Shi Mengmeng, Li Qi, Zhou Xiangdong, Huang Huaping, Wang Jie
Department of Respiratory Medicine, Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China.
Department of Respiratory Medicine, Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010; Department of Respiratory Meddicine, Affiliated Hospital of Hainan Medical College, Haikou 570100, China.
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2016 Jan;41(1):1-8. doi: 10.11817/j.issn.1672-7347.2016.01.001.
To explore the role NDRG2/Zc3h12d in the regulation of neutrophil nuclear transcription factor-κB signaling pathway and the underlying mechanisms.
Sixteen HBE cells were incubated with lipopolysaccharide (LPS) to establish airway mucus hypersecretion model, which was transfected with NDRG2 or Zc3h12d siRNA. The cells were divided into 5 groups: a LPS+NDRG2 siRNA (Group A), a LPS+ NDRG2 and Zc3h12d siRNAs (Group B), a LPS+Zc3h12d siRNA (Group C), a LPS+ empty plasmid (Group D), and a negative control group (Group E). The reverse transcription-polymerase chain reaction (RT-PCR) was used to measure the expression level of mucin (MUC) 5AC mRNA. The levels of MUC5AC and the inflammatory factors were examined by the enzyme-linked immunosorbent assay (ELISA). The NF-κB p65 and the Zc3h12d protein levels were measured by Western blot. The MUC5C expression was further examined by laser confocal method.
Compared with Group B, the levels of MUC5AC mRNA and protein in Group A were decreased (P<0.05), there was no significant difference in the MUC5AC mRNA and MUC5AC protein levels between the Group B and the Group C (P>0.05). Compared with Group D, the MUC5AC (mRNA and protein) and inflammatory factor levels in the Group A were significantly decreased (P<0.05); compared with Group E, after incubation with LPS, the levels of MUC5AC (mRNA and protein) and inflammatory factors in the Group D was increased, with significantly difference (P<0.05).
NDRG2 can regulate the function of NF-κB signaling pathway through the deubiquitylating enzyme of Zc3h12d.
探讨NDRG2/Zc3h12d在调节中性粒细胞核转录因子-κB信号通路中的作用及其潜在机制。
将16个HBE细胞与脂多糖（LPS）孵育以建立气道黏液高分泌模型，并用NDRG2或Zc3h12d小干扰RNA（siRNA）转染。细胞分为5组：LPS+NDRG2 siRNA组（A组）、LPS+NDRG2和Zc3h12d siRNAs组（B组）、LPS+Zc3h12d siRNA组（C组）、LPS+空质粒组（D组）和阴性对照组（E组）。采用逆转录-聚合酶链反应（RT-PCR）检测黏蛋白（MUC）5AC mRNA的表达水平。采用酶联免疫吸附测定（ELISA）检测MUC5AC和炎症因子水平。采用蛋白质免疫印迹法检测NF-κB p65和Zc3h12d蛋白水平。采用激光共聚焦法进一步检测MUC5C表达。
与B组比较，A组MUC5AC mRNA和蛋白水平降低（P<0.05），B组和C组MUC5AC mRNA和MUC5AC蛋白水平差异无统计学意义（P>0.05）。与D组比较，A组MUC5AC（mRNA和蛋白）和炎症因子水平显著降低（P<0.05）；与E组比较，LPS孵育后，D组MUC5AC（mRNA和蛋白）和炎症因子水平升高，差异有统计学意义（P<0.05）。
NDRG2可通过Zc3h12d去泛素化酶调节NF-κB信号通路的功能。
