| 摘要: |
| 目的：探讨微小RNA（miR）-30a-5p调控白细胞介素17A（IL17A）影响脂多糖诱导RAW264.7细胞炎症反应的作用机制。方法：将RAW264.7细胞分为control组、脂多糖组、miR-30a-5p过表达对照组、miR-30a-5p过表达组、过表达miR-30a-5p+IL17A对照组、过表达miR-30a-5p+IL17A组，除control组外其余组RAW264.7细胞均采用脂多糖诱导，并进行相应的质粒转染。EdU测定细胞增殖；TUNEL测定细胞凋亡；试剂盒测定细胞上清液中NO、IL-1β、IL-6和TNF-α的含量；qRT-PCR检测miR-30a-5p和IL17A mRNA水平；Western blot检测IL17A蛋白表达；双荧光素酶报告基因分析miR-30a-5p与IL17A的靶向关系。结果：与control组相比，脂多糖组RAW264.7细胞EdU阳性率、miR-30a-5p水平降低，TUNEL阳性率、NO、IL-1β、IL-6和TNF-α含量、IL17A mRNA及其蛋白水平均提高（P<0.05）；与miR-30a-5p过表达对照组相比，miR-30a-5p过表达组RAW264.7细胞EdU阳性率、miR-30a-5p水平提高，TUNEL阳性率、NO、IL-1β、IL-6和TNF-α含量、IL17A mRNA及其蛋白水平均降低（P<0.05）；与过表达miR-30a-5p+IL17A对照组相比，过表达miR-30a-5p+IL17A组RAW264.7细胞EdU阳性率降低，TUNEL阳性率、NO、IL-1β、IL-6和TNF-α含量、IL17A mRNA及其蛋白水平提高（P<0.05）；双荧光素酶报告基因分析得出，miR-30a-5p与IL17A之间具有靶向关系（P<0.05）。结论：miR-30a-5p通过抑制IL17A表达减轻脂多糖诱导RAW264.7细胞的炎症反应。 |
| 关键词: miR-30a-5p IL17A RAW264.7细胞 脂多糖 炎症反应 |
| DOI： |
| 分类号: |
| 基金项目:浙江省市县级科技计划项目 |
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| The mechanism by which miR-30a-5p affects lipopolysaccharide induced inflammatory response in RAW264.7 cells by regulating IL17A |
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朱鹏1,2,3, xie yue qun1,2,3, shao long long1,2,3
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1.Wenzhou People'2.'3.s Hospital
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| Abstract: |
| Objective: To discuss the mechanism by which microRNA (miR)-30a-5p affects lipopolysaccharide induced inflammatory response in RAW264.7 cells by regulating interleukin 17A (IL17A). Methods: RAW264.7 cells were assigned into control group, lipopolysaccharide group, miR-30a-5p overexpression control group, miR-30a-5p overexpression group, miR-30a-5p overexpression+IL17A control group, and miR-30a-5p overexpression+IL17A group. Except for the control group, RAW264.7 cells in all other groups were induced with lipopolysaccharide and transfected with corresponding plasmids. EdU was performed to measure cell proliferation. TUNEL was performed to measure cell apoptosis. The reagent kits were used to measure the levels of NO, IL-1β, IL-6, and TNF-α in the cell supernatant. QRT-PCR was performed to detect the levels of miR-30a-5p and IL17A mRNA. Western blot was used to discuss the expression of IL17A protein. Moreover, dual luciferase reporter gene analysis was performed to discuss the targeting relationship between miR-30a-5p and IL17A. Results: Compared with the control group, the lipopolysaccharide group showed a decrease in EdU positivity rate and miR-30a-5p level in RAW264.7 cells, while an increase in TUNEL positivity rate, NO, IL-1β, IL-6, TNF-α contents, IL17A mRNA and protein levels (P<0.05). Compared with the miR-30a-5p overexpression control group, the miR-30a-5p overexpression group showed an increase in EdU positivity rate and miR-30a-5p level in RAW264.7 cells, while a decrease in TUNEL positivity rate, NO, IL-1β, IL-6, TNF-α contents, IL17A mRNA and protein levels (P<0.05). Compared with the miR-30a-5p overexpression+IL17A control group, the miR-30a-5p overexpression+IL17A group showed a decrease in EdU positivity rate in RAW264.7 cells, while an increase in TUNEL positivity rate, NO, IL-1β, IL-6, TNF-α contents, IL17A mRNA and protein levels (P<0.05). The dual luciferase reporter gene analysis showed a targeted relationship between miR-30a-5p and IL17A (P<0.05). Conclusion: MiR-30a-5p alleviates lipopolysaccharide induced inflammatory response in RAW264.7 cells by inhibiting IL17A expression. |
| Key words: miR-30a-5p IL17A RAW264.7 cells lipopolysaccharide inflammatory response |