Title: Inhibiting effect of adiponectin on high glucose-induced mouse mesangial cells proliferation via microRNA221/tissue inhibitor of metalloproteinase 3 signaling regulation
Abstract: Objective
To investigate the inhibiting effect and mechanism of adiponectin on high glucose - induced mouse mesangial cells proliferation through microRNA221(miR-221)/tissue inhibitor of metalloproteinase 3(Timp3) pathway.
Methods
Mouse mesangial cells cultured in vitro were divided into the following groups: normal control group, high glucose group, high glucose + adiponectin group, high glucose+adiponectin+miR-221 inhibitor transfected group and control group, small interfering RNA-Timp3 (Si-Timp3) transfected group and control group, miR-221 mimic transfected group and control group, high glucose + adiponectin + miR - 221 mimic transfected group and control group. Mouse mesangial cells proliferation was measured by MTT assay. The level of miR-221 and Timp3 mRNA expressions were measured by quantitative real-time polymerase chain reaction (qRT-PCR); Western blotting was performed to detect the expression of Timp3 protein. Dual luciferase reporter assay was used to confirm the activity of 3'-UTR in Timp3. Differences between two groups were compared by LSD-t test.
Results
High glucose could significantly promote the proliferation of mouse mesangial cells, up - regulate the expression of miR-221 and inhibit Timp3 (3.20±0.28vs1, 0.67±0.07vs1, t=13.69, 8.35, all P<0.05). Adiponectin could reverse the effect (t=7.60, 4.12, all P<0.05) . Compared to high glucose+ adiponectin+ miR-221 inhibitor transfected control group, miR-221 inhibitor up-regulated the mRNA expression of Timp3 (0.78 ± 0.03 vs 0.38 ± 0.04, t=14.57, P<0.05) . The cell proliferation was increased significantly in Si-Timp3 transfected group (1.72±0.06 vs 1.04±0.12, t=8.76, P<0.05) . The mRNA level of Timp3 was down-regulated in high glucose+adiponectin+miR-221 mimic transfected group compared to control group (0.72±0.09 vs 1.40±0.06, t=10.93, P<0.05) .
Conclusions
The findings suggest that high glucose is the important factor to improve mouse mesangial cells growth, up-regulate miR-221 but down-regulate Timp3. Adiponectin can reverse the effect through miR-221 and Timp3 pathway by negatively regulates Timp3 expression in high glucose-induced mouse mesangial cells.
Key words:
Adiponectin; microRNA221; Tissue inhibitor of metalloproteinase 3; High glucose; Mesangial cells
Publication Year: 2016
Publication Date: 2016-11-27
Language: en
Type: article
Access and Citation
AI Researcher Chatbot
Get quick answers to your questions about the article from our AI researcher chatbot