2018•Zhonghua shiyong erke linchuang zazhiRequires access
Influence of endothelin-1 on nitric oxide and hydrogen sulfide pathways in vascular smooth muscle cells in rats
Xiao Yu Tian, Qingyou Zhang, Yaqian Huang, Da Zhang, Xinjing Tang, Hongfang Jin, Junbao Du, Chaoshu Tang, Yan Sun
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Abstract
Objective
To explore the effect of different concentrations of endothelin-1 (ET-1) on the endogenous nitric oxide (NO) and hydrogen sulfide (H2S) pathways of vascular smooth muscle cells (A7r5 cell lines)in rats.
Methods
A7r5 cell lines were divided into the control group and the experimental group.ET-1 at a concentration of 10-8 -10-6 mol/L was added into the experimental group, and as for the control group, the same volume of sterile phosphate buffered saline (PBS) buffer solution was added.The content of NO and H2S in A7r5 cell lines was detected by fluorescent NO probe and H2S probe after ET-1 stimulation for 48 h, respectively.The content of NO in the supernatant was measured by NO assay kit at 48 h of the incubation.The content of H2S in the supernatant was measured by polarographic H2S sensor at 48 h of the incubation.The expressions of inducible nitric oxide synthase (NOS2), endothelial nitric oxide synthase (NOS3), cystathionine-γ-lyase (CSE), cystathionine-β-synthase (CBS) and proliferating cell nuclear antigen (PCNA) were detected by the Western blot method.
Results
The relative fluorescence intensity of the content of NO in the A7r5 cell lines of ET-1 10-8, 10-7 and 10-6 mol/L groups (0.078±0.080, 0.075±0.002, 0.056±0.009) was markedly lower than that in the control group(0.094±0.061), and the differences were statistically significant(F=15.248, P 0.05). The relative fluorescence intensity of the content of H2S in the A7r5 cell lines of ET-1 10-8, 10-7 and 10-6 mol/L groups (0.063±0.002, 0.056±0.008, 0.042±0.009) was markedly lower than that in the control group (0.082±0.006), and the differences were statistically significant(F=16.297, P 0.05). The data showed that PCNA protein expression in the 10-7 mol/L ET-1 group(0.686±0.180)significantly increased compared with that of the control group(0.437±0.191), and the difference was statistically significant (t=-2.840, P<0.01).
Conclusion
ET-1 stimulation can lead to the proliferation of vascular smooth muscle cells and down-regulate its endogenous NO and H2S pathways.
Key words:
Endothelin-1; Nitric oxide; Hydrogen sulfide; Vascular smooth muscle cells; Rat