Effect of Qingchang Huashi Granule on AR/-arrestin2/NF-B Signaling Pathway in Ulcerative Colitis

DAI Lu-ming; ZHU Lei; SHEN Hong

doi:10.13422/j.cnki.syfjx.20180837

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Effect of Qingchang Huashi Granule on AR/-arrestin2/NF-B Signaling Pathway in Ulcerative Colitis

更新时间：2024-01-04

- Effect of Qingchang Huashi Granule on AR/-arrestin2/NF-B Signaling Pathway in Ulcerative Colitis
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 9, Pages: 86-94(2018)
- 作者机构：
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20180837
  CLC： R285.5
- Published：2018
- 稿件说明：
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DAI Lu-ming, ZHU Lei, SHEN Hong. Effect of Qingchang Huashi Granule on AR/-arrestin2/NF-B Signaling Pathway in Ulcerative Colitis[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(9): 86-94.
DOI：

DAI Lu-ming, ZHU Lei, SHEN Hong. Effect of Qingchang Huashi Granule on AR/-arrestin2/NF-B Signaling Pathway in Ulcerative Colitis[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(9): 86-94. DOI： 10.13422/j.cnki.syfjx.20180837.

摘要

目的：观察清肠化湿颗粒对溃疡性结肠炎（ulcerative colitis，UC）模型大鼠及人结肠癌上皮细胞株HT-29炎症模型β2肾上腺素受体（β2AR）/β-抑制蛋白2（β-arrestin2）/核转录因子-κB（NF-κB）信号通路的干预作用。方法：取8只大鼠作为空白组，其余大鼠采用三硝基苯磺酸（trinitro-benzene-sulfonic acid，TNBS）灌肠法复制UC大鼠模型；待模型建立成功后，随机分为模型组，柳氮磺胺吡啶（SASP，1.0 g ·kg-1）组，清肠化湿颗粒低、中、高剂量（2.8，5.5，11.0 g ·kg-1）组，每组8只，灌胃给药10 d，每日1次；酶联免疫吸附法（enzyme linked immunosorbent assay，ELISA）测定UC大鼠结肠组织中巨噬细胞移动抑制因子（MIF），基质金属蛋白酶-1（MMP-1），MMP-2，MMP-9，胰岛素样生长因子-1（IGF-1），超氧化物歧化酶（SOD），丙二醛（MDA），髓过氧化物酶（MPO），一氧化氮（NO），一氧化氮合成酶（iNOS），谷胱甘肽-过氧化物酶（GSH-Px）；免疫组化法检测模型大鼠结肠黏膜β2AR，β-arrestin2，NF-κB定位表达；采用肿瘤坏死因子与脂多糖诱导HT-29细胞炎症模型；噻唑蓝（MTT）比色法检测清肠化湿颗粒对细胞增殖的影响，蛋白免疫印迹法（Western blot）检测细胞中β2AR，β-arrestin2，NF-κB蛋白表达量。结果：造模后，模型组大鼠较空白组大鼠结肠黏膜的结肠损伤分数明显升高；与模型组比较，3个剂量的清肠化湿颗粒给药组结肠黏膜损伤明显下降（P<0.05，P<0.01），大鼠结肠组织IGF-1，SOD，GSH-Px含量与β2AR，β-arrestin2表达明显上升，MIF，MMP-2，MMP-9，MDA，MPO，NO，iNOS含量及NF-κB表达明显下降（P<0.05，P<0.01）。结论：清肠化湿颗粒可缓解UC氧化应激反应及肠道炎症，其作用与激活β2AR/β-arrestin2/NF-κB信号通路有关。

Abstract

Objective: To study the effect of Qingchang Huashi granule on β2 adrenergic receptor(β2AR)/β-arrestin2/nuclear transcription factor-κB(NF-κB) signaling pathway in rat and human ulcerative colitis. Method: Eight rats were randomly selected as control group

while the other rats were used to duplicate the ulcerative colitis model induced by trinitrobenzene sulfonic acid; after successful modeling

the rats were randomly divided into model group

salazosulfapyridine(SASP) group (1.0 g · kg-1)

and low

medium and high-dose Qingchang Huashi granule groups (2.8

5.5

11.0 g · kg-1)

with 8 rats in each group. The drugs were given once a day by gavage for 15 days. The levels of macrophage mobile inhibitory factor(MIF)

matrix metalloproteinase-1(MMP-1)

macrophage migration inhibitory factor (MIF)

matrix metalloproteinase-1 (MMP-1)

MMP-2

MMP-9

insulin-like growth factor-1 (IGF-1)

superoxide dismutase (SOD)

malondialdehyde (MDA)

myeloperoxidase (MPO)

nitric oxide (NO)

nitric oxide synthase (iNOS)

glutathione peroxidase (GSH-Px) in experimental UC rats were measured by enzyme linked immunosorbent assay method(ELISA); immunohistochemical method was used to detect the location expressions of β2AR

β-arrestin2

NF-κB in rat models; tumor necrosis factor alpha and lipopolysaccharide were used to induce the HT-29 cell inflammation model; the effect of Qingchang Huashi granule on cell growth was detected by MTT method

the expressions of β2AR

β-arrestin2

NF-κB were measured with Western blot. Result: The injury score of colonic mucosa in model group rats was significantly higher than that of the normal group

expressions of IGF-1

SOD

GSH-Px

β2AR and β-arrestin2 decreased significantly

while expressions of MIF

MMP-2

MMP-9

MDA

MPO

iNOS and NF-κB increased in UC model group (P<0.05

P<0.01). After intervened with Qingchang Huashi granule

these indicators all recovered to normal. Conclusion: Qingchang Huashi granule can reduce the pathological process of ulcerative colitis through β2AR/β-arrestin2/NF-κB signaling pathway.

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