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Probing the Effect of Syringin on High Glucose-Induced MPC5 Damage in Mouse Podocyte Based on the mTOR/p70S6K Signaling Pathway |
YAO Chunhong, LI Guohong, ZHOU Bo, et al |
Dongxihu District People's Hospital, Hubei Wuhan 430040, China |
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Abstract Objective: To investigate the effects of syringin on the injury of mouse podocyte MPC5 induced by high glucose and mammalian target of rapamycin (mTOR)/p70 ribosomal protein S6 kinase (p70S6K) signal pathway. Methods: MPC5 of mouse podocytes were divided into control group (5.5 mmol/L glucose), high glucose induced group (30mmol/L glucose), syringin low concentration group (2.5μmol/L syringin+30mmol/L glucose), syringin medium concentration group (5μmol/L syringin+30mmol/L glucose), syringin high concentration group (10μmol/L syringin+30mmol/L glucose). Each group added the corresponding reagent or drug of the above concentration to RPMI 1640 medium and cultured for 72h. The absorbance of MPC5 cells in each group was measured by microplate reader, and the cell viability was calculated; the apoptosis of MPC5 cells in each group was determined by flow cytometry; the cytoskeleton morphology of MPC5 cells in each group was observed by phalloidin staining; the expression of mTOR, p70S6K messenger RNA (mRNA) and protein in MPC5 cells of each group were determined by fluorescence quantitative PCR and Western blot. Results: The cytoskeleton morphology of MPC5 cells in the control group was normal, the long axis of the cells was linearly distributed, and the fluorescence staining was obvious; compared with the control group, the cytoskeleton of MPC5 cells in the high glucose induced group was broken, the cells were atrophied, the fluorescence staining was dim, the absorbance value and cell viability were significantly decreased, and the apoptosis rate, mTOR, p70S6K mRNA and protein expression levels were significantly increased (P<0.05); compared with the high glucose induced group, the cytoskeleton structure of MPC5 cells in the syringin low, medium and high concentration groups recovered clearly, the fluorescence intensity gradually became bright, the peduncle tended to be obvious, the absorbance value and cell viability increased in turn, and the apoptosis rate, mTOR, p70S6K mRNA and protein expression levels were decreased in turn (P<0.05). Conclusion: Syringin has a protective effect on MPC5 cells damage induced by high glucose, which can significantly promote MPC5 cells proliferation and inhibit its apoptosis. Its mechanism may be related to the inhibition of syringin on the activation of MPC5 cells mTORC1/p70S6K pathway under high glucose state.
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