Salidroside inhibits phenotypic transformation of rat pulmonary artery smooth muscle cells induced by hypoxia / 中国中药杂志

This study investigated the effect of salidroside on phenotypic transformation of rat pulmonary artery smooth muscle cells(PASMCs) induced by hypoxia. Rat pulmonary arteries were isolated by tissue digestion and PASMCs were cultured. The OD values of cells treated with salidroside at different concentrations for 48 hours were measured by cell counting kit-8(CCK-8) to determine the appropriate concentration range of salidroside. The cells were divided into a normal(normoxia) group, a model(hypoxia) group, and three hypoxia + salidroside groups(40, 60, and 80 μg·mL~(-1)). Quantitative real-time PCR(qRT-PCR) was used to detect the mRNA expression of cell contractile markers in each group, such as α-smooth muscle actin(α-SMA), smooth muscle 22(SM22), and calcium-binding protein(calponin), and synthetic marker vimentin. The expression levels of cell phenotypic markers and proliferating cell nuclear antigen(PCNA) were detected by Western blot. The proliferation of cells in each group was detected by the 5-ethynyl-2'-deoxyuridine(EdU) assay. Cell migration was measured by Transwell assay. As revealed by results, compared with the normal group, the model group showed decreased mRNA and protein expression of contractile phenotypic markers of PASMCs and increased mRNA and protein expression of synthetic markers. Compared with the conditions in the model group, salidroside could down-regulate the mRNA and protein expression of synthetic markers in PASMCs and up-regulated the mRNA and protein expression of contractile phenotypic markers. Compared with the normal group, the model group showed potentiated proliferation and migration. Compared with the model group, the hypoxia + salidroside groups showed blunted proliferation and migration of cells after phenotypic transformation. The results suggest that salidroside can inhibit the expression of synthetic markers in PASMCs and promote the expression of contractile markers to inhibit the hypoxia-induced phenotypic transformation of PASMCs. The mechanism of salidroside in inhibiting the proliferation and migration of PASMCs is related to the inhibition of the phenotypic transformation of PASMCs.

Effect of miR-34a on Proliferation of Pulmonary Artery Smooth Muscle Cells in Rats Induced by Hypoxia / 中山大学学报(医学科学版)

@#To observe the effect of miR-34a on the proliferation of pulmonary artery smooth muscle cells in rats induced by hypoxia and explore its possible mechanism.【Methods】Rat pulmonary artery smooth muscle cells were primarily isolated from pulmonary arteriole and cultured. After 3% O2 treatment，the expression of miR- 34a and Notch1 mRNA in rat PASMC were detected by real time PCR. The cell proliferation was detected by EDU after over-expression and inhibition of miR-34a and silencing Notch1 by cell transfection under hypoxia，and the expression of PCNA was detected by real time PCR and western blot method.【Results】We successfully isolated and cultured rat PASMC. And after 3% O2 treatment，the expression of miR-34a in rat PASMC was significantly decreased after 48 h compared with 24 h（P < 0.05）. However，the expression of Notch1 mRNA increased significantly after 48 h compared with 24 h（P < 0.05）. In addition， over-expression of miR-34a and silencing Notch1 significantly inhibited hypoxia-induced cell proliferation ，while inhibition of miR-34a significantly promoted the PASMC proliferation（P < 0.05）.【Conclusion】miR-34a participates in the proliferation of PASMC induced by hypoxia，and it may be through up-regulation of Notch1 to induce cell proliferation.