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Experimental Study on the Effect of Metformin on Autophagy in Reducing the Damage of Trophoblastic Cells Induced by High Glucose |
XIANG Meng, et al |
Xi'an Medical College, Shaanxi Xi'an 710021, China |
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Abstract Objective: To investigate the effect of metformin on the trophoblastic cells injury induced by high glucose and its mechanism. Methods: Human chorionic trophoblastic HTR8-SVneo cells were cultured in vitro to establish a high glucose injury model, which was randomly divided into control group, model group, metformin I group, metformin II group and metformin III group. MTT method was used to detect the change of cell survival rate; Apoptosis was detected by Annexin V-FITC/PI method; the expressions of autophagy related gene 7 (ATG7) and Beclin-1 mRNA were detected by real-time fluorescence quantitative PCR (RT-qPCR); and the expressions of ATG7, Beclin-1, microtubule associated protein light chain 3 I/II (LC3I/II) and nuclear porin 62 (p62) were detected by Western blot. Results: The optimal concentration of high glucose induced model was 30 mmol/L. Compared with those in the control group, the HTR8-SVneo cell survival rate and p62 protein expression level in the model group were significantly lower (P<0.05), while the apoptosis rate, LC3II/LC3I value, Beclin-1, ATG7 mRNA and protein expressions were significantly higher (P<0.05), in addition, compared with those in the model group, HTR8-SVneo cell survival rate and p62 protein expression level in metformin I group, metformin II group and metformin III group increased in turn (P<0.05), while the apoptosis rate, LC3II/LC3I value, Beclin-1, ATG7 mRNA and protein expressions decreased in turn (P<0.05), in a dose-dependent manner. Conclusion: Metformin may reduce the damage of human chorionic trophoblastic cells by inhibiting autophagy induced by high glucose.
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