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Thyroid papillary carcinoma is the most common type of thyroid malignancy. The mitogen-activated protein kinase signaling cascade is the major pathway involved in thyroid papillary carcinoma and may be regulated by small molecules and transcription factors. Zerumbone is a bioactive compound with multiple pharmacological properties. This study investigated the effects of zerumbone on survival and proliferation of thyroid papillary carcinoma-1 cells with an emphasis on the level of microRNA- 758–3p and its target gene, mitogen-activated protein kinase-1. Zerumbone exhibited dose-dependent inhibitory effects on the viability and proliferation of thyroid papillary carcinoma-1 cells. It also gradually elevated the level of microRNA-758–3p but decreased mitogen-activated protein kinase-1 expression in these cells. Mitogen-activated protein kinase-1 has been suggested as a downstream target of microRNA-758–3p and confirmed by dual-reporter luciferase assay. The downregulation of microRNA-758–3p in thyroid papillary carcinoma-1 cells via the transfection with microRNA-758–3p inhibitor reversed the inhibitory effect of zerumbone on mitogen-activated protein kinase-1 expression. Furthermore, the transfection of thyroid papillary carcinoma-1 cells with lentiviral vector overexpressing mitogen-activated protein kinase-1 eliminated the protective effects of zerumbone on thyroid papillary carcinoma progression as compared to the group delivered with control vector. In conclusion, this study demonstrated that zerumbone inhibited survival and proliferation of thyroid papillary carcinoma-1 cells via suppressing the expression of mitogen-activated protein kinase-1 through microRNA-758–3p. These data suggest the therapeutic potential of zerumbone in thyroid papillary carcinoma treatment. [ABSTRACT FROM AUTHOR]

Objective. Coronary heart disease (CHD) has been regarded as a serious and common disease in the modern society. This study aims to investigate the effect of Granule of BU-XIN RUAN-MAI (BXRM) on angina pectoris of coronary heart disease and to explore the molecular mechanisms underlying Granule of BU-XIN RUAN-MAI-mediated protective activity against this disease. Methods. The effects of Granule of BU-XIN RUAN-MAI on clinical symptoms of patients' angina were indicated by hemorheology indicators including high shear of blood viscosity, low shear of blood viscosity, plasma viscosity, erythrocyte rigidity index, D-D dimer, fibrinogen content, and lipid content. The effects of Granule of BU-XIN RUAN-MAI on isoprenaline-induced myocardial cell injury were determined by conducting H&E staining and by performing ELISA to examine the serum content of MDA, SOD, Na+/K+-ATPase, cAMP, and the content of inflammatory factors in isoprenaline-induced rats. Meanwhile, western blot and real time PCR were used to determine the expression of genes involved in oxidation and energy metabolism, and real time PCR was also used for determination of miR-542-3p expression. Luciferase reporter assay was conducted to test the binding sites of miR-542-3p on GABARAP 3′UTR. The chemical compositions of Granule of BU-XIN RUAN-MAI were determined by liquid LC-QTOF-MS. Results. Granule of BU-XIN RUAN-MAI significantly attenuated the clinical symptoms of patients' angina by improving the patients' heart rate and by decreasing the level of hemorheology indicators and also by reducing the serum content of TC, TG, LDL, and elevated HDL content. H&E staining demonstrated that Granule of BU-XIN RUAN-MAI ameliorated the myocardial ischemia in a dose-dependent manner. Besides, Granule of BU-XIN RUAN-MAI downregulated serum MDA content and upregulated the content of SOD, Na+/K+-ATPase, and cAMP in isoprenaline-induced rats. Granule of BU-XIN RUAN-MAI significantly improved oxidation stress by increasing PPARα expression, and it inhibited inflammation by downregulating expression and contents of IL-6, IL-1β, and TNF-α. Then, Granule of BU-XIN RUAN-MAI-containing serum increased the SOD content, and reduced the MDA content in angiotensin II-stimulated HUVEC cells. The granule of BU-XIN RUAN-MAI-containing serum obviously downregulated protein expressions of P40phox, P47phox, and P67phox in plasma membrane, and it significantly increased protein levels of P40phox, P47phox, and P67phox in the cytoplasm of HUVEC cells. Furthermore, GABARAP was reduced in heart tissues of ISO-induced rats and in angiotensin II-stimulated cell lines, and GABARAP was required for the inhibitory activity of Granule of BU-XIN RUAN-MAI on oxidation and inflammation in vivo and in vivo. GABARAP could be upregulated by Granule of BU-XIN RUAN-MAI by inhibiting the expression of miR-542-3p, which may significantly enhance oxidation and inflammation by targeting GABARAP in cardiomyocytes. Moreover, the silencing of GABARAP could obviously reverse the granule of BU-XIN RUAN-MAI-mediated protective activity against coronary heart disease, and interfering GABARAP expression also could partly block the anti-miR-542-3p-controlled oxidation and inflammation in cardiomyocytes. Besides, salidroside, loganin, and polydatin were the main compounds of granules of BU-XIN RUAN-MAI. Conclusion. Granule of BU-XIN RUAN-MAI is an excellent prescription for treatment of coronary heart disease by suppressing inflammation and NAPDH-mediated oxidative stress. The miR-542-3p/GABARAP axis is required for Granule of BU-XIN RUAN-MAI, exhibiting its protective activity against the pectoris of coronary heart disease. [ABSTRACT FROM AUTHOR]

MicroRNA (miRNA) can regulate the expression of its target gene by mediating mRNA cleavage or by translational repression at a post-transcriptional level. Usually, one miRNA may regulate many genes as its targets, while one gene may also be targeted by many miRNAs. We previously demonstrated that cyb561d2 , whose protein product is involved in cell defense, and chemical stress, is targeted by miR-155 in adult zebrafish ( Danio rerio ) when exposed to fipronil (5-amino-1-[2,6-dichloro-4-(trifluoromethyl) phenyl]-4-[(trifluoromethyl) sulphinyl]-1H-pyrazole-3-carbonitrile). Microcosm Targets prediction showed that the cyb561d2 gene is also highly possibly targeted by miR-194a , miR-216b , miR-429 , and miR-499 . These interactions need to be further validated experimentally. In this study, we evaluated the effects of fipronil on miR-194a , miR-216b , miR-429 , miR-499 and cyb561d2 in zebrafish and investigated whether these four miRNAs c ould regulate the expression of cyb561d2 in both mRNA and protein levels. The expression of cyb561d2 was upregulated in both mRNA and protein level in a dose-dependent manner upon stimulation of fipronil, and miR-216b and miR-499 were downregulated concurrently, whereas there was no significant changes were observed in the expression level of miR-194a and miR-429 . The dual luciferase report assay demonstrated that miR-216 b and miR-499 interacted with cyb561d2 3′-untranslated regions (3′-UTR), miR-194a and miR-429 did not stimulate degradation of cyb561d2 mRNA. The expression of cyb561d2 was reduced in both mRNA and protein level when ZF4 cells were transfected with miR-499 mimic, whereas expression level of both mRNA and protein was increased when endogenous miR-499 was inhibited by transfection with miR-499 inhibitor. Likewise, the mRNA and protein level of cyb561d2 was affected by treatment with the mimics and the inhibitor of miR-216b . In contrast, when ZF4 cells were transfected with a mimic of miR-194a or miR-429 , the expression of cyb561d2 mRNA was not significantly changed. As a result, cyb561d2 is targeted by miR-155 , miR-216b and miR-499 upon fipronil exposure, and miR-194a and miR-429 can not target cyb561d2 . The expression pattern of these 3 miRNAs presents novel fipronil responses that could be used as a toxicological biomarker. [ABSTRACT FROM AUTHOR]