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| Protective Effect of miR-29a on Lipopolysaccharide-induced Human Pulmonary Microvascular Endothelial Cells Injury by Targeting PTEN Expression |
CHEN Yu-qiong1,2,TAN Wen-hua2,LIU Hai-feng1,CHEN Gen1,2,**( ) |
1 The First People’s Hospital of Chenzhou, Chenzhou 423000, China
2 Institute of Pharmacy and Pharmacology, University of South China, Hengyang 421001, China |
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Abstract Aim: To investigate the effect of miR-29a on lipopolysaccharide-induced injury in human pulmonary microvascular endothelial cells and its potential mechanism. Methods: The LPS-induced HPMVECs injury model was constructed. The expression level of miR-29a was detected by RT-qPCR. The concentration of LDH was measured by ELISA. MTT assay was used to detect the cell proliferation. The flow cytometry was applied to determine the HPMVECs apoptosis. The protein levels of PTEN, p-Akt, Akt, p-FOXO3a, FOXO3a and Bim were determined by Western blot. The targeting relationship between miR-29a and PTEN was predicted by Microcosm, starBase, Pictar, TargetScan and confirmed by luciferase test. Results: LPS treatment of HPMVECs significantly reduced the expression level of miR-29a and cell viability,induced the increase of LDH release amount and cell apoptic rate, upregulated the expression of PTEN and Bim protein, and down regulated the expression of p-Akt/Akt and p-FOXO3a/FOXO3a (P<0.05). Overexpression of miR-29a reversed the injury of LPS to HPMVECs. Dual luciferase reporter gene assay confirmed that PTEN was a negative regulatory target gene of miR-29a. The protein expression of PTEN was significantly down regulated by miR-29a mimics, and up-regulated by miR-29a inhibitors(P<0.05). However, the expression level of PTEN mRNA had no statistically significant difference (P>0.05). Conclusion: Overexpression of miR-29a, which targets inhibition of PTEN protein expression, protects against LPS-induced HUVECs injury by activating the Akt/FOXO3a/Bim pathway.
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Received: 22 December 2020
Published: 01 June 2021
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Corresponding Authors:
Gen CHEN
E-mail: myth3878@aliyun.com
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