摘要
目的探讨干扰LncRNA PVT1对脂多糖(LPS)所致人支气管上皮细胞16HBE炎症、氧化应激、凋亡的影响及其可能作用机制。方法采用LPS诱导16HBE细胞建立细胞损伤模型,将16HBE细胞分为对照组Control组、LPS组、LPS+si-NC组、LPS+si-PVT1组、LPS+si-PVT1+anti-miR-NC组、LPS+si-PVT1+anti-miR-1301-3p组;采用qRT-PCR检测PVT1、miR-1301-3p的表达量;采用ELISA法检测白细胞介素6(IL-6)、白细胞介素13(IL-13)、肿瘤坏死因子-α(TNF-α)的水平;采用试剂盒检测超氧化物歧化酶(SOD)、丙二醛(MDA)、乳酸脱氢酶(LDH)的含量;采用流式细胞术检测细胞凋亡率;双荧光素酶报告实验检测PVT1与miR-1301-3p的靶向关系。结果与Control组比较,LPS组PVT1的表达水平升高(P<0.05),miR-1301-3p的表达水平降低(P<0.05),IL-6、IL-13、TNF-α的水平与MDA、LDH的含量及凋亡率升高(P<0.05),SOD的含量降低(P<0.05);转染siPVT1后可明显降低IL-6、IL-13、TNF-α的水平与MDA、LDH的含量及凋亡率(P<0.05),提高SOD的含量(P<0.05);双荧光素酶报告实验证实PVT1可靶向调控miR-1301-3p;共转染si-PVT1与anti-miR-1301-3p可明显逆转si-PVT1对LPS所致16HBE细胞炎症、氧化应激及凋亡的作用。结论干扰PVT1可通过上调miR-1301-3p而抑制炎症反应、氧化应激及细胞凋亡从而减轻LPS所致支气管上皮细胞损伤。
This study was designed to explore the effect of LncRNA PVT1 interference on the inflammation,oxidative stress,and apoptosis of human bronchial epithelial cells induced by lipopolysaccharide(LPS)and its possible mechanism.LPS was used to induce 16HBE cells for establishing a cell injury model.16HBE cells were divided into control group,LPS group,LPS+si-NC group,LPS+si-PVT1 group,LPS+si-PVT1+anti-miR-NC group,and LPS+si-PVT1+anti-miR-1301-3p group.qRT-PCR was used to detect the expression of PVT1 and miR1301-3p;ELISA was sued to detect the levels of IL-6,IL-13 and TNF-α.Commercial kits were used to detect the content of SOD,MDA and LDH;flow cytometry was used to detect the apoptosis rate.The dual luciferase reporter experiment was carried out to detect the targeting relationship between PVT1 and miR-1301-3p.Compared with the control group,the levels of PVT1,IL-6,IL-13 and TNF-α,the contents of MDA,LDH,as well as the apoptosis rate in the LPS group were increased(P<0.05),while the miR-1301-3p level and SOD content were decreased(P<0.05).si-PVT1 transfection could significantly revere these changes mentioned above(P<0.05).Furthermore,the dual luciferase report experiment confirmed that PVT1 could target and regulate miR-1301-3p;co-transfection of siPVT1 and anti-miR-1301-3p could significantly antagonise the effects of si-PVT1 transfection on LPS-induced inflammation,oxidative stress and apoptosis of 16HBE cells.In conclusion,PVT1 interference could inhibit the inflammation,oxidative stress and apoptosis of 16HBE cells by up-regulating miR-1301-3p,thereby reducing LPS-induced damage of bronchial epithelial cells.
作者
颜维孝
宋亚娣
李娟
YAN Weixiao;SONG Yadi;LI Juan(Department of Critical Medicine,Qinghai Provincial Women and Children's Hospital,Xining 810007,China;Department of Respiratory Medicine,Qinghai Provincial Women and Children's Hospital,Xining 810007,China)
出处
《免疫学杂志》
CAS
CSCD
北大核心
2022年第1期74-80,共7页
Immunological Journal
