doi: 10.3967/bes2020.044
Dexmedetomidine Attenuates High Glucose-induced HK-2 Epithelial-mesenchymal Transition by Inhibiting AKT and ERK
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Abstract:
Objective To explore the protective effects of dexmedetomidine (Dex) against high glucose-induced epithelial-mesenchymal transition in HK-2 cells and relevant mechanisms. Methods HK-2 cells were exposed to either glucose or glucose+Dex for 6 h. The production of ROS, morphology of HK-2 cells, and cell cycle were detected. Moreover, the expression of AKT, p-AKT, ERK, p-ERK, PI3K, E-Cadherin, Claudin-1, and α-SMA were determined and compared between HK-2 cells exposed to glucose and those exposed to both glucose and Dex with or without PI3K/AKT pathway inhibitor LY294002 and ERK pathway inhibitor U0126. Results Compared with HK-2 cells exposed to high level of glucose, the HK-2 cells exposed to both high level of glucose and Dex showed: (1) lower level of ROS production; (2) cell morphology was complete; (3) more cells in G1 phase; (4) lower expression of p-AKT, p-ERK and α-SMA, higher expression of E-Cadherin and Claudin-1. PI3K/AKT inhibitor LY294002 and ERK inhibitor U0126 decreased the expression of p-AKT, p-ERK and α-SMA, and increased the expression of E-Cadherin and Claudin-1. Conclusion Dex can attenuate high glucose-induced HK-2 epithelial-mesenchymal transition by inhibiting AKT and ERK. -
Key words:
- Dexmedetomidine /
- Epithelial-mesenchymal transition /
- High glucose /
- Oxidative stress /
- HK-2 cells
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Figure 5. Expression of AKT, ERK, and PI3K in HK-2 cells exposed to glucose and Dex
(A) Western blot analysis of p-AKT and AKT expression in HK-2 cells exposed to glucose and Dex. (B) The comparison of AKT relative expression level in HK-2 cells. (C) The comparison of p-AKT relative expression level in HK-2 cells. (D) The comparison of p-AKT/AKT relative expression level in HK-2 cells. (E) Western blot analysis of p-ERK and ERK expression in HK-2 cells. (F) The comparison of ERK relative expression level in HK-2 cells. (G) The comparison of p-ERK relative expression level in HK-2 cells. (H) The comparison of p-ERK/ERK relative expression level in HK-2 cells. (I) Western blot analysis of PI3K expression in HK-2 cells. (J) The comparison of PI3K relative expression level in HK-2 cells. *P < 0.05; **P < 0.01; Data are the mean ± SD, n = 3.
Figure 6. Expression of AKT, ERK, E-Cadherin, Claudin-1, and α-SMA in HK-2 cells exposed to glucose, Dex or PI3K/AKT inhibitor. (A) Western blot analysis of p-AKT, p-ERK, E-Cadherin, α-SMA, and Claudin-1. (B) The comparison of p-Akt/Akt relative expression level in HK-2 cells. (C) The comparison of p-ERK/ERK relative expression level in HK-2 cells. (D) The comparison of E-Cadherin relative expression level in HK-2 cells. (E) The comparison of α-SMA relative expression level in HK-2 cells. (F) The comparison of Claudin-1 relative expression level in HK-2 cells.
*P < 0.05; **P < 0.01; ***P < 0.001; Data are the mean ± SD, n = 3.
Figure 7. Expression of ERK, AKT, E-Cadherin, α-SMA, and Claudin-1 in HK-2 cells exposed to glucose, Dex or ERK inhibitor. (A) Western blot analysis of p-ERK, p-AKT, E-Cadherin, α-SMA, and Claudin-1. (B) The comparison of p-ERK/ERK relative expression level in HK-2 cells. (C) The comparison of p-AKT/AKT relative expression level in HK-2 cells. (D) The comparison of E-Cadherin relative expression level in HK-2 cells. (E) The comparison of α-SMA relative expression level in HK-2 cells. (F) The comparison of Claudin-1 relative expression level in HK-2 cells.
*P < 0.05; **P < 0.01; ***P < 0.001; Data are the mean ± SD, n = 3.
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