| 摘要: |
| 目的:探讨五味子苷B(Sch B)在幼年肺炎小鼠的作用机制。方法:将90 只幼年雄性小鼠随机分为对照组、模型组、Sch B
低剂量组(Sch BL组)、Sch B 高剂量组(Sch BH组)、Sch BH+antagomir-NC 组、Sch BH+miR-370-3p antagomir组各15 只。Sch BL
组和Sch BH 组小鼠分别灌胃20、60 mg/ kg 的Sch B;Sch BH+antagomir-NC 组和Sch BH+miR-370-3p antagomir组,先将1 nmol 的
miR-370-3p antagomir、antagomir-NC 用20 μL 的磷酸盐缓冲液(PBS)溶解,在Sch B 灌胃后将miR-370-3p antagomir、antagomir-NC
质粒分别经尾静脉注射到小鼠体内;对照组和模型组给予等量生理盐水。每天1 次,持续给药7 d。测定各组小鼠肺组织的湿
干质量比;采用苏木精-伊红(HE)染色观察各组小鼠肺组织的病理形态;检测各组小鼠肺组织中炎症因子水平;流式细胞术检
测外周血T 淋巴细胞亚群;实时定量反转录聚合酶链式反应(qRT-PCR)检测肺组织中miR-370-3p 和CCL3 的mRNA 水平;双荧
光素酶实验验证miR-370-3p 与CCL3 的靶向关系。结果:与对照组相比,模型组幼鼠肺组织结构紊乱,肺泡壁变厚,出现大量炎
性细胞浸润,组织受损严重,肺组织湿干质量比、CD8+ 、肿瘤坏死因子(TNF)-α、白细胞介素( IL)-6、CCL3 mRNA 水平均升高,
CD4+ 、CD4+/CD8+ 、miR-370-3p 水平均降低(P 均<0.05)。与模型组相比,Sch BL 组和Sch BH 组幼鼠肺组织中肺泡壁变薄,炎
性细胞浸润明显减少,损伤减轻,肺组织湿干质量比、CD8+、TNF-α、IL-6、CCL3 mRNA 水平均降低,CD4+、CD4+ / CD8+、miR-370-3p
水平均升高(P 均<0. 05)。加入miR-370-3p antagomir 进行回补实验,结果显示Sch B 对肺炎幼鼠免疫功能和炎症保护作用被
逆转,且CCL3 mRNA 水平升高(P<0. 05);双荧光素酶报告基因实验验证了miR-370-3p 与CCL3 存在靶向关系。结论:Sch B 能
够通过靶向调节miR-370-3p/ CCL3 轴来增强幼年肺炎小鼠免疫功能,并抑制炎症反应。 |
| 关键词: 五味子苷B miR-370-3p/CCL3轴 肺炎 免疫功能 炎症 |
| DOI：doi:10.13407/j.cnki.jpp.1672-108X.2024.05.001 |
|
| 基金项目:山东省中医药科技发展计划项目,编号2019-0031。 |
|
| Impacts of Schisandrin B on Immune Inflammatory Factors in Juvenile Mice with Pneumonia by RegulatingmiR-370-3p/CCL3 Axis |
| Yang Cuiling1, Gao Junming2, Dong Lijun2, Wang Limin3, Zhang Yongfeng4 |
| (1. School of Clinical Medicine, Shandong
Second Medical University, Shandong Weifang 250000, China; 2. Anqiu People’s Hospital, Shandong Weifang 262199, China; 3.
School of Pharmacy, Shandong University of Traditional Chinese Medicine, Jinan 250355, China; 4. Affiliated Hospital of Weifang
Medical College, Shandong Weifang 261035, China) |
| Abstract: |
| Objective: To investigate the mechanism of Schisandrin B (Sch B) in juvenile mice with pneumonia. Methods: Ninety
juvenile male mice were randomly divided into the control group, model group, Sch B low dose group (Sch BL group), Sch B high dose
group (Sch BH group), Sch BH+ antagomir-NC group and Sch BH+miR-370-3p antagomir group, with 15 mice in each group. Sch BL
group and Sch BH group were given 20 and 60 mg/ kg Sch B, respectively. For the Sch BH+antagomir-NC group and Sch BH+miR-370-
3p antagomir group, 1 nmol of miR-370-3p antagomir and antagomir-NC were first dissolved in 20 μL of phosphate buffer solution
(PBS), and Sch B was gavaged, miR-370-3p antagomir, antagomir-NC plasmid were injected into mice by tail vein, respectively. The
control group and model group were given the same amount of normal saline. The drug was administered once a day for 7 d. The wet to
dry weight ratio of pulmonary tissue of mice in each group was measured. Hematoxylin Eosin (HE) staining was applied to observe the
pathological morphology of pulmonary tissue in each group of mice. Levels of inflammatory factors in pulmonary tissues of mice were
detected. Flow cytometry was applied to detect T lymphocyte subsets in peripheral blood. Quantitative real time polymerase chain
reaction (qRT-PCR) was used to detect the levels of miR-370-3p and CCL3 mRNA in the pulmonary tissues. Double Luciferase
experiment was applied to verify the targeting relationship between miR-370-3p and CCL3. Results: Compared with the control group,
the pulmonary tissue structure of juvenile mice in the model group was disordered, with thickened alveolar walls, a large number of
inflammatory cell infiltration, and severe tissue damage, the value of wet to dry weight, percentage of CD8+ , levels of tumour necrosises
factor (TNF)-α, interleukin (IL)-6 and CCL3 mRNA increased, CD4+ ,CD4+ / CD8+ , and miR-370-3p decreased (P < 0. 05).
Compared with the model group, the alveolar wall in pulmonary tissue of juvenile mice in Sch BL group and Sch BH group became
thinner, inflammatory cell infiltration reduced significantly, and damage was alleviated, the value of wet to dry weight, CD8+ , levels of
TNF-α, IL-6 and CCL3 mRNA decreased, CD4+ , CD4+ / CD8+ , and miR-370-3p increased (P<0. 05). The addition of miR-370-3p
antagomir for compensatory experiments showed that the protective effects of Sch B on immune function and inflammation in juvenile mice with pneumonia were reversed, and the level of CCL3 mRNA increased (P<0. 05). Double Luciferase reporter gene experiment verified
that there was a targeting relationship between miR-370-3p and CCL3. Conclusion: Sch B can enhance the immune function of juvenilea
mice with pneumonia and inhibit the inflammatory response by targeting the miR-370-3p/ CCL3 axis. |
| Key words: Schisandrin B miR-370-3p/ CCL3 axis pneumonia immune function inflammation |
