ABSTRACT
Stroke is a multifactorial disease and a consequence of morbidities of diabetes, obesity, hypertension, and heart diseases. Leptin is a major adipokine that regulates weight balance and energy homeostasis, the level of which has been considered as an indicator of acute ischemic stroke. In the present study, we confirmed the high level of leptin and noradrenaline in stroke patients and mouse models as well as oxygen-glucose deprivation (OGD) primary cerebral neurons. Leptin administration increased noradrenaline concentration and dopamine ß-monooxygenase (DBH) but decreased noradrenaline transporter (NET) expression in primary cerebral neurons. Moreover, induced noradrenaline concentration, DBH activity, and inhibited NET were blunted by TG101348 (JAK2 inhibitor). JAK2 silencing also abolished the effects of leptin on noradrenaline metabolism.
Subject(s)
Brain Ischemia/drug therapy , Brain Ischemia/metabolism , Leptin/pharmacology , Norepinephrine/metabolism , Animals , Case-Control Studies , Cells, Cultured , Dopamine beta-Hydroxylase/metabolism , Humans , Janus Kinase 2/antagonists & inhibitors , Janus Kinase 2/metabolism , Male , Mice , Neurons/cytology , Neurons/metabolism , Norepinephrine Plasma Membrane Transport Proteins/metabolism , Pyrrolidines/pharmacology , Sulfonamides/pharmacologyABSTRACT
Objective: To investigate the effects of S100A6 gene silencing on the proliferation and migration of Eca109 human esophagus cancer cells. Methods: The shRNA expression vectors were constructed using the shRNA sequences designed based on human S100A6's coding sequence, and were transfected into Eca109 cells via cationic liposome. The changes of S100A6 mRNA and protein in Eca109 cells transfected with the recombinant vectors were detected using real-time PCR and Western blotting analysis 48 hours after transfection, respectively; the proliferative curves of transfected cells were plotted using MTT assay; furthermore, the change in cellular migration ability was determined using wound healing assay. Results: The eukaryotic expression vector of shRNA targeting S100A6 was successfully constructed. Real-time PCR and Western blotting analysis results showed that S100A6 was effectively silenced by liposome-mediated transfection of the recombinant shRNA vectors in Eca109 cells. Compared with the untransfected cells, S100A6 mRNA and protein in transfected Eca109 cells were significantly decreased (P<0.05, P<0.01). Meanwhile, the proliferative activity of Eca109 cells was significantly inhibited by S100A6 silencing (P<0.01). It was found that the cellular migration was also suppressed by S100A6 gene interference. Conclusion: S100A6 gene can be effectively silenced by shRNA expression vectors, and the silence may lead to inhibition of the proliferation and migration of Eca109 cells.
ABSTRACT
Objective: To investigate the influence of TTS-12, a steroid saponin from Tribulus terrestris L., on the formation of Candida albicans biofilm, and to discuss the possible mechanism. Methods: The inhibition of C. albicans biofilm formation by TTS -12 was observed by confocal scanning laser microscopy; XTT method was used to investigate the influence of different concentrations of TTS-12 on C. albicans biofilm formation. The water-hydrocarbon two-phase assay was used to measure the cell surface hydrophobicity of C. albicans treated with different concentrations of TTS-12. The expression of CSH1 mRNA was measured by real-time RT-PCR. Results: Compared with control group, TTS-12 treatment resulted in loose C. albicans biofilm, and it dose-dependently inhibited C. albicans biofilm formation. The cell surface hydrophobicity in the TTS-12-treated groups was lower than that in the control group; consistent with this, TTS-12-treated cells also expressed significantly lower levels of CSH1 mRNA than cells in control group (P<0. 01). Conclusion: TTS-12 may inhibit the formation of C. albicans biofilm through inhibiting CSHI gene expression.
