Hyperoside Alleviates High Glucose-Induced Proliferation of Mesangial Cells through the Inhibition of the ERK/CREB/miRNA-34a Signaling Pathway

Figure 3

Hyperoside regulates miR-34a through the ERK/CREB pathway. (a) SV40-MES13 cells were treated with LG or HG with different concentrations of hyperoside for 48 h. The phosphorylated CREB was examined using Western blot. (b) SV40-MES13 cells treated with or without U0126 (10 μM) were concurrently treated with or without hyperoside (200 μM) for 48 h under HG condition. The phosphorylation level of CREB was examined by Western blot. (c) SV40-MES13 cells were treated or untreated with hyperoside (200 μM) in the absence or presence of U0126 (10 μM). The qRT-PCR analysis of sheared DNA from control and hyperoside-treated cells after ChIP with antibody directed against CREB. (d–g) Protein expression of CREB and the phosphorylation level of ERK in CREB-shRNA or pcDNA3.1-HA-CREB expression plasmids transfected cells were detected by Western blot, and (h, i) the expression of miR-34a was detected by qRT-PCR. (j, k) The expression of miR-34a in antagomir-34a or agomir-34a transfected cells was detected by qRT-PCR, and (l, m) the phosphorylation levels of ERK and CREB were detected by Western blot (, , , n.s. indicates no significance).

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