The saponin D39 blocks dissociation of non‐muscular myosin heavy chain IIA from TNF receptor 2, suppressing tissue factor expression and venous thrombosis

Zhai, Kefeng; Zheng, Jinrong; Ying, Tang; Li, Fang; Lv, Yanni; Zhang, Yuanyuan; Gao, Zhenli; Qi, Jin; Yu, Boyang; Kou, Junping

doi:10.1111/bph.13885

Cited by 59 publications

(39 citation statements)

References 54 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Protein samples were prepared using PRO‐PREP protein extraction solution (Beyotime Institute of Biotechnology) for total fractions according to the manufacturer's instructions 12 . Total protein was separated by sodium dodecyl sulphate‐polyacrylamide gel electrophoresis (SDS‐PAGE) and transferred to polyvinylidene fluoride membranes (Millipore).…”

Section: Methodsmentioning

confidence: 99%

Role of CARD9 in inflammatory signal pathway of peritoneal macrophages in severe acute pancreatitis

Wang

Tian

et al. 2020

J Cellular Molecular Medi

View full text Add to dashboard Cite

Previous studies revealed that caspase recruitment domain protein 9 (CARD9) was involved in severe acute pancreatitis (SAP) inflammation and that interfering with its expression in vivo could inhibit inflammation. However, the specific mechanism is unknown. This study aimed to discover the related signal pathways of CARD9 in macrophages. SiRNA interference technology was used in vivo and in vitro to detect CARD9‐related signal pathways in peritoneal macrophages. Furthermore, Toll‐like receptor 4 (TLR4) and membrane‐associated C‐type lectin‐1 (Dectin‐1) pathways in macrophages were activated specially to looking for the upstream signal path of CARD9. Results showed up‐regulation of CARD9 expression in peritoneal macrophages of SAP rats (P < .05). CARD9 siRNA alleviated inflammatory cytokines, and inhibited the phosphorylation of NF‐κB and p38MAPK in peritoneal macrophages in vivo or in vitro. Meanwhile, CARD9 siRNA reduced the concentration of CARD9 and Bcl10 in peritoneal macrophages, and TLR4 and Dectin‐1 took part in CARD9 signal pathways in macrophages. In conclusion, there is an inflammation signal pathway comprised of TLR4/Dectin‐1‐CARD9‐NF‐κB/p38MAPK activated in macrophages in SAP. Blockade of CARD9 expression in macrophages can effectively alleviate SAP inflammation.

show abstract

Section: Methodsmentioning

confidence: 99%

Role of CARD9 in inflammatory signal pathway of peritoneal macrophages in severe acute pancreatitis

Wang

Tian

et al. 2020

J Cellular Molecular Medi

View full text Add to dashboard Cite

show abstract

“…Cell transfection. Cell transfection was performed according to a previously reported experimental method (32). The miR-15a-5p inhibitor, miR-15a-5p mimic and their respective negative controls (NCs) were purchased from Shanghai GenePharma Co., Ltd.…”

Section: Methodsmentioning

confidence: 99%

Knockdown of lncRNA ANRIL suppresses the production of inflammatory cytokines and mucin 5AC in nasal epithelial cells via the miR-15a-5p/JAK2 axis

Liu

et al. 2020

Mol Med Rep

View full text Add to dashboard Cite

The incidence of allergic rhinitis (AR) is increasing worldwide. Human nasal epithelial cells (HNECs) are the key cells in the occurrence of AR. Antisense non-coding RNA in the INK4 locus (ANRIL) was discovered to be involved in the progression of AR. However, the mechanism by which ANRIL mediates the progression of AR remains to be determined. The present study aimed to further explore the mechanism by which ANRIL regulates AR. Thereby, HNECs were treated with IL-13 to mimic AR in vitro . The mRNA expression levels of ANRIL, microRNA (miR)-15a-5p, JAK2, mucin 5AC (MUC5AC), granulocyte-macrophage colony-stimulating factor (GM-CSF) and eotaxin-1, and protein expression levels of JAK2, STAT3 and phosphorylated-STAT3 in HNECs were analyzed using reverse transcription-quantitative PCR and western blotting, respectively. ELISAs were used to detect the secretory levels of inflammatory cytokines and mucin in cell supernatants. In addition, a dual luciferase reporter assay was used to confirm the downstream target of ANRIL and the target gene of miR-15a-5p. The results revealed that the secretory levels of eotaxin-1, GM-CSF and MUC5AC were significantly upregulated by IL-13 in the supernatant of HNECs. The expression levels of ANRIL and JAK2 were also upregulated in IL-13-induced HNECs, while the expression levels of miR-15a-5p were downregulated. In addition, ANRIL was identified to bind to miR-15a-5p. The IL-13-induced upregulation of eotaxin-1, GM-CSF and MUC5AC mRNA expression and secretory levels was significantly inhibited by the genetic knockdown of ANRIL, while the miR-15a-5p inhibitor effectively reversed this effect. JAK2 was also discovered to be directly targeted by miR-15a-5p. The overexpression of JAK2 significantly suppressed the therapeutic effect of miR-15a-5p mimics on IL-13-induced inflammation in vitro . In conclusion, the findings of the present study suggested that the genetic knockdown of ANRIL may suppress the production of inflammatory cytokines and mucin in IL-13-treated HNECs via regulation of the miR-15a-5p/JAK2 axis. Thus, ANRIL may serve as a novel target for AR treatment.

show abstract

“…293 cells were cultured on 24-well plates to a density of 1×10 6 cells/well and then subjected to gene overexpression as previously reported (22). The recombinant plasmids SLC2A9 pcDNA3.1-T2A-EGFP (Fig.…”

Section: Methodsmentioning

confidence: 99%

“…Immunofluorescence staining was performed according to previously reported protocols (22). Cells were cultured on coverslips and fixed in 4% paraformaldehyde at room temperature for 15 min.…”

Section: Methodsmentioning

confidence: 99%

Urate transport capacity of glucose transporter 9 and urate transporter 1 in cartilage chondrocytes

et al. 2019

View full text Add to dashboard Cite

Chronic gouty arthritis, caused by a persistent increase in, and the deposition of, soluble uric acid (sUA), can induce pathological chondrocyte destruction; however, the effects of urate transport and intracellular sUA on chondrocyte functionality and viability are yet to be fully determined. Thus, the aim of the present study was to investigate the presence and functionality of a urate transport system in chondrocytes. The expression profiles of two primary urate reabsorptive transporters, glucose transporter 9 (GLUT9) and urate transporter 1 (URAT1), in human articular cartilage and chondrocyte cell lines were examined via western blotting, reverse transcription-quantitative PCR, immunohistochemistry and immunofluorescence. Then, chondrocytes were incubated with exogenous sUA at increasing concentrations. Negative control assays were conducted via the specific knockdown of GLUT9 and URAT1 with lentiviral short hairpin (sh)RNAs, and by pretreatment with benzbromarone, a known inhibitor of the two transporters. Intracellular UA concentrations were measured using colorimetric assays. The expression levels of GLUT9 and URAT1 were determined in cartilage tissues and chondrocyte cell lines. Incubation of chondrocytes with sUA led to a concentration-dependent increase in intracellular urate concentrations, which was inhibited by GLUT9 or URAT1 knockdown, or by benzbromarone pretreatment (27.13±2.70, 44.22±2.34 and 58.46±2.32% reduction, respectively). In particular, benzbromarone further decreased the already-reduced intracellular UA concentrations in HC-shGLUT9 and HC-shURAT1 cells by 46.79±2.46 and 39.79±2.22%, respectively. Cells overexpressing GLUT9 and URAT1 were used as the positive cell control, which showed increased intracellular UA concentrations that could be reversed by treatment with benzbromarone. In conclusion, chondrocytes may possess an active UA transport system. GLUT9 and URAT1 functioned synergistically to transport UA into the chondrocyte cytoplasm, which was inhibited by specific gene knockdowns and drug-induced inhibition. These results may be fundamental in the further investigation of the pathological changes to chondrocytes induced by sUA during gouty arthritis, and identified UA transport processes as potential targets for the early control of chronic gouty arthritis.

show abstract

The saponin D39 blocks dissociation of non‐muscular myosin heavy chain IIA from TNF receptor 2, suppressing tissue factor expression and venous thrombosis

Abstract: D39 inhibited tissue factor expression and thrombus formation by modulating the Akt/GSK3β and NF-κB signalling pathways through NMMHC IIA. We identified a new natural product that targeted NMMHC IIA, as a potential treatment for thrombotic disorders and other vasculopathies.

Cited by 59 publications

References 54 publications

Role of CARD9 in inflammatory signal pathway of peritoneal macrophages in severe acute pancreatitis

Role of CARD9 in inflammatory signal pathway of peritoneal macrophages in severe acute pancreatitis

Knockdown of lncRNA ANRIL suppresses the production of inflammatory cytokines and mucin 5AC in nasal epithelial cells via the miR-15a-5p/JAK2 axis

Urate transport capacity of glucose transporter 9 and urate transporter 1 in cartilage chondrocytes

Contact Info

Product

Resources

About