Calenduloside E inhibits lipopolysaccharide-induced inflammatory response by inhibiting activation of ROS-mediated JAK1-stat3 signaling pathway in RAW264.7 cells / 南方医科大学学报

OBJECTIVE@#To investigate the effect of calenduloside E on lipopolysaccharide (LPS)-induced inflammatory response in RAW264.7 cells and explore the underlying molecular mechanism.@*METHODS@#CCK-8 assay was used to examine the effect of different concentrations of calenduloside E (0-30 μg/mL) on the viability of RAW264.7 cells. The release of the pro-inflammatory cytokines tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in RAW264.7 cells in response to pretreatment with 6, 8, and 10 μg/mL calenduloside E for 2 h followed by stimulation with 100 ng/mL LPS was detected using enzyme-linked immunosorbent assay (ELISA). The expression levels of iNOS and COX-2 and the activation of JAK-stats, MAPKs and NF-кB signaling pathways in the treated cells were determined using Western blotting. A reactive oxygen species (ROS) detection kit was used to detect ROS production in the cells, and the nuclear translocation of the transcription factor stat3 was observed by laser confocal microscopy.@*RESULTS@#Calenduloside E below 20 μg/mL did not significantly affect the viability of RAW264.7 cells. Calenduloside E dose-dependently decreased the expression levels of iNOS and COX-2 induced by LPS, inhibited LPS-induced release of TNF-α and IL-1β, and suppressed LPS-induced JAK1-stat3 signaling pathway activation and stat3 nuclear translocation. Calenduloside E also significantly reduced ROS production induced by LPS in RAW264.7 cells.@*CONCLUSIONS@#Calenduloside E inhibits LPS-induced inflammatory response by blocking ROS-mediated activation of JAK1-stat3 signaling pathway in RAW264.7 cells.

Effect of lncRNA HULC knockdown on rat secreting pituitary adenoma GH3 cells

Pituitary adenoma is one of the most common tumors in the neuroendocrine system. This study investigated the effects of long non-coding RNAs (lncRNAs) highly up-regulated in liver cancer (HULC) on rat secreting pituitary adenoma GH3 cell viability, migration, invasion, apoptosis, and hormone secretion, as well as the underlying potential mechanisms. Cell transfection and qRT-PCR were used to change and measure the expression levels of HULC, miR-130b, and FOXM1. Cell viability, migration, invasion, and apoptosis were assessed using trypan blue staining assay, MTT assay, two-chamber transwell assay, Guava Nexin assay, and western blotting. The concentrations of prolactin (PRL) and growth hormone (GH) in culture supernatant of GH3 cells were assessed using ELISA. The targeting relationship between miR-130b and FOXM1 was verified using dual luciferase activity. Finally, the expression levels of key factors involved in PI3K/AKT/mTOR and JAK1/STAT3 pathways were evaluated using western blotting. We found that HULC was highly expressed in GH3 cells. Overexpression of HULC promoted GH3 cell viability, migration, invasion, PRL and GH secretion, as well as activated PI3K/AKT/mTOR and JAK1/STAT3 pathways. Knockdown of HULC had opposite effects and induced cell apoptosis. HULC negatively regulated the expression of miR-130b, and miR-130b participated in the effects of HULC on GH3 cells. FOXM1 was a target gene of miR-130b, which was involved in the regulation of GH3 cell viability, migration, invasion, and apoptosis, as well as PI3K/AKT/mTOR and JAK1/STAT3 pathways. In conclusion, HULC tumor-promoting roles in secreting pituitary adenoma might be via down-regulating miR-130b, up-regulating FOXM1, and activating PI3K/AKT/mTOR and JAK1/STAT3 pathways.

Protection of salvianolic acid B on isoproterenol-induced cardiomyocyte hypertrophy of neonatal rats / 中草药

Objective: To observe the effects of salvianolic acid B (Sal B) on isoproterenol (ISO)-induced cardiomyocyte hypertrophy of neonatal rats and clarify the underlying mechanisms. Methods: Hypertrophy in neonatal rat ventricular myocytes was induced by ISO. The effect of Sal B on the myocardial viability of neonatal rats was measured by MTT. The mRNA expression levels of ANP and BNP were detected by RT-PCR. Colorimetric method was employed to measure SOD activity and MDA content. The expression levels of JAK1 and STAT3 were assessed by Western blotting. Results: Sal B at different concentration had no effect on the myocardial viability of neonatal rats. Compared with the model group, Sal B at 10 and 20 μmol/L could obviously down-regulate the gene expression levels of ANP and BNP (P < 0.01, 0.05), significantly increase SOD activity, and decrease MDA content. The protein expression levels of JAK1/STAT3 were down-regulated (P < 0.01, 0.05). Conclusion: Sal B could effectively inhibit ISO-induced cardiomyocyte hypertrophy of neonatal rats and the mechanism may be related with the anti-oxidative stress and the inhibition of JAK1/STAT3 signaling pathway.

Effect of hypericin on myocardial fibrosis of chronic viral myocarditis of mice and its mechanism / 中草药

Objective: To investigate the role of JAK/STAT signal pathway in myocardial fibrosis (MF) of the chronic viral myocarditis (VMC) of mice and hypericin intervention study. Methods: Sixty healthy male Balb/c mice were used to establish the MF model by intermittent multiple ip injection of coxsackie B3, another 10 mice were used as normal control. Two months after the modeling, survival mice were randomly divided into four groups, model group, low- or high-dose hypericin group, and Captopril group. The mice were treated by Captopril or hypericin, respectively, ig administration, once a day. After 30 d, we took the myocardium of left ventricle to dye with Masson, to observe the cardiac histological changes. The serum type I collagen and type III collagen were detected by the means of ELISA, and the expression of JAK1 and STAT3 was observed with semi-quantitative RT-PCR and immunohistochemistry technique. Results: The model group, serum type I and type III collagen increased significantly, the expression level of JAK1/STAT3 was higher than that of the normal group, and the difference was significant (P < 0.05). While the hypericin and Captopril treatment could significantly reduce serum expression of type I and, type III collagen, and decrease JAK1/STAT3 expression. Histology showed the improvement of myocardial fibrosis degree, and a significant difference was observed when comparing with the model group (P < 0.05). Conclusion: In the process of chronic viral myocarditis, the activation of JAK1/STAT3 pathway may be one of pathological mechanisms of the MF-induced with type I and type III collagen increasing. Hypericin could inhibit the myocardial fibrosis by blocking JAK1/STAT3 pathway.