维生素D3抑制博莱霉素诱导的肺部炎症反应以及肺上皮—间质转化

发布时间：2018-07-30 07:56

【摘要】：目的:早期肺部炎症和肺上皮间质转化(EMT)在肺纤维化过程起重要作用。越来越多的证据表明,骨化三醇(即活化型维生素D3)具有抗炎作用。本研究的目的是调查维生素D3对博来霉素诱导的早期肺部炎症和随后EMT的抑制效应。方法:成年C57BL/6J雄性小鼠(8周,24-26g),随机分为PBS对照组(Control),BLM模型组(BLM),不同剂量维生素D3保护组(BLM+Vit D)。对照组给予气管插管注射PBS,单纯BLM组小鼠气管内插管注射BLM(3.0毫克/千克),不同剂量维生素D3保护组小鼠腹腔内(i.p)注射不同剂量维生素D3(0.2、1.0或5.0 mg/kg)予以预处理。维生素D3预处理时间均为给予LPS前48 h、24 h、1h。BLM注射后24小时,7天和14天,取肺组织。部分肺脏组织经4%多聚甲醛固定后制备石蜡包埋组织切片,用HE染色法观察病理学改变,用Western Blot检测肺组织p Akt/Akt,pp38,NF-κB p65水平,通过免疫组化检测A-SMA和NF-κB p65水平,RT-PCR检测各种细胞炎性因子表达。结果:维生素D3减轻BLM诱导的肺部炎症细胞浸润。BLM诱导肺部炎性细胞因子也被维生素D3抑制。,维生素D3减少BLM诱导NF-k B p65入核、减轻肺p38 MAPK和Akt磷酸化。进一步分析表明,BLM诱导的平滑肌肌动蛋白(a-SMA也被维生素D3抑制。维生素D3抑制BLM诱导的转化生长因子1(TGF-b1)上调和Smad磷酸化。结论:维生素D3抑制BLM-诱导的早期肺部炎症和随后的EMT转化。
[Abstract]:Objective: early pulmonary inflammation and pulmonary epithelial interstitial transformation (EMT) play an important role in the process of pulmonary fibrosis. There is growing evidence that calcitriol, or activated vitamin D 3, has anti-inflammatory effects. The aim of this study was to investigate the inhibitory effects of vitamin D 3 on bleomycin induced early pulmonary inflammation and subsequent EMT. Methods: adult C57BL/6J male mice (8-week-24 ~ 26g) were randomly divided into PBS control group (Control) / BLM model group), (BLM), treated group with different doses of vitamin D _ 3 (BLM Vit D). The control group was injected with endotracheal intubation, the mice of BLM group were injected with BLM (3.0 mg / kg), and the mice of different doses of vitamin D _ 3 (i.p) were pretreated with different doses of vitamin D _ 3 (0.2g 1.0 or 5.0 mg/kg). The pretreatment time of vitamin D _ 3 was 24 hours, 7 days and 14 days after administration of LPS. Paraffin-embedded tissue sections were prepared from some lung tissues fixed with 4% paraformaldehyde. Pathological changes were observed by HE staining, and the level of p65 in p Akt / AktP38 and NF- 魏 B p65 in lung tissue was detected by Western Blot. The levels of A-SMA and NF- 魏 B p65 were detected by immunohistochemistry and RT-PCR was used to detect the expression of various cytokines. Results: vitamin D _ 3 alleviated the infiltration of pulmonary inflammatory cells induced by BLM. BLM-induced pulmonary inflammatory cytokines were also inhibited by vitamin D _ 3. Vitamin D _ 3 decreased BLM induced NF-k B p65 entry into the nucleus and reduced p38 MAPK and Akt phosphorylation. Further analysis showed that smooth muscle actin (a-SMA) induced by BLM was also inhibited by vitamin D 3. Vitamin D 3 inhibits BLM induced up-regulation of transforming growth factor 1 (TGF-b1) and Smad phosphorylation. Conclusion: vitamin D 3 inhibits early lung inflammation induced by BLM- and subsequent EMT transformation.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R563

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