Down-regulation of miR-199a-5p inhibits lipopolysaccharide- induced apoptosis of rat type Ⅱ alveolar epithelial cell line AECⅡ
Objective To investigate the expression of miR-199a-5p in the serum of patients with chronic obstructive pulmonary disease (COPD) and its effect on lipopolysaccharide (LPS)-induced apoptosis of rat type Ⅱ alveolar epithelial cells (AECⅡ) and possible mechanism. Methods Forty-five patients with COP...
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| Format: | Article |
| Language: | zho |
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Institute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College.
2021-04-01
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| Series: | Jichu yixue yu linchuang |
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| Online Access: | http://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/a200462.pdf |
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| author | WANG Wei, ZHAO Si-lin, FAN Fu-yuan, JIN Zhao-hui, LI Da, FU Yan |
| author_facet | WANG Wei, ZHAO Si-lin, FAN Fu-yuan, JIN Zhao-hui, LI Da, FU Yan |
| author_sort | WANG Wei, ZHAO Si-lin, FAN Fu-yuan, JIN Zhao-hui, LI Da, FU Yan |
| collection | DOAJ |
| description | Objective To investigate the expression of miR-199a-5p in the serum of patients with chronic obstructive pulmonary disease (COPD) and its effect on lipopolysaccharide (LPS)-induced apoptosis of rat type Ⅱ alveolar epithelial cells (AECⅡ) and possible mechanism. Methods Forty-five patients with COPD and 45 controls were enrolled in the first affiliated hospital of Hunan University of Traditional Chinese Medicine from April 2017 to December 2018 . AEC Ⅱ cells were cultured in vitro and were divided into control group , LPS group, anti-miR- 199a-5p+ LPS group, anti-miR-NC+LP group,anti-miR-199a-5p+si-FZD4+Lgroup and anti-miR-199a-5p+si-NC+LPS group. RT-qPCR was used to detect the expression of miR-199a-5p and FZD4 mRNA in serum and cells, flow cytometry was used to detect the apoptosis rate and Western blot was used to detect FZD4, B-lymphoma-2 (Bcl-2) and B-lymphoma-2 related proteins (Bax) in cells. The dual luciferase reporter gene experiment verified the targeting relationship between miR-199a-5p and FZD4. Results Compared with the control, the expression of miR-199a-5p in serum of COPD patients significantly increased (P＜0.05), and the expression of FZD4 mRNA significantly reduced (P＜0.05). Compared with the control group, the expression of miR-199a-5p in the LPS group significantly increased(P＜0.05), the mRNA and protein expression of FZD4 significantly reduced (P＜0.05), the apoptosis rate and the expression level of Bax protein significantly increased (P＜0.05), and the expression of Bcl-2 protein significantly reduced (P＜0.05). Compared with anti-miR-NC+LPS group, cell apoptosis and the expression of Bax protein in the anti-miR-199a-5p+LPS group significantly reduced (P＜0.05), while the expression of Bcl-2 protein significantly increased (P＜ 0.05). There was no significant change in the detection indexes between the LPS group and the anti-miR-NC+LPS group (P＞0.05). miR-199a-5p targets FZD4, compared with anti-miR-199a-5p+si-NC+LPS group, cell apoptosis rate and the expression level of Bax protein in the anti-miR-199a-5p+ si-FZD4+LPS group all significantly increased (P＜0.05), while the expression of Bcl-2 protein significantly reduced (P＜0.05). Conclusions The expression of miR-199a-5p is increased in serum of COPD patients and in LPS-induced AECⅡcells. Down-regulating miR-199a-5p expression may inhibit LPS-induced AECⅡ cell apoptosis by up-regulating FZD4 expression. |
| first_indexed | 2024-03-08T16:53:33Z |
| format | Article |
| id | doaj.art-00ff5aa182eb4f4b965130e1768b60b5 |
| institution | Directory Open Access Journal |
| issn | 1001-6325 |
| language | zho |
| last_indexed | 2024-03-08T16:53:33Z |
| publishDate | 2021-04-01 |
| publisher | Institute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College. |
| record_format | Article |
| series | Jichu yixue yu linchuang |
| spelling | doaj.art-00ff5aa182eb4f4b965130e1768b60b52024-01-05T03:04:57ZzhoInstitute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College.Jichu yixue yu linchuang1001-63252021-04-01414521527Down-regulation of miR-199a-5p inhibits lipopolysaccharide- induced apoptosis of rat type Ⅱ alveolar epithelial cell line AECⅡWANG Wei, ZHAO Si-lin, FAN Fu-yuan, JIN Zhao-hui, LI Da, FU Yan0Department of Respiratory Medicine, the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine, Changsha 410007, ChinaObjective To investigate the expression of miR-199a-5p in the serum of patients with chronic obstructive pulmonary disease (COPD) and its effect on lipopolysaccharide (LPS)-induced apoptosis of rat type Ⅱ alveolar epithelial cells (AECⅡ) and possible mechanism. Methods Forty-five patients with COPD and 45 controls were enrolled in the first affiliated hospital of Hunan University of Traditional Chinese Medicine from April 2017 to December 2018 . AEC Ⅱ cells were cultured in vitro and were divided into control group , LPS group, anti-miR- 199a-5p+ LPS group, anti-miR-NC+LP group,anti-miR-199a-5p+si-FZD4+Lgroup and anti-miR-199a-5p+si-NC+LPS group. RT-qPCR was used to detect the expression of miR-199a-5p and FZD4 mRNA in serum and cells, flow cytometry was used to detect the apoptosis rate and Western blot was used to detect FZD4, B-lymphoma-2 (Bcl-2) and B-lymphoma-2 related proteins (Bax) in cells. The dual luciferase reporter gene experiment verified the targeting relationship between miR-199a-5p and FZD4. Results Compared with the control, the expression of miR-199a-5p in serum of COPD patients significantly increased (P＜0.05), and the expression of FZD4 mRNA significantly reduced (P＜0.05). Compared with the control group, the expression of miR-199a-5p in the LPS group significantly increased(P＜0.05), the mRNA and protein expression of FZD4 significantly reduced (P＜0.05), the apoptosis rate and the expression level of Bax protein significantly increased (P＜0.05), and the expression of Bcl-2 protein significantly reduced (P＜0.05). Compared with anti-miR-NC+LPS group, cell apoptosis and the expression of Bax protein in the anti-miR-199a-5p+LPS group significantly reduced (P＜0.05), while the expression of Bcl-2 protein significantly increased (P＜ 0.05). There was no significant change in the detection indexes between the LPS group and the anti-miR-NC+LPS group (P＞0.05). miR-199a-5p targets FZD4, compared with anti-miR-199a-5p+si-NC+LPS group, cell apoptosis rate and the expression level of Bax protein in the anti-miR-199a-5p+ si-FZD4+LPS group all significantly increased (P＜0.05), while the expression of Bcl-2 protein significantly reduced (P＜0.05). Conclusions The expression of miR-199a-5p is increased in serum of COPD patients and in LPS-induced AECⅡcells. Down-regulating miR-199a-5p expression may inhibit LPS-induced AECⅡ cell apoptosis by up-regulating FZD4 expression.http://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/a200462.pdfcopd|aec|mir-199a-5p|fzd4|cell apoptosis |
| spellingShingle | WANG Wei, ZHAO Si-lin, FAN Fu-yuan, JIN Zhao-hui, LI Da, FU Yan Down-regulation of miR-199a-5p inhibits lipopolysaccharide- induced apoptosis of rat type Ⅱ alveolar epithelial cell line AECⅡ Jichu yixue yu linchuang copd|aec|mir-199a-5p|fzd4|cell apoptosis |
| title | Down-regulation of miR-199a-5p inhibits lipopolysaccharide- induced apoptosis of rat type Ⅱ alveolar epithelial cell line AECⅡ |
| title_full | Down-regulation of miR-199a-5p inhibits lipopolysaccharide- induced apoptosis of rat type Ⅱ alveolar epithelial cell line AECⅡ |
| title_fullStr | Down-regulation of miR-199a-5p inhibits lipopolysaccharide- induced apoptosis of rat type Ⅱ alveolar epithelial cell line AECⅡ |
| title_full_unstemmed | Down-regulation of miR-199a-5p inhibits lipopolysaccharide- induced apoptosis of rat type Ⅱ alveolar epithelial cell line AECⅡ |
| title_short | Down-regulation of miR-199a-5p inhibits lipopolysaccharide- induced apoptosis of rat type Ⅱ alveolar epithelial cell line AECⅡ |
| title_sort | down regulation of mir 199a 5p inhibits lipopolysaccharide induced apoptosis of rat type ii alveolar epithelial cell line aecii |
| topic | copd|aec|mir-199a-5p|fzd4|cell apoptosis |
| url | http://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/a200462.pdf |
| work_keys_str_mv | AT wangweizhaosilinfanfuyuanjinzhaohuilidafuyan downregulationofmir199a5pinhibitslipopolysaccharideinducedapoptosisofrattypeiialveolarepithelialcelllineaecii |