Acute Respiratory Distress Syndrome

OIP5-AS1[LncRNA]

miR-223[miRNA]

NLRP3[mRNA]

The Serum Of Ali/Ards Patients

Homo sapiens (human)

J Gene Med. 2021 Aug 4:e3385. doi: 10.1002/jgm.3385.

LncRNA OIP5-AS1 knockdown or miR-223 overexpression can alleviate LPS-induced ALI/ARDS by interfering with miR-223/NLRP3-mediated pyroptosis.

Dual-luciferase reporter assay;ELISA;MTT assay;Western blot;Flow Cytometry assay;Luciferase reporter assay;MTT assay;

BACKGROUND: Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are life-threatening diseases and endothelial barrier injury is an important contributor to the pathogenesis of ALI/ARDS. LncRNA has been proved to participate in the progression of ALI/ARDS. Our study aimed to investigate the function of lncRNA OIP5-AS1 in LPS-induced ALI/ARDS. METHODS: OIP5-AS1 and miR-223 levels were detected by PCR in the serum of ALI/ARDS patients or healthy donors. MTT assay were performed to detect the proliferation of HPMECs. Flow cytometry were performed to detect the apoptosis of HPMECs. The protein levels of NLRP3, ASC, GSDMD-N, caspase-1 were measured by western blot to detect the pyroptosis of HPMECs. IL-1b, IL-6, IL-18 and IL-10 was detected by ELISA to measure the inflammatory response of HPMECs. And production of ROS, SOD and MDA was measured to determine the oxidative stress of HPMECs. Targets of OIP5-AS1 and miR-223 were predicted by StarBase and confirmed by dual-luciferase reporter assay. RESULTS: We found that OIP5-AS1 was upregulated, while miR-223 was downregulated in the serum of ALI/ARDS patients and LPS-treated HPMECs. Functionally, knockdown of OIP5-AS1 induced proliferation and inhibited apoptosis, pyroptosis, inflammatory response and oxidative stress of LPS-treated HPMECs. Interestingly, miR-223 was a target of OIP5-AS1 and miR-223 inhibition abolished the effects of si-OIP5-AS1 on LPS-induced HPMECs. More importantly, miR-223 directly targeted NLRP3, miR-223 overexpression also promoted proliferation and inhibited apoptosis, pyroptosis, inflammatory response and oxidative stress of LPS-treated HPMECs and which was abolished by NLRP3 overexpression. Finally, we found that OIP5-AS1 knockdown and miR-223 overexpression could both alleviate LPS-induced ALI/ARDS in vivo. CONCLUSION: Together, we find that LncRNA OIP5-AS1 aggravates LPS-induced ALI/ARDS via miR-223/NLRP3 axis and provides new targets for ALI/ARDS therapy.