Explore Effect of Jieyu Qutan Huazhuo Prescription on Gut-liver Axis of Rats with High-fat Diet Based on 16S rDNA Sequencing

Na LI; Yang-yang WU; Jin-long DUAN; Xu-tong ZHENG; Kui-wu YAO

doi:10.13422/j.cnki.syfjx.20210327

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Explore Effect of Jieyu Qutan Huazhuo Prescription on Gut-liver Axis of Rats with High-fat Diet Based on 16S rDNA Sequencing

Pharmacology | 更新时间：2021-04-02

- Explore Effect of Jieyu Qutan Huazhuo Prescription on Gut-liver Axis of Rats with High-fat Diet Based on 16S rDNA Sequencing
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 27, Issue 9, Pages: 77-85(2021)
- 作者机构：
  
  1.中国中医科学院广安门医院，北京 100053
  2.洛阳市中医院，河南洛阳 471000
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20210327
  CLC： R22;R242;R2-031;R285.5
- Received：15 November 2020，
  
  Published Online：15 December 2020，
  
  Published：05 May 2021
- 稿件说明：
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李娜,吴阳阳,段锦龙等.基于16S rDNA测序的肠道菌群探讨解郁祛痰化浊方对高脂饮食大鼠肠-肝轴的影响[J].中国实验方剂学杂志,2021,27(09):77-85.

LI Na,WU Yang-yang,DUAN Jin-long,et al.Explore Effect of Jieyu Qutan Huazhuo Prescription on Gut-liver Axis of Rats with High-fat Diet Based on 16S rDNA Sequencing[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(09):77-85.
李娜,吴阳阳,段锦龙等.基于16S rDNA测序的肠道菌群探讨解郁祛痰化浊方对高脂饮食大鼠肠-肝轴的影响[J].中国实验方剂学杂志,2021,27(09):77-85. DOI： 10.13422/j.cnki.syfjx.20210327.

LI Na,WU Yang-yang,DUAN Jin-long,et al.Explore Effect of Jieyu Qutan Huazhuo Prescription on Gut-liver Axis of Rats with High-fat Diet Based on 16S rDNA Sequencing[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(09):77-85. DOI： 10.13422/j.cnki.syfjx.20210327.

摘要

目的

研究解郁祛痰化浊方（JQHP）对高脂饮食大鼠肠道菌群的影响，探讨中药调控肠道微生物群进而恢复肠-肝轴的平衡。

方法

将70只雄性SPF级别Wistar大鼠随机分为正常组（10只）与模型组（60只），正常组饲喂正常饲料，模型组饲喂高脂饲料。12周后将模型组随机分为6组，每组10只，即模型组，血脂康组，立普妥组，JQHP低、中、高剂量组。JQHP低、中、高剂量组分别灌胃JQHP颗粒剂0.4，0.8，1.6 g·kg

-1

，立普妥组予立普妥2 mg·kg

-1

，血脂康组予血脂康0.1 g·kg

-1

，正常组和模型组大鼠灌胃同等量蒸馏水，连续灌胃8周后收集粪便，进行16S rRNA基因测序，行腹主动脉取血检测血脂，取肝脏组织及回肠组织苏木素-伊红（HE）染色后进行病理形态学观察。

结果

与正常组比较，模型组大鼠血脂四项总胆固醇（TC），甘油三酯（TG），低密度脂蛋白胆固醇（LDL-C）均见明显升高，高密度脂蛋白胆固醇（HDL-C）降低（

0.01）；与模型组比较，血脂康组、立普妥组TC，TG显著性下降（

0.01），血脂康组HDL-C升高（

0.05）。JQHP中剂量组较模型组对肝脏脂肪样变有一定的缓解作用，可以减轻炎性细胞的浸润情况。JQHP可使回肠结构淋巴组织增生情况好转，且中剂量组疗效最显著。Shannon曲线结果表明，与正常组比较，JQHP中剂量组显著提高（

0.01）；与模型组比较，JQHP中、高剂量组明显升高（

0.05，

0.01）；与JQHP中剂量组比较，其他用药组降低（

0.05，

0.01）。主成分多样性分析（PCA）示中药中剂量组多样性和丰度高于其他用药组。线性判别分析（LDA）中，与正常组比较，模型组拟杆菌纲，瘤胃球菌科，拟杆菌S24-7，瘤胃球菌UCG-005下调（

0.01），脱硫弧菌目、丹毒丝菌目、毛螺菌科上调（

0.05，

0.01）。与模型组比较，JQHP中剂量组的拟杆菌纲，瘤胃球菌科，拟杆菌S24-7，瘤胃球菌UCG-005上调（

0.05，

0.01），丹毒丝菌目下调（

0.01）。与JQHP中剂量组比较，其他用药组拟杆菌纲，瘤胃球菌科，拟杆菌S24-7，瘤胃球菌UCG-005中降低（

0.05，

0.01），在丹毒丝菌目、毛螺菌科中升高（

0.05，

0.01）。

结论

JQHP调整肠道物种丰度和多样性，改善肝脏组织和回肠黏膜状态，调节血脂水平，恢复正常肠道生态环境，可能与调节与炎症相关的肠道菌群而恢复肠-肝轴平衡有关，以中剂量组效果最佳。

Abstract

Objective

To study the effect of Jieyu Qutan Huazhuo prescription（JQHP） on the gut microbiota of rats with high-fat diet，and to explore the effect of Chinese medicine on the regulation of gut microflora and the restoration of gut-liver axis balance.

Method

Seventy male SPF Wistar rats were randomly divided into a normal group of 10 and a model group of 60. Mice in the normal group were fed with normal diet and mice in the model group were fed high-fat diet. After 12 weeks，the model group was randomly divided into 6 groups with 10 animals in each group，namely the model group，Xuezhikang group，Liputuo group，and low，medium and high-dose groups of JQHP. The JQHP low-dose，medium-dose and high-dose rats were intragastrically daministered with 0.4，0.8，1.6 g·kg

-1

，respectively， rats in Liputuo group with Liputuo 2 mg·kg

-1

，rats in Xuezhikang group with Xuezhikang 0.1 g·kg

-1

. The rats in the normal group and the model group were intragastrically administered with the same amount of distilled water. Stool were collected after continuous gavaging for 8 weeks，16S rRNA gene sequencing was performed，blood was collected from the abdominal aorta to detect blood lipids，and the liver tissue and ileum tissue were collected for hematoxylin and eosin （HE） staining for pathomorphological observation.

Result

Compared with the normal group，the total cholesterol （TC），triglyceride （TG），and low-density lipoprotein （LDL-C） in the model group were significantly increased，while the high-density lipoprotein （HDL-C） was decreased （

0.01）. Compared with the model group，TC and TG values were decreased significantly in Xuezhikang group （

0.01），HDL-C value was increased （

0.05），and in the Liputuo group TC and TG were decreased significantly （

0.01）. Compared with the model group，the middle-dose group of JQHP had a certain alleviating effect on liver steatosis and could reduce the infiltration of inflammatory cells. The JQHP could improve the proliferation of lymphoid tissues in the ileal structure，and the middle-dose group has the most significant effect. The results of Shannon curve showed that compared with the normal group，the middle-dose group of JQHP increased significantly （

0.01）. Compared with the model group，the middle and high-dose group of JQHP increased （

0.05，

0.01）. Compared with the middle-dose group of JQHP，the other drug group decreased （

0.05，

0.01）. Principal component diversity analysis（PCA） showed that the diversity and abundance in the middle-dose JQHP group were higher than those in other drug groups. In linear discriminant analysis（LDA），compared with the normal group，Bacteroidia，Ruminococcaceae，

Bacteroides

S24-7，and

Rumenococcus

UCG-005 were down-regulated in the model group（

0.01），while the orders of Desulfovibrionales，Erysipelotrichales and

Lachnospiraceae were up-regulated in the model group （

0.05，

0.01）. Compared with the model group，the Bacteroidia，Ruminococcaceae，

Bacteroides

S24-7，and

Rumencoccus

UCG-005 in the middle-dose JQHP group were increased （

0.05，

0.01），and the orders of Erysipelotrichales were decreased （

0.01）. Compared with the middle-dose JQHP group，Bacteroidia，Ruminococcaceae，

Bacteroides

S24-7，and

Rumencoccus

UCG-005 in other drug groups were reduced（

0.05，

0.01），and the order of Erysipelotrichales and Lachnospiraceae were increased （

0.05，

0.01）.

Conclusion

JQHP can regulate the abundance and diversity of the gut microbiota，improve the state of liver tissue and ileum mucosa，regulate blood lipid levels，and restore the normal intestinal ecological environment. It may be related to the regulation of inflammation-related gut microbiota in order to restore the balance of the gut-liver axis，and the middle-dose JQHP group has the best effect.

关键词

Keywords

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