绿原酸通过TLR4/NF-κB信号通路调控脓毒症大鼠肠上皮细胞屏障功能的研究

王金鑫; 饶春晖; 代波; 俞慧琴; 汪志刚

doi:10.16766/j.cnki.issn.1674-4152.002894

绿原酸通过TLR4/NF-κB信号通路调控脓毒症大鼠肠上皮细胞屏障功能的研究

doi: 10.16766/j.cnki.issn.1674-4152.002894

1.
浙江省医疗健康集团杭州医院消化内科，浙江杭州 310022
2.
杭州市中医院肛肠科，浙江杭州 310000

基金项目:

浙江省医药卫生科技计划项目 2019KY522

详细信息

通讯作者:
王金鑫, E-mail: 2781174394@qq.com

中图分类号: R631R-332
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- 被引次数: 0
出版历程
- 收稿日期: 2022-04-20
- 网络出版日期: 2023-04-19

Chlorogenic acid effects of intestinal epithelial barrier function with septic rats through TLR4/NF-κB signaling pathway

1.
Department of Gastroenterology, Zhejiang Medical Health Group Hangzhou Hospital, Hangzhou, Zhejiang 310022, China

摘要

摘要: 目的观察绿原酸通过调控TLR4/NF-κB信号通路减轻脓毒症(sepsis)大鼠肠道氧化应激损伤、改善肠上皮细胞屏障功能的作用。方法选取SD大鼠60只，采用随机数字表法分成假手术组、模型对照组、绿原酸(低、高剂量)组，每组15只。术后24 h，应用酶联免疫吸附法(ELISA)检测大鼠血清中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、IL-10；比色法测定小肠组织中一氧化氮(NO)、超氧化物歧化酶(SOD)、丙二醛(MDA)水平；采用Western blotting法测定小肠组织中的蛋白表达[密封蛋白-1(Claudin-1)、闭锁连接蛋白-1(ZO-1)及TLR-4/NF-κB通路相关的蛋白]。结果与模型对照组[(4.45±0.14)分]比较，低剂量组[(4.17±0.22)分]、高剂量组[(1.95±0.08)分]Chiu评分明显下降(t=4.159、60.048，均P < 0.001)；高剂量组血清TNF-α、IL-6、IL-10水平及小肠组织中NO、MDA含量均显著降低(均P < 0.05)，SOD含量、Claudin-1、ZO-1蛋白表达量及MyD88、TLR-4蛋白与p-NF-κB p65(Ser536)水平均显著升高(均P < 0.05)。结论绿原酸可经抑制TLR4/NF-kB信号转导途径，减轻脓毒症大鼠的炎症反应程度及小肠组织的氧化应激损伤，进而促进大鼠的肠道屏障功能改善。
- 绿原酸 /
- TLR4/NF-κB信号通路 /
- 肠道菌群 /
- 肠上皮细胞屏障功能
Abstract: Objective To observe the effect of chlorogenic acid on reducing intestinal oxidative stress injury and improving intestinal cell barrier function through TLR4/NF-κB signaling pathway in septic rats. Methods Sixty SD rats were randomly divided into four groups by the random number table method: sham operation group, model control group and chlorogenic acid (low and high dose) group, with 15 rats in each group. 24 hours after operation, the levels of tumor necrosis factor-α (TNF-α), interleukin (IL-6) and IL-10 were detected by enzyme-linked immunosorbent assay (ELISA), the levels of nitric oxide (NO), superoxide dismutase (SOD) and malondialdehyde (MDA) in small intestine were measured by colorimetry, and western blot was used to detect claudin-1, ZO-1 and TLR-4/NF-κB pathway related protein expression. Results Compared with the model control group, Chiu score in low dose group (4.17±0.22) and high dose group (1.95±0.08) decreased significantly (t=4.159, 60.048, all P < 0.01). The levels of TNF-α, IL-6, IL-10 and the contents of NO and MDA in the small intestine in high dose group were significantly decreased (all P < 0.05), the content of SOD, the expression of Claudin-1, ZO-1 protein, MyD88, TLR-4 protein and p-NF-κB p65 (Ser536) increased significantly (all P < 0.05). Conclusion Chlorogenic acid can reduce serum inflammatory response, reduce oxidative damage of small intestine in septic rats through inhibiting TLR4/NF-κB signal transduction, and then improve intestinal barrier function.
- Chlorogenic acid /
- TLR4/NF-κB signal pathway /
- Intestinal flora /
- Intestinal epithelial barrier function

HTML全文

图 1 绿原酸对小肠组织中Claudin-1和ZO-1表达的影响

Figure 1. Effect of chlorogenic acid on Claudin-1 and ZO-1 expression in small intestinal tissue

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图 2 绿原酸对脓毒症大鼠TLR-4/NF-κB信号通路的影响

Figure 2. Effect of chlorogenic acid on TLR-4/NF-κB signaling pathway in septic rats

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表 1 4组脓毒症大鼠血清TNF-α、IL-6和IL-10水平比较(x±s，pg/mL)

Table 1. Comparison of serum TNF-α, IL-6, and IL-10 levels in four septic rats(x±s, pg/mL)

组别	只数	TNF-α	IL-6	IL-10
假手术组	15	41.38±9.02	76.09±20.36	7.04±2.03
模型对照组	15	106.48±14.25	249.61±31.58	20.16±5.84
低剂量组	15	97.47±15.18	237.68±28.39	18.72±4.25
高剂量组	15	74.29±10.80	202.73±25.44	15.66±3.19
F值		80.142	132.320	31.185
P值		＜0.001	＜0.001	＜0.001

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表 2 4组脓毒症大鼠小肠组织NO、SOD和MDA水平比较(x±s)

Table 2. Comparison of the levels of NO, SOD, and MDA in small intestinal tissue of septic rats among the four groups(x±s)

组别	只数	NO(μmol/L)	SOD(U/mL)	MDA(nmol/mL)
假手术组	15	39.65±7.38	43.97±9.81	98.37±11.86
模型对照组	15	110.22±13.91	19.62±5.74	210.08±17.63
低剂量组	15	77.36±10.13	23.65±7.19	198.71±20.39
高剂量组	15	65.36±8.87	38.93±9.06	123.08±13.15
F值		119.970	31.446	175.821
P值		＜0.001	＜0.001	＜0.001

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