Effect of Tongxie Yaofang on IL-6, IL-6R mRNA and Protein Expressions of Hypothalamus Tissues in Rats with Experimental Ulcerative Colitis

LI Ting; ZHU Xiang-dong; YANG Yi; ZHAI Yan-hui

doi:10.13422/j.cnki.syfjx.2017110103

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Effect of Tongxie Yaofang on IL-6, IL-6R mRNA and Protein Expressions of Hypothalamus Tissues in Rats with Experimental Ulcerative Colitis

更新时间：2024-01-04

- Effect of Tongxie Yaofang on IL-6, IL-6R mRNA and Protein Expressions of Hypothalamus Tissues in Rats with Experimental Ulcerative Colitis
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 23, Issue 11, Pages: 103-108(2017)
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- DOI：10.13422/j.cnki.syfjx.2017110103
  CLC： R285.5
- Published：2017
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LI Ting, ZHU Xiang-dong, YANG Yi, et al. Effect of Tongxie Yaofang on IL-6, IL-6R mRNA and Protein Expressions of Hypothalamus Tissues in Rats with Experimental Ulcerative Colitis[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(11): 103-108.
DOI：

LI Ting, ZHU Xiang-dong, YANG Yi, et al. Effect of Tongxie Yaofang on IL-6, IL-6R mRNA and Protein Expressions of Hypothalamus Tissues in Rats with Experimental Ulcerative Colitis[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(11): 103-108. DOI： 10.13422/j.cnki.syfjx.2017110103.

摘要

目的：观察痛泻要方对溃疡性结肠炎（ulcerative colitis，UC）大鼠下丘脑中白细胞介素-6（IL-6）和白细胞介素-6受体（IL-6R）基因和蛋白表达的影响。方法：将SPF级Wistar大鼠运用复合方法进行溃疡性结肠炎造模后，随机分为 5 组：模型组，痛泻要方低、中、高剂量（2.75，5.5，11 g·kg-1）组，美沙拉嗪（5-ASA）组。药物处理大鼠21 d后麻醉处死，肉眼观察结肠大体形态损伤并进行评分。分离各组大鼠血清，酶联免疫吸附试验（enzyme linked immunosorbent assay，ELISA）检测血清中IL-6含量，分离大鼠下丘脑组织并提取总RNA后，实时荧光定量PCR（Real-time polymerase chain reaction，Real-time PCR）法检测 IL-6和IL-6R基因表达，免疫组织化学染色法检测下丘脑组织IL-6和IL-6R的分布及蛋白表达量。结果：ELISA结果显示，与空白组比较，模型组大鼠血清中IL-6含量明显升高，经过药物处理后，药物组血清中IL-6含量均有所下降，其中以痛泻要方高剂量组效果最为明显（P<0.05）。Real-time PCR结果表明，模型组大鼠下丘脑组织中IL-6和IL-6R基因表达量明显高于空白组（P<0.05），经药物治疗后，药物组IL-6和IL-6R基因表达均有所降低，其中以5-ASA组和痛泻要方高剂量组降低最为明显（P<0.05）。免疫组织化学染色结果表明，与空白组比较，溃疡性结肠炎大鼠IL-6和IL-6R蛋白表达量明显增加（P<0.05），药物处理可以降低大鼠下丘脑IL-6和IL-6R蛋白表达量，其中以5-ASA组和痛泻要方高剂量组降低最为明显（P<0.05）。结论：痛泻要方对 2，4，6三硝基苯磺酸（2，4，6-trinitrobenzene sulfonic acid，TNBS）/乙醇法UC大鼠下丘脑组织中IL-6和IL-6R基因和蛋白的表达具有明显抑制作用，提示痛泻要方治疗溃疡性结肠炎的机制可能与抑制下丘脑中IL-6/IL-6R信号通路的异常活化有关。

Abstract

Objective: To explore the curative effect of Tongxie Yaofang on ulcerative colitis (UC) in rat hypothalamus cytokine interleukin-6 (IL-6) and interleukin-6 receptors (IL-6R) gene and protein expressions. Method: After the ulcerative colitis model was established by the composite method. SPF Wistar rats were randomly divided into 5 groups: model group

low

middle

high-dose Tongxie Yaofang groups (2.75

5.5

11 g·kg-1)

and 5-ASA group. After modeling

rats were given drugs for 21 days. Then

colonic mucosa damage was observed. The levels of IL-6 in the serum tissues were detected by enzyme linked immunosorbent assay (ELISA). The proteins expressions of IL-6 and IL-6R in rat hypothalamus tissues were detected by immunohistochemistry

and their gene expressions were measured by Real-time PCR method. Result: ELISA test results show that compared with control group

model group showed significant increase in content of IL-6 in serum of rats. After drug treatment

the serum IL-6 in drug group were decreased

with the most obvious effect in high-dose Tongxie Yaofang group (P<0.05). Real-time PCR results showed that model group's hypothalamic IL-6 and IL-6R gene expressions were higher than those of blank group (P<0.05). After treatment with drugs

drug group's IL-6 and IL-6R gene expressions were reduced

and 5-ASA and high-dose Tongxie Yaofang groups were superior to the other groups (P<0.05). Immunohistochemical staining results showed that compared with control group

IL-6 and IL-6R protein expressions of ulcerative colitis rats increased significantly (P<0.05)

drug treatment can reduce their hypothalamic IL-6 and IL-6R protein expressions

and 5-ASA and high-dose Tongxie Yaofang groups were superior to the other groups (P<0.05). Conclusion: Tongxie Yaofang can obviously inhibit IL-6 and IL-6R gene and protein expressions in hypothalamic tissues of the rat UC model by TNBS/ethanol method

suggesting Tongxie Yaofang's mechanism in the treatment of ulcerative colitis may be correlated with inhibition of abnormal activation of IL-6/IL-6R signaling pathway in hypothalamus.

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