Effect of Warming and Heat-clearing Method on Expressions of FoxP3 and ROR-<italic style="font-style: italic">γ</italic>t in Colon Tissue of Rats with Intestinal Endotoxemia Caused by Acute-on-chronic Liver Failure

Bin CHEN; Xiu-li DING; Jie PENG; Yun-fang LI; Ruo-yu WANG; Yi ZHOU

doi:10.13422/j.cnki.syfjx.20200225

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Effect of Warming and Heat-clearing Method on Expressions of FoxP3 and ROR-γt in Colon Tissue of Rats with Intestinal Endotoxemia Caused by Acute-on-chronic Liver Failure

Research on Immunological Mechanism of TCM Anti-inflammatory Response to Liver Failure | 更新时间：2021-04-16

- Effect of Warming and Heat-clearing Method on Expressions of FoxP3 and ROR-γt in Colon Tissue of Rats with Intestinal Endotoxemia Caused by Acute-on-chronic Liver Failure
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 26, Issue 2, Pages: 19-25(2020)
- 作者机构：
  
  1.湖南中医药大学　第一附属医院，长沙　 410007
  2.湖南中医药大学，长沙　 410208
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20200225
  CLC： R22; R242; R2-031;R285.5
- Received：21 June 2019，
  
  Published Online：11 October 2019，
  
  Published：20 January 2020
- 稿件说明：
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Bin CHEN, Xiu-li DING, Jie PENG, et al. Effect of Warming and Heat-clearing Method on Expressions of FoxP3 and ROR-γt in Colon Tissue of Rats with Intestinal Endotoxemia Caused by Acute-on-chronic Liver Failure[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(2): 19-25.
DOI：

Bin CHEN, Xiu-li DING, Jie PENG, et al. Effect of Warming and Heat-clearing Method on Expressions of FoxP3 and ROR-γt in Colon Tissue of Rats with Intestinal Endotoxemia Caused by Acute-on-chronic Liver Failure[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(2): 19-25. DOI： 10.13422/j.cnki.syfjx.20200225.

摘要

目的：

研究清温并用法(即温阳解毒化瘀方)对慢加急性肝衰竭(ACLF)大鼠结肠组织叉头样转录因子3(FoxP3)，维甲酸相关核孤儿受体-

t(ROR-

t)表达的影响，探索其对肝衰竭肠源性内毒素血症(IETM)的可能调节机制。

方法：

130只SD大鼠随机分为正常组10只，造模组120只。造模组以皮下注射牛血清白蛋白致敏法，腹腔注射

-氨基半乳糖(

-Gal)＋脂多糖(LPS)建立ACLF大鼠模型。将成模的大鼠随机分为模型组，清法组(茵陈蒿汤6.68 g·kg

-1

)，温法组(茵陈术附汤7.09 g·kg

-1

)，清温并用法组(温阳解毒化瘀方19.53 g·kg

-1

)。正常组及模型组予蒸馏水灌胃，其余各组予等容积相应中药灌胃，共1周。检测各项指标在1，12，24 h时间点的数值，流式细胞仪检测外周血调节性T淋巴细胞(Treg)/辅助性T淋巴细胞(Th)17细胞比值变化，实时荧光定量聚合酶链式反应(Real-time PCR)检测结肠组织FoxP3，ROR-

t mRNA表达，原位杂交法、免疫组化法观察大鼠结肠组织FoxP3，ROR-

t mRNA及蛋白表达。

结果：

与正常组比较，模型组大鼠Treg/Th17细胞在上述时间点显著下降(

0.01)；与模型组比较，清法、温法组Treg/Th17细胞差异无统计学意义，清温并用法Treg/Th17细胞升高(

0.05)；与正常组比较，模型组大鼠FoxP3，ROR-

t mRNA在上述时间点表达均显著升高(

0.01)；与模型组比较，清法、温法组FoxP3，ROR-

t mRNA在上述时间点均有所降低(

0.05)，清温并用法组显著降低(

0.01)；清温并用法组FoxP3，ROR-

t mRNA较清法组、温法组有所下降(

0.05)。

结论：

清温并用法治疗肝衰竭IETM的机制与可能与调节FoxP3，ROR-

t表达有关。

Abstract

Objective:

To study the effect of warming and heat-clearing method (Wenyang Jiedu Huayu decoction) on the expressions of Forkhead box P3 (FoxP3)

Retinoic acid-related orphan receptor gamma t (ROR-

t) in colon tissue of mice with acute-on-chronic liver failure (ACLF)

in order to explore the possible regulatory mechanism on intestinal endotoxemia (IETM) in liver failure mice.

Method:

The 130 SD rats were randomly divided into normal group (10 rats) and model group (120 rats). The ACLF mice model was established through the subcutaneous injection with bovine serum albumin and the intraperitoneal injection with

-galactosamine(

-Gal) and lipopolysaccharide (LPS). The model mice were randomly divided into model group

heat-clearing group (Yinchenhao decoction

6.68 g·kg

-1

)

warming group (Yinchen Zhufu decoction

7.09 g·kg

-1

) and warming and heat-clearing group (Wenyang Jiedu Huayu decoction

19.53 g·kg

-1

). The normal group and the model group were given distilled water by gastric lavage

while the other groups were given equal volume of corresponding Chinese herbal medicines for a week. The value of each index at 1

12 and 24 h was measured. The ratio of Treg/Th17 cell in peripheral blood were detected and calculated by flow cytometry. Real-time fluorescence quantitative PCR (Real-time PCR) was used to detect the expressions of FoxP3 and ROR-

t in colon tissues of mice at different time points. In situ hybridization and immunohistochemistry were used to observe the expressions of FoxP3 and ROR-

t genes and proteins.

Result:

Compared with normal group

the ratio of Treg/Th17 in the model group decreased significantly at each time point (

0.01). Compared with the model group

the Treg/Th17 ratio increased only in the warming and heat-clearing method group (

0.05). Compared with normal group

the expression of ROR-

t in the model group was significantly higher (

0.01)

and the expression of ROR-

t in the model group was higher than FoxP3.Compared with the model group

the expressions of FoxP3 and ROR-

t mRNA in the heat-clearing group and the warming group decreased at each time point (

0.05)

and the expressions of FoxP3 and ROR-

t in the warming and heat-clearing method group decreased significantly (

0.01). The expressions of FoxP3 and ROR-

t mRNA in warming and heat-clearing group decreased compared with those in the warming group and heat-clearing group (

0.05).

Conclusion:

The mechanism of the warming and heat-clearing method on IETM in liver failure may be related to the regulation of FoxP3 and ROR-

t expressions.

关键词

Keywords

references

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Effect of Warming and Heat-clearing Method on Expressions of FoxP3 and ROR-γt in Colon Tissue of Rats with Intestinal Endotoxemia Caused by Acute-on-chronic Liver Failure

DOI：10.13422/j.cnki.syfjx.20200225

摘要

Abstract

关键词

Keywords

references