The anti-neoplastic activities of aloperine in HeLa cervical cancer cells are associated with inhibition of the IL-6-JAK1-STAT3 feedback loop / 中国天然药物

Cervical cancer (CC) is recognized as the most common neoplasm in the female reproductive system worldwide. The lack of chemotherapeutic agents with outstanding effectiveness and safety severely compromises the anti-cipated prognosis of patients. Aloperine (ALO) is a natural quinolizidine alkaloid with marked anti-cancer effects on multiple malignancies as well as favorable activity in relieving inflammation, allergies and infection. However, its therapeutic efficacy and underlying mechanism in CC are still unclear. In the current study, MTT assay was employed to evaluate the viability of HeLa cells exposed to ALO to preliminarily estimate the effectiveness of ALO in CC. Then, the effects of ALO on the proliferation and apoptosis of HeLa cells were further investigated by plate colony formation and flow cytometry, respectively, while the migration and invasion of ALO-treated HeLa cells were evaluated using Transwell assay. Moreover, nude mice were subcutaneously inoculated with HeLa cells to demonstrate the anti-CC properties of ALO in vivo. The molecular mechanisms underlying these effects of ALO were evaluated by Western blot and immunohistochemical analysis. This study experimentally demonstrated that ALO inhibited the proliferation of HeLa cells via G2 phase cell cycle arrest. Simultaneously, ALO promoted an increase in the percentage of apoptotic HeLa cells by increasing the Bax/Bcl-2 ratio. Additionally, the migration and invasion of HeLa cells were attenuated by ALO treatment, which was considered to result from inhibition of epithelial-to-mesenchymal transition. For molecular mechanisms, the expression and activation of the IL-6-JAK1-STAT3 feedback loop were markedly suppressed by ALO treatment. This study indicated that ALO markedly suppresses the proliferation, migration and invasion and enhances the apoptosis of HeLa cells. In addition, these prominent anti-CC properties of ALO are associated with repression of the IL-6-JAK1-STAT3 feedback loop.

Electroacupuncture protects septic rats from acute lung injury through the JAK1/STAT3 pathway / 南方医科大学学报

OBJECTIVE@#To explore the protective effect of electroacupuncture against acute lung injury (ALI) in septic rats and explore the mechanism.@*METHODS@#Sixty male SD rats were randomly divided into cecal ligation and puncture (CLP)-induced sepsis group (@*RESULTS@#Compared with those in the sham operation group, the rats in ALI group showed obvious lung pathologies with significantly increased lung W/D ratio (@*CONCLUSIONS@#Electroacupuncture can inhibit the release of inflammatory mediators and cell apoptosis via the JAK1/STAT3 pathway to reduce lung injuries in septic rats.

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.

Down-regulation of JAK1/STAT3 signal pathway by miR-34a in PAX6 affects invasion and migration of retinoblastoma / 中国免疫学杂志

Objective:To investigate the effect of miR-34a targeting PAX6 on JAK/STAT signaling pathway on invasion and metastasis of retinoblastoma.Methods:The expression of PAX6 in retinoblastoma tissues was detected by immunohistochemistry.The expression of miR-34a in retinoblastoma cell line was detected by PCR.The effect of miR-34a on the expression of PAX6 was examined by the dual luciferase gene system.Transwell invasion assay and scratch test was used to detect the ability of invasion and migration in the retinoblastoma cell line Rb44 after overexpression miR-34a.Western blot was used to detect the protein expression of JAK/STAT signal pathway after overexpression miR-34a.Results:The expression of PAX6 was significantly higher in retinoblastoma tissues than that in normal tissues.The miR-34a was lower in retinoblastoma tissues than that in normal tissues.Western blot analysis showed the lowest level of PAX6 in Rb44 retinoblastoma.The dual luciferase reporter gene system showed that miR-34a could directly regulate the transcriptional activity of PAX6.The ability of invasion and migration was inhibited after overexpression miR-34a.The expression level of PAX6 was down-regulated and the expression of JAK1/STAT3 protein were down-regulated after overexpression the miR-34a.Conclusion:miR-34a targets the expression of PAX6 and regulates the invasion and migration of retinoblastoma cells by JAK1/STAT3 signal pathway.

Study on the effect of small molecule inhibitors LDYS-14007 on JAK1-STAT3 signaling pathway / 中国生化药物杂志

Objective To study the effect of LDYS-14007 on JAK1-STAT3 signaling pathways.Methods MDA-MB-231 cells were treated with 10μmol,1 nmol LDYS-14007, and 10 μmol Tofacitinib,respectively.Western blot assay was used to determine the expression of JAK1,Phospho-JAK1, STAT3 and Phospho-STAT3.Results The absorbance value was linearly related to the concentration of protein C, The linear equation is A=0.0075C+0.0029, r=0.9976, The linear range of 1.08-5.08 mg/mL, With the increased concentration of LDYS-14007, the amount of Phospho-JAK1, Phospho-STAT3 were all gradually decreased.Conclusion LDYS-14007 leads to the levels of Phospho-JAK1 and Phospho-STAT3 decrease, which inhibits JAK1-STAT3 signaling pathway.LDYS-14007 may play an important role in the treatment of rheumatoid arthritis.

Jak1/Stat3 Is an Upstream Signaling of NF-kappaB Activation in Helicobacter pylori-Induced IL-8 Production in Gastric Epithelial AGS Cells

Helicobacter pylori (H. pylori) induces the activation of nuclear factor-kB (NF-kappaB) and cytokine expression in gastric epithelial cells. The Janus kinase/signal transducers and activators of transcription (Jak/Stat) cascade is the inflammatory signaling in various cells. The purpose of the present study is to determine whether H. pylori-induced activation of NF-kappaB and the expression of interleukin-8 (IL-8) are mediated by the activation of Jak1/Stat3 in gastric epithelial (AGS) cells. Thus, gastric epithelial AGS cells were infected with H. pylori in Korean isolates (HP99) at bacterium/cell ratio of 300:1, and the level of IL-8 in the medium was determined by enzyme-linked immonosorbent assay. Phospho-specific and total forms of Jak1/Stat3 and IkappaBalpha were assessed by Western blot analysis, and NF-kappaB activation was determined by electrophoretic mobility shift assay. The results showed that H. pylori induced the activation of Jak1/Stat3 and IL-8 production, which was inhibited by a Jak/Stat3 specific inhibitor AG490 in AGS cells in a dose-dependent manner. H. pylori-induced activation of NF-kappaB, determined by phosphorylation of IkappaBalpha and NF-kappaB-DNA binding activity, were inhibited by AG490. In conclusion, Jak1/Stat3 activation may mediate the activation of NF-kappaB and the expression of IL-8 in H. pylori-infected AGS cells. Inhibition of Jak1/Stat3 may be beneficial for the treatment of H. pylori-induced gastric inflammation, since the activation of NF-kappaB is inhibited and inflammatory cytokine expression is suppressed.

Jak1/Stat3 Is an Upstream Signaling of NF-kappaB Activation in Helicobacter pylori-Induced IL-8 Production in Gastric Epithelial AGS Cells

Helicobacter pylori (H. pylori) induces the activation of nuclear factor-kB (NF-kappaB) and cytokine expression in gastric epithelial cells. The Janus kinase/signal transducers and activators of transcription (Jak/Stat) cascade is the inflammatory signaling in various cells. The purpose of the present study is to determine whether H. pylori-induced activation of NF-kappaB and the expression of interleukin-8 (IL-8) are mediated by the activation of Jak1/Stat3 in gastric epithelial (AGS) cells. Thus, gastric epithelial AGS cells were infected with H. pylori in Korean isolates (HP99) at bacterium/cell ratio of 300:1, and the level of IL-8 in the medium was determined by enzyme-linked immonosorbent assay. Phospho-specific and total forms of Jak1/Stat3 and IkappaBalpha were assessed by Western blot analysis, and NF-kappaB activation was determined by electrophoretic mobility shift assay. The results showed that H. pylori induced the activation of Jak1/Stat3 and IL-8 production, which was inhibited by a Jak/Stat3 specific inhibitor AG490 in AGS cells in a dose-dependent manner. H. pylori-induced activation of NF-kappaB, determined by phosphorylation of IkappaBalpha and NF-kappaB-DNA binding activity, were inhibited by AG490. In conclusion, Jak1/Stat3 activation may mediate the activation of NF-kappaB and the expression of IL-8 in H. pylori-infected AGS cells. Inhibition of Jak1/Stat3 may be beneficial for the treatment of H. pylori-induced gastric inflammation, since the activation of NF-kappaB is inhibited and inflammatory cytokine expression is suppressed.

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.