ice water, and rats were randomly divided into the blank group, model group, mosapride group(1 mg·kg
-1
),
M. officinalis
cortex group(10 g·kg
-1
) and sweated
M. officinalis
cortex group(10 g·kg
-1
). The changes of body weight, fecal number and fecal water content of rats were observed, 16S rRNA sequencing was used to detect the structural changes of fecal intestinal flora in rats, the levels of 5-hydroxytryptamine(5-HT) and substance P(SP) in colonic tissues of rats were determined by enzyme-linked immunosorbent assay(ELISA).
Result
2
Compared with the model group, the fecal water content and fecal number of mosapride group,
M. officinalis
cortex group and sweated
M. officinalis
cortex group were significantly increased(
P
<
0.05). At the phylum level, the top four species of flora abundance were Firmicutes, Bacteroidetes, Spirochaetes and Proteobacteria. Compared with the blank group, the proportion of Firmicutes in the model group was significantly reduced(
P
<
0.05), while the proportion of Spirochaetes was significantly increased(
P
<
0.05). Compared with the model group, the proportion of Firmicutes and Spirochaetes in
M. officinalis
cortex group and sweated
M. officinalis
cortex group tended to be similar to that in the blank group, and the proportion of Spirochaetes in sweated
M. officinalis
cortex group was lower than that of
M. officinalis
cortex group. At the family level, the top four species of flora abundance were Lactobacillaceae, S24_7, Ruminococcaceae, Bacteroidaceae, compared with the blank group, the proportion of Lactobacillaceae in the model group decreased significantly(
P
<
0.05), and its proportion in the
M. officinalis
cortex group and sweated
M. officinalis
cortex group increased significantly after administration(
P
<
0.05), and the flora structure of the two groups tended to be similar to that of the blank group. At the genus level, the top four species of flora abundance were
Lactobacillus
, Unspecified_
S
24_7,
Bacteroides
and
Treponema
. Compared with the blank group, the proportion of
Lactobacillus
in the model group decreased significantly(
P
<
0.05), while the proportion of
Treponema
increased significantly(
P
<
0.05). Compared with the model group, ratio of bacterial structure of
Lactobacillus
and
Treponema
in the
M. officinalis
cortex group and sweated
M. officinalis
cortex group tended to be similar to those in the blank group, indicating that
M. officinalis
cortex could restore the intestinal microbial structure of IBS-C rats before and after sweating. Compared with the model group, the 5-HT content in mosapride group was significantly reduced(
P
<
0.05), the contents of 5-HT and SP in the
M. officinalis
cortex group and sweated
M. officinalis
cortex group were significantly increased(
P
<
0.01), and the sweated
M. officinalis
cortex group was higher than the
M. officinalis
cortex group.
Conclusion
2
M. officinalis
cortex can play a therapeutic role on IBS-C rats by regulating 5-HT pathway and intestinal flora structure before and after sweating.
郑超伟 , 李桂贤 , 李敏 , 等 . 加味柴芍六君颗粒对便秘型肠易激综合征模型大鼠结肠组织5-HT的影响 [J]. 实用中医药杂志 , 2019 , 35 ( 4 ): 386 - 388 .
CAMILLERI M , LASCH K , ZHOU W . The confluence of increased permeability,inflammation,and pain in irritable bowel syndrome [J]. Am J Physiol Gastrointest Liver Physiol , 2012 , 303 ( 7 ): G775 - G785 .
OCCHIPINTI K , SMITH J W . Irritable bowel syndrome:A review and update [J]. Clin Colon Rectal Surg , 2012 , 25 ( 1 ): 46 - 52 .
GILET M , EUTAMENE H , HAN H , et al . Influence of a new 5-HT4 receptor partial agonist,YKP10811,on visceral hypersensitivity in rats triggered by stress and inflammation [J]. Neurogastroenterol Motil , 2014 , 26 ( 12 ): 1761 - 1770 .
郝强 , 郝春华 , 庄贺 . 电针天枢穴和太冲穴对便秘型肠易激综合征模型小鼠5-羟色胺信号系统的影响 [J]. 山东中医药大学学报 , 2020 , 44 ( 1 ): 69 - 72 .
SAHA L . Irritable bowel syndrome:Pathogenesis,diagnosis,treatment,and evidence-based medicine [J]. World J Gastroenterol , 2014 , 20 ( 22 ): 6759 - 6773 .
李潇潇 , 卢圣锋 , 朱冰梅 , 等 . 5-HT与肠道菌群及其在肠-脑相关疾病中的作用研究进展 [J]. 中国康复医学杂志 , 2019 , 34 ( 1 ): 116 - 119 .
谷诺诺 . 从神经调控与肠道微生物的相关性分析通腑降浊方治疗IBS-C模型大鼠作用机制的实验研究 [D]. 石家庄 : 河北医科大学 , 2018 .
国家药典委员会 . 中华人民共和国药典:一部 [M]. 北京 : 中国医药科技出版社 , 2020 : 263 .
郭雅婧 . 谢晶日教授治疗便秘型肠易激综合征的临床经验总结 [D]. 哈尔滨 : 黑龙江中医药大学 , 2021 .
王潇 , 王婷 , 张晨 , 等 . 基于代谢组学研究厚朴“发汗”前后对胃肠动力障碍大鼠的影响 [J]. 中国中药杂志 , 2019 , 44 ( 6 ): 1170 - 1178 .
WANG X , ZHANG C , ZHENG M Y , et al . Metabolomics analysis of L -arginine induced gastrointestinal motility disorder in rats using UPLC-MS after magnolol treatment [J]. Front Pharmacol , 2019 , 10 : 183 .
刘畅 , 王潇 , 刘芳 , 等 . 基于多指标质量差异关键属性优化厚朴产地加工“发汗”工艺 [J]. 中草药 , 2021 , 52 ( 3 ): 677 - 684 .
刘永 , 谷诺诺 , 王凯星 , 等 . 通腑降浊方对便秘型肠易激综合征大鼠粪便菌群结构的影响 [J]. 广州中医药大学学报 , 2020 , 37 ( 6 ): 1127 - 1133 .
万丹 , 梁雪娟 , 何丹 , 等 . 整合网络药理学和生物信息学分析便可通片抗便秘性肠易激综合征分子机制 [J]. 中国实验方剂学杂志 , 2021 , 27 ( 20 ): 152 - 161 .
孙建梅 , 顾立梅 , 李慧 , 等 . 运脾柔肝方对便秘型肠易激综合征大鼠5-HT、VIP及SP影响的实验研究 [J]. 南京中医药大学学报 , 2017 , 33 ( 5 ): 524 - 528 .
王潇 , 王婷 , 张晨 , 等 . 人工神经网络优化厚朴提取工艺及其“发汗”前后的含量测定 [J]. 中华中医药学刊 , 2019 , 37 ( 12 ): 2978 - 2982 .
王丁 , 张韶峰 , 姜雪 , 等 . 益元起痿丸治疗糖尿病性勃起功能障碍大鼠作用机制 [J]. 中国实验方剂学杂志 , 2022 , 28 ( 18 ): 77 - 84 .
CALLAHAN B J , MCMURDIE P J , ROSEN M J , et al . DADA2:High-resolution sample inference from Illumina amplicon data [J]. Nat Methods , 2016 , 13 ( 7 ): 581 - 583 .
LOVE M I , HUBER W , ANDERS S . Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2 [J]. Genome Biol , 2014 , 15 ( 12 ): 550 .
MIWA J , ECHIZEN H , MATSUEDA K , et al . Patients with constipation-predominant irritable bowel syndrome (IBS) may have elevated serotonin concentrations in colonic mucosa as compared with diarrhea-predominant patients and subjects with normal bowel habits [J]. Digestion , 2000 , 63 ( 3 ): 188 - 194 .
崔艳 , 张亚伟 , 肖林 , 等 . 肠道有益菌及代谢产物对肠道和人体的影响 [J]. 生物产业技术 , 2017 , 9 ( 1 ): 91 - 94 .
LUO H , WU H , YU X , et al . A review of the phytochemistry and pharmacological activities of Magnoliae officinalis cortex [J]. J Ethnopharmacol , 2019 , 236 : 412 - 442 .
师哲 , 周蒙恩 , 张克慧 , 等 . 加味芪榔方治疗药物依赖性气阴两虚型便秘患者的疗效及对外周血MTL,VIP,5-HT和5-HT4R的影响 [J]. 中国实验方剂学杂志 , 2021 , 27 ( 15 ): 51 - 56 .
EL-SALHY M , WENDELBO I , GUNDERSEN D . Serotonin and serotonin transporter in the rectum of patients with irritable bowel disease [J]. Mol Med Rep , 2013 , 8 ( 2 ): 451 - 455 .
LAYUNTA E , LATORRE E , FORCÉN R , et al . NOD1 downregulates intestinal serotonin transporter and interacts with other pattern recognition receptors [J]. J Cell Physiol , 2018 , 233 ( 5 ): 4183 - 4193 .
REIGSTAD C S , LINDEN D R , SZURSZEWSKI J H , et al . Correlated gene expression encoding serotonin(5-HT) receptor 4 and 5-HT transporter in proximal colonic segments of mice across different colonization states and sexes [J]. Neurogastroenterol Motil , 2016 , 28 ( 9 ): 1443 - 1448 .
杨倩 , 邱贝 , 马赟 , 等 . 便秘型肠易激综合征大鼠肠组织中5-HT的实验研究 [J]. 中医临床研究 , 2014 , 6 ( 30 ): 35 - 37 .
裴丽霞 . 基于5-HT信号系统探讨电针治疗便秘型肠易激综合征的机制 [D]. 南京 : 南京中医药大学 , 2017 .
KOH A , VADDER F D , KOVATCHEVA-DATCHARY P , et al . From dietary fiber to host physiology:Short-chain fatty acids as key bacterial metabolites [J]. Cell , 2016 , 165 ( 6 ): 1332 - 1345 .
KAMADA N , SEO S U , CHEN G Y , et al . Role of the gut microbiota in immunity and inflammatory disease [J]. Nat Rev Immunol , 2013 , 13 ( 5 ): 321 - 335 .
REIGSTAD C S , SALMONSON C E , RAINEY J F , et al . Gut microbes promote colonic serotonin production through an effect of short-chain fatty acids on enterochromaffin cells [J]. FASEB J , 2015 , 29 ( 4 ): 1395 - 403 .
赵倩倩 , 倪喆鑫 , 毕艳丽 , 等 . 化瘀解毒方改善子宫内膜异位症小鼠肠道菌群及粪便代谢组 [J]. 中国实验方剂学杂志 , 2021 , 27 ( 9 ): 202 - 214 .
张薇 , 葛文静 , 王慧森 , 等 . 痛泻要方加减引经药防风对肠易激综合征大鼠水液代谢和5-HT系统的调控作用 [J]. 中国实验方剂学杂志 , 2020 , 26 ( 11 ): 56 - 62 .
O'MAHONY S M , CLARKE G , BORRE Y E , et al . Serotonin,tryptophan metabolism and the brain-gut-microbiome axis [J]. Behav Brain Res , 2015 , 277 : 32 - 48 .
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