[1]马慧敏,杨丽红,金瑞林,等.慢性盆腔炎模型大鼠中miR-29 及炎症信号通路分子的表达水平及其作用机制研究[J].现代检验医学杂志,2022,37(06):14-18+109.[doi:10.3969/j.issn.1671-7414.2022.06.003]
　MA Hui-min,YANG Li-hong,JIN Rui-lin,et al.Study on the Expression Level and Mechanism of miR-29 and Inflammatory Signal Pathway Molecules in Chronic Pelvic Inflammatory Disease Model Rats[J].Journal of Modern Laboratory Medicine,2022,37(06):14-18+109.[doi:10.3969/j.issn.1671-7414.2022.06.003]
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慢性盆腔炎模型大鼠中miR-29 及炎症信号通路分子的表达水平及其作用机制研究()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第37卷
期数:
2022年06期
页码:
14-18+109
栏目:
论著
出版日期:
2022-11-15

文章信息/Info

Title:
Study on the Expression Level and Mechanism of miR-29 and Inflammatory Signal Pathway Molecules in Chronic Pelvic Inflammatory Disease Model Rats
文章编号:
1671-7414(2022)06-014-06
作者:
马慧敏; 杨丽红; 金瑞林; 王娟娟
(解放军陆军第81 集团军医院妇产科，河北张家口 075000)
Author(s):
MA Hui-min; YANG Li-hong; JIN Rui-lin; WANG Juan-juan
(Department of Obstetrics and Gynecology, the Hospital of the 81st Group Army of the PLA ，Hebei Zhangjiakou 075000，China)
关键词:
微小核糖核酸-29; 慢性盆腔炎; 炎症因子; Toll 样受体4/ 核因子κB
分类号:
R-332；R711.33
DOI:
10.3969/j.issn.1671-7414.2022.06.003
文献标志码:
A
摘要:
目的 探究微小核糖核酸(micro RNA,miR)-29 对大鼠慢性盆腔炎模型的作用并探究其作用机制。方法 将80 只SPF 级SD 大鼠随机分为四组，分别为对照组(n=20)、模型组(n=20)、阴性对照组(n=20)和试验组(n=20)。模型组、阴性对照组和试验组通过机械损伤及接种混合菌构建大鼠慢性盆腔炎模型，阴性对照组和试验组造模后通过尾静脉注射5nmol 阴性对照(negative control, NC)antagomiR 和miR-29 antagomiR。HE 染色检测大鼠子宫组织病理特性；酶联免疫吸附测定(enzyme linked immunosorbent assay，ELISA)法检测各组大鼠外周血炎症相关指标：白细胞介素(interleukin，IL)-1β，白细胞介素(IL)-6，白细胞介素(IL)-8 和肿瘤坏死因子(tumor necrosis factor，TNF)-α 水平；实时荧光定量PCR(quantitative real-time PCR，qPCR)检测子宫组织miR-29，Toll 样受体4(Toll-like receptors，TLR)4，核因子κB(nuclear factor，NF-κB)及NK-κB 抑制蛋白α(IκB α)表达水平；WesternBlot 检测子宫组织TLR4，NF-κB 和IκB α 蛋白表达水平。结果 HE 染色结果表明，与对照组相比，模型组、阴性对照组和试验组大鼠出现明显慢性盆腔炎症状，试验组子宫组织损伤显著缓解；ELISA 结果表明：模型组大鼠外周血IL-1β，IL-6，IL-8 和TNF-α 水平显著高于对照组，与模型组和阴性对照组相比，试验组炎症因子IL-1β，IL-6，IL-8 和TNF-α 释放显著降低，差异有统计学意义(t=3.021 ～ 6.878，均P ＜ 0.05)；qPCR 结果表明，与对照组相比，模型组miR-29，TLR4，NF-κB 和IκB α 的mRNA 表达显著上升，与模型组和阴性对照组相比，试验组miR-29 ，TLR4，NF-κB 和IκB α的 mRNA 表达显著降低，差异有统计学意义(t=3.917 ～ 8.095，均P ＜ 0.05)。Western Blot 结果表明：与对照组相比，模型组TLR4，NF-κB 和IκB α 蛋白表达显著上升，与模型组和阴性对照组相比，试验组TLR4，NF-κB 和IκBα 蛋白表达显著降低，差异有统计学意义(t=2.987 ～ 8.045，均P ＜ 0.05)。结论 慢性盆腔炎模型大鼠子宫组织中miR-29 显著高表达，下调miR-29 具有对大鼠子宫的保护作用，其机制可能与抑制炎症因子释放及TLR4/NF-κB 信号通路的调控相关。
Abstract:
Objective To explore the effect of miR-29 on chronic pelvic inflammatory disease model in rats and its mechanism. Methods 80 SPF SD rats were randomly divided into 4 groups: control group (n=20), model group (n=20), NC group (n= 20) and experimental group (n=20). The model group, NC group and experimental group were established with mechanical injury and inoculation of mixed bacteria. After modeling, the NC group and experimental group were injected with 5 nmol NC antagomiR and miR-29 antagomiR through caudal vein. HE staining was used to detect the case characteristics of rat uterine tissue, and ELISA was used to detect the peripheral blood inflammation related index interleukin-1-β(IL-1β)，interleukin-6 (IL-6)， interleukin-8 (IL-8) and tumor necrosis factor α(TNF-α). The levels of miR-29, Toll like receptor 4 (TLR4) and nuclear factor were detected by qPCR nuclear factorκB(NF-κB) and NK-κB inhibitor protein α(IκBα). The expression levels of TLR4 and NF-κB and IκB α protein expression level in uterine tissues were detected by Western blot. Results The results of HE staining showed that compared with the control group, the rats in the model group, NC group and experimental group had obvious symptoms of chronic pelvic inflammatory disease, and the uterine tissue injury in the experimental group was significantly relieved. The results of ELISA showed that the peripheral blood IL-1β, IL-6, IL-8 and TNF-α in the model group was significantly reduced. Compared with the model group and NC group, the level of inflammatory factor IL-1β ，IL-6， IL-8 and TNF-α in the experimental group was significantly higher than that in the control group. The release of miR-29, TLR4, NF-κB and IκB α in the model group was significantly lower than that in the control group, the differences were statistically significant(t = 3.021 ~ 6.878, all P ＜ 0.05). The mRNA expression of miR-29, TLR4, NF-κB and IκB α in the experimental group increased significantly compared with the model group and NC group.The mRNA expression of TLR4 and NF-κB and IκB α in the model group was significantly lower than that in the control group, the differences were statistically significant(t = 3.917 ~ 8.095, all P ＜ 0.05) .The expression of TLR4, NF-κB and IκB α in the experimental group increased significantly compared with the model group and NC group. The protein expression decreased significantly, the differences were statistically significant(t = 2.987 ~ 8.045, all P ＜ 0.05). Conclusion MiR-29 was significantly overexpressed in the uterus of Rats with chronic pelvic inflammatory disease. Down regulation of miR-29 had a protective effect on the uterus of rats, and its mechanism may be related to the inhibition of inflammatory factor release and TLR4/NF-κB，the regulation of signaling pathway is related.

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备注/Memo

备注/Memo:
基金项目：河北省医学科学研究课题 (20201609)。
作者简介：马慧敏(1981-)，女，本科，主治医师，研究方向：妇产科学。
通讯作者：金瑞林 (1971-)，女，硕士研究生，主治医师，研究方向：妇产科学，E-mail:dpdwionmvahzrdv1@21cn.com。
更新日期/Last Update: 2022-11-15