Expression of circPRKDC in serum of patients with new-onset type 2 diabetes mellitus and its protective effect on palmitate-induced islet cell injury
10.19405/j.cnki.issn1000-1492.2023.10.024
- Author:
Zheng Liu
1
;
Qiyuan Cui
2
;
Yun Zheng
2
;
Mingqing He
2
Author Information
1. Dept of Endocrinology，The First Affiliated Hospital of Soochow University，Suzhou 215006
2. Dept of Gerontology，The First Affiliated Hospital of Soochow University，Suzhou 215006
- Publication Type:Journal Article
- Keywords:
circular RNA;
high fat;
microRNA;
diabetes;
apoptosis
- From:
Acta Universitatis Medicinalis Anhui
2023;58(10):1750-1755
- CountryChina
- Language:Chinese
-
Abstract:
Objective : To investigate the expression of circular RNA ( circRNA) circPRKDC in serum of patients with new-onset type 2 diabetes and its protective effect on palmitic acid-induced islet cell injury and its mechanism.
Methods :Fifty patients with new-onset type 2 diabetes and 50 healthy individuals who underwent physical examination during the same period were selected，and the expression of circPRKDC in serum was detected by real-time quantitative polymerase chain reaction ( qRT-PCR) ．The mouse pancreatic β cell line MIN6 was treated with 110 μmol / L palmitic acid for 24 h to establish cell injury model．Negative control lentivirus or circPRKDC overexpression lentivirus were transfected，namely control group and circPRKDC group，respectively．Flow cytometry was used to determine the apoptosis and intracellular reactive oxygen species ( ROS) levels of cells in each group．Bioinformatics software predicted miRNA with binding sites for circPRKDC．The dual-luciferase reporter gene experiment verified the direct binding of circPRKDC to downstream miRNA．The expression of miRNA in each group of cells was detected by qRT-PCR. Western blot was used to detect the expressions of apoptosis-related factors (Bax，Bcl-2， caspase 8 ) and inflammation-related proteins ( NF-κB，p65 ) in each group of cells.
Results : Compared with healthy subjects，the expression of circPRKDC in serum of patients with type 2 diabetes was significantly lower (P < 0. 01) ．Compared with the control group，the expression of circPRKDC in the circPRKDC group increased (P < 0. 01) ，the level of apoptosis decreased (P＜0. 01) ，and the content of intracellular ROS decreased (P ＜0. 01) . circPRKDC directly bound miR-375 (P ＜0. 01 ) ．Compared with the control group，the expression of miR-375 in cells in the circPRKDC group decreased (P＜0. 01) ，the protein expression of Bcl-2 was up-regulated (P＜0. 01) ， and the protein expressions of Bax，caspase 8，NF-κB and p65 were down-regulated (all P＜0. 01) ．
Conclusion:The expression of circPRKDC is down-regulated in the serum of patients with new-onset type 2 diabetes，and it reduces palmitic acid-induced oxidative stress in islet cells by targeting miR-375，thereby inhibiting apoptosis and in- flammatory responses.
- Full text:2024071219224646578circPRKDC在新发2...诱导胰岛细胞损伤的保护作用_刘铮.pdf
