柴参解郁汤对腹泻型肠易激综合征小鼠粪便胆汁酸代谢轮廓的影响

王艳天; 顾欣; 王帆; 朱梦婷; 徐坠成; 徐艺

doi:10.14148/j.issn.1672-0482.2022.0220

柴参解郁汤对腹泻型肠易激综合征小鼠粪便胆汁酸代谢轮廓的影响

doi: 10.14148/j.issn.1672-0482.2022.0220

王艳天^{1, 2,},
顾欣^{1, 2},
王帆^{1, 2},
朱梦婷^{1, 2},
徐坠成³,
徐艺^{1, 2, ,}

1.
南京中医药大学附属医院, 江苏南京 210029
2.
南京中医药大学第一临床医学院, 江苏南京 210023
3.
南京中医药大学药学院, 江苏南京 210023

基金项目:

江苏省中医药科技发展计划 RC201903

江苏省“333高层次人才培养工程”科研项目 LGY2018060

详细信息

作者简介:
王艳天，女，硕士研究生，E-mail: 20190032@njucm.edu.cn

通讯作者:
徐艺, 女，主任中医师, 主要从事脾胃病的中医药防治研究, E-mail: fsyy00638@njucm.edu.cn

中图分类号: R285.5
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- 被引次数: 0
出版历程
- 收稿日期: 2021-10-12
- 网络出版日期: 2022-03-24
- 发布日期: 2022-03-10

Effects of Chaishen Jieyu Decoction on the Metabolic Profile of Fecal Bile Acids in Mice with Diarrhea Predominant Irritable Bowel Syndrome

WANG Yan-tian^{1, 2
,},
GU Xin^{1, 2},
WANG Fan^{1, 2},
ZHU Meng-ting^{1, 2},
XU Zhui-cheng³,
XU Yi^{1, 2
, ,}

1.
The Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing 210029, China
2.
The First Clinical Medical College, Nanjing University of Chinese Medicine, Nanjing 210023, China
3.
School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing 210023, China

摘要

摘要: 目的通过研究柴参解郁汤对腹泻为主型肠易激综合征(Diarrhea predominant irritable bowel syndrome, IBS-D)模型小鼠粪便中胆汁酸代谢轮廓的影响, 探讨柴参解郁汤改善IBS-D的可能机制。方法将C57BL/6小鼠随机分为3组, 空白组、模型组、柴参解郁汤组(15 g·kg^-1)。IBS-D小鼠模型采用束缚性应激复合灌胃番泻叶造模, 造模成功后, 灌胃给予柴参解郁汤2周, 分析小鼠粪便含水量, 检测空肠病理损伤, Western blot检测空肠中COX-2、iNOS的蛋白表达, 通过检测FITC标记的右旋糖苷及紧密连接蛋白ZO-1、OCCLUDIN的mRNA表达，测定小鼠胃肠道通透性。进一步采用靶向代谢组学分析小鼠粪便胆汁酸代谢轮廓。结果与空白组相比, 模型组小鼠粪便含水量增高(P < 0.01), 给予柴参解郁汤后粪便含水量下降(P < 0.01);与空白组相比, 模型组小鼠空肠中COX-2和iNOS蛋白表达量上调(P < 0.05，P < 0.01), 而柴参解郁汤可下调空肠细胞中iNOS蛋白表达(P < 0.01);与空白组相比，模型组小鼠空肠中ZO-1、OCCLUDIN的mRNA表达量明显下降(P < 0.01)，FIFC-右旋糖苷含量下降(P < 0.01)，柴参解郁汤治疗后表达量有所回调(P < 0.05，P < 0.01)。与空白组相比, 模型组小鼠空肠绒毛大部分脱落坏死, 绒毛的固有层细胞溶解, 绒毛结构模糊不清, 给予柴参解郁汤后趋于正常。IBS-D小鼠粪便中胆汁酸组成和含量发生显著改变, 柴参解郁汤可显著回调模型组小鼠的胆汁酸代谢紊乱, 如石胆酸(LCA)、去氧胆酸(DCA)、鹅去氧胆酸(CDCA)等。结论柴参解郁汤可有效缓解IBS-D小鼠空肠组织病理改变并能改善IBS-D小鼠胆汁酸异常变化, 这可能是由于柴参解郁汤修复肝肠循环胆汁酸合成和转化途径, 增加结合型胆汁酸含量而起到保护肠道的作用。
- 肠易激综合征 /
- 柴参解郁汤 /
- 胆汁酸 /
- 肠黏膜屏障 /
- 代谢轮廓
Abstract: OBJECTIVE To investigate therapeutic effect and potential mechanism of the Chaishen Jieyu Decoction on IBS-D mice model from regulating the fecal bile acid profile. METHODS The C57BL/6 mice were randomly divided to 3 groups, normal mice, IBS-D mice and the Chaishen Jieyu Decoction intervention group. IBS-D mice were induced by the resisting immobilization stress and gavage of Folium senna. We studied the pathological changes of the mouse jejunum sections by HE, the protein expression of COX-2 and iNOS in jejunum, FITC-dextran and detected the relative mRNA expressions of main tight junctions proteins(ZO-1 & OCCLUDIN) in mice to determine the gastrointestinal permeability and analysis of fecal bile acid profile in mice using targeted metabolomics method. RESULTS Compared with the control group, the fecal water content had an uptrend in the model group, showed a decreasing trend after treatment with Chaishen Jieyu Decoction. COX-2 and iNOS protein expression was upregulated in the jejunum of mice in the model group when compared to the control group, and Chaishen Jieyu Decoction could downregulate iNOS protein expression in jejunum (P < 0.01), and the mouse jejunum mRNA expression of ZO-1 and OCCLUDIN in model group were significantly decreased when compared with the control group(P < 0.01), while CSJY Decoction could increase these two proteins' mRNA expression(P < 0.05, P << 0.01);Also, compared with the control group, the pathological alterations such as incomplete jejunal structural organization, disorganized intestinal epithelial cell arrangement, crypt damage as well as intestinal epithelial integrity in the model group mice tended to be normalized after administration of CSJY Decoction. the composition and content of fecal bile acids in IBS-D mice were altered, after Chaishen Jieyu Decoction intervention, the disturbed fecal bile acid metabolism and disposition were significantly altered, especially for lithic cholic acid (LCA), deoxycholic acid (DCA) and deoxycholic acid (CDCA). CONCLUSION Chaishen Jieyu Decoction has therapeutic effect on IBS-D mice and this effect may be partially contributed to regulating the bile acids metabolism.
- irritable bowel syndrome /
- Chaishen Jieyu Decoction /
- bile acid /
- intestinal mucosal barrier /
- metabolic profile

HTML全文

图 1 柴参解郁汤对IBS-D小鼠空肠病理损伤、粪便含水量及空肠绒毛长度的影响

注: 与模型组比较，^**P < 0.01。x±s, n=8。

Figure 1. The effects of Chaishen Jieyu Decoction on pathological changes of mouse jejunum segment, fecal water content and villi length of jejunum

下载: 全尺寸图片幻灯片

图 2 柴参解郁汤对IBS-D小鼠胃肠道通透性及紧密连接蛋白ZO-1、OCCLUDIN mRNA表达的影响

注：与模型组比较，^*P < 0.05，^**P < 0.01。x±s，n=4。

Figure 2. The effects of Chaishen Jieyu Decoction on the intestinal permeability and the relative mRNA expressions of main tight junctions proteins ZO-1, OCCLUDIN in mice

下载: 全尺寸图片幻灯片

图 3 各组小鼠空肠中COX-2、iNOS蛋白的表达情况

注：与模型组比较，^*P < 0.05, ^**P < 0.01。x±s，n=4。

Figure 3. The relative protein expressions of COX-2 and iNOS in each group

下载: 全尺寸图片幻灯片

图 4 各组小鼠粪便中胆汁酸的的OPLS-DA(A)和聚类热图(B)

Figure 4. OPLS-DA score scatter plot (A) and the heatmap (B) of fecal bile acid in different groups

下载: 全尺寸图片幻灯片

图 5 各组小鼠粪便中的差异胆汁酸的绝对定量结果

注: 与模型组比较，^*P < 0.05，^{* *}P < 0.01。x±s，n=8。

Figure 5. The absolute quantitative results of the changed fecal bile acids in different groups

下载: 全尺寸图片幻灯片

表 1 引物序列

Table 1. The sequences of the primers

引物名称	上游序列	下游序列
ZO-1	5'-ACTCCCACTTCCCCAAAAAC-3'	5'-CCACAGCTGAAGGACTCACA-3'
Occludin	5'-ACTGGGTCAGGGAATATCCA-3'	5'-TCAGCAGCAGCCATGTACTC-3'
GAPDH	5'-AGGTCGGTGTGAACGGATTTG-3'	5'-TGTAGACCATGTAGTTGAGGTCA-3'

下载: 导出CSV

参考文献(17)

[1]	RASKOV H, BURCHARTH J, POMMERGAARD HC, et al. Irritable bowel syndrome, the microbiota and the gut-brain axis[J]. Gut Microbes, 2016, 7(5): 365-383. doi: 10.1080/19490976.2016.1218585
[2]	PERINO A, DEMAGNY H, VELAZQUEZ-VILLEGAS L, et al. Molecular physiology of bile acid signaling in health, disease, and aging[J]. Physiol Rev, 2021, 101(2): 683-731. doi: 10.1152/physrev.00049.2019
[3]	HOLTMANN GJ, FORD AC, TALLEY NJ. Pathophysiology of irritable bowel syndrome[J]. Lancet Gastroenterol Hepatol, 2016, 1(2): 133-146. doi: 10.1016/S2468-1253(16)30023-1
[4]	WARD JBJ, LAJCZAK NK, KELLY OB, et al. Ursodeoxycholic acid and lithocholic acid exert anti-inflammatory actions in the colon[J]. Am J Physiol Gastrointest Liver Physiol, 2017, 312(6): G550-G558. doi: 10.1152/ajpgi.00256.2016
[5]	冯文林, 伍海涛. 从"魄门亦为五脏使"探讨肠易激综合征的发病[J]. 中国中医基础医学杂志, 2016, 22(10): 1314-1316. https://www.cnki.com.cn/Article/CJFDTOTAL-ZYJC201610014.htm FENG WL, WU HT. Exploring the pathogenesis of irritable bowel syndrome based on the theory of "anus being envoy of five zang-organs"[J]. Chin J Basic Med Tradit Chin Med, 2016, 22(10): 1314-1316. https://www.cnki.com.cn/Article/CJFDTOTAL-ZYJC201610014.htm
[6]	郭妍. 柴参解郁汤加减治疗腹泻型肠易激综合征肝郁脾虚证的临床疗效观察[D]. 南京: 南京中医药大学, 2020. GUO Y. The clinical observation of Chaishen Jieyu Decoction on treating the diarrhea type of irritable bowel syndrome with liver depression and spleen deficiency[D]. Nanjing: Nanjing University of Chinese Medicine, 2020.
[7]	赵迎盼, 唐旭东, 卞兆祥, 等. IBS-D肝郁脾虚型病证症结合大鼠模型的建立与评价的初步研究[J]. 中国中西医结合杂志, 2013, 33(11): 1507-1514. doi: 10.7661/CJIM.2013.11.1507 ZHAO YP, TANG XD, BIAN ZX, et al. Preliminary study of establishing and assessing IBS-D model rats of Gan stagnation and pi deficiency syndrome[J]. Chin J Integr Tradit West Med, 2013, 33(11): 1507-1514. doi: 10.7661/CJIM.2013.11.1507
[8]	KUS K, KIJ A, ZAKRZEWSKA A, et al. Alterations in arginine and energy metabolism, structural and signalling lipids in metastatic breast cancer in mice detected in plasma by targeted metabolomics and lipidomics[J]. Breast Cancer Res, 2018, 20: 148. doi: 10.1186/s13058-018-1075-y
[9]	MOTTACKI N, SIMREN M, BAJOR A. Review article: Bile acid diarrhoea-pathogenesis, diagnosis and management[J]. Aliment Pharmacol Ther, 2016, 43(8): 884-898. doi: 10.1111/apt.13570
[10]	TICHO AL, POOJA M, DUDEJA PK, et al. Intestinal absorption of bile acids in health and disease[J]. Compr Physiol, 2019, 10(1): 21-56.
[11]	JEFFERY IB, DAS A, O'HERLIHY E, et al. Differences in fecal microbiomes and metabolomes of people with vs without irritable bowel syndrome and bile acid malabsorption[J]. Gastroenterology, 2020, 158(4): 1016-1028.e8. doi: 10.1053/j.gastro.2019.11.301
[12]	居永慧, 姚卫峰, 张丽. 胆汁酸代谢在中药研究中的应用进展[J]. 中国中药杂志, 2020, 45(10): 2360-2367. https://www.cnki.com.cn/Article/CJFDTOTAL-ZGZY202010020.htm JU YH, YAO WF, ZHANG L. Progress in application of bile acid metabolism in traditional Chinese medicine study[J]. China J Chin Mater Med, 2020, 45(10): 2360-2367. https://www.cnki.com.cn/Article/CJFDTOTAL-ZGZY202010020.htm
[13]	SINHA SR, HAILESELASSIE Y, NGUYEN LP, et al. Dysbiosis-induced secondary bile acid deficiency promotes intestinal inflammation[J]. Cell Host Microbe, 2020, 27(4): 659-670. doi: 10.1016/j.chom.2020.01.021
[14]	VIJAYVARGIYA P, CAMILLERI M, BURTON D, et al. Bile and fat excretion are biomarkers of clinically significant diarrhoea and constipation in irritable bowel syndrome[J]. Aliment Pharmacol Ther, 2019, 49(6): 744-758. doi: 10.1111/apt.15106
[15]	VIJAYVARGIYA P, CAMILLERI M, CARLSON P, et al. Performance characteristics of serum C4 and FGF19 measurements to exclude the diagnosis of bile acid diarrhoea in IBS-diarrhoea and functional diarrhoea[J]. Aliment Pharmacol Ther, 2017, 46(6): 581-588. doi: 10.1111/apt.14214
[16]	ZHAO L, YANG W, CHEN Y, et al. A Clostridia-rich microbiota enhances bile acid excretion in diarrhea-predominant irritable bowel syndrome[J]. J Clin Investig, 2019, 130(1): 438-450. doi: 10.1172/JCI130976
[17]	FIORUCCI S, DISTRUTTI E. Bile acid-activated receptors, intestinal microbiota, and the treatment of metabolic disorders[J]. Trends Mol Med, 2015, 21(11): 702-714.