含药血清对IL-17和TNF-α诱导的大鼠滑膜细胞株RSC-364中NF-κB p65、STAT3及VEGF mRNA表达的影响

[Effects of dioscornin tablet containing serum on NF-kappaB p65, STAT3, and VEGF mRNA expressions in rats' synovial cell strain RSC-364 induced by IL-17 and TNF-alpha].

作者信息

Guo Ya-Chun, Gao Ya-Xian, Song Hong-Ru

机构信息

Department of Pathogen Biology, Chengde Medical College, Hebei 067000, China.

出版信息

Zhongguo Zhong Xi Yi Jie He Za Zhi. 2013 Jun;33(6):814-8.

PMID:23980365

Abstract

OBJECTIVE

To observe the effects of Dioscornin Tablet (DT) containing serum on nuclear factor of kappa B (NF-kappaB) p65, signal transducer and activator of transcription 3 (STAT3), and vascular endothelial growth factor (VEGF) mRNA expressions in rats' synovial cell strain 364 (RSC-364) induced by interleukin-17 (IL-17) and tumor necrosis factor-alpha (TNF-alpha), and to investigate the underlying mechanisms for DT to inhibit angiogenesis of rheumatoid arthritis (RA).

METHODS

In this experiment, the vehicle control group, the cell model group, the DT containing serum group, and the positive control group (Tripterygium containing serum) were set up. The DT containing serum and the Tripterygium containing serum were prepared. The RA cell model was established by IL-17 combined TNF-alpha induced injury in RSC-364. The RA cells were intervened by DT containing serum and Tripterygium containing serum respectively. The DNA binding activity of NF-kappaB p65 was detected using TransAM NF-kappaB p65. The expression of STAT3 was observed using Western blot. The VEGF mRNA expressions were detected by real-time quantitative PCR.

RESULTS

Compared with the vehicle control group, the NF-kappaB p65 activity, the expressions of STAT3 and VEGF mRNA increased significantly in RSC-364 induced by IL-17 +TNF-alpha (P < 0.01, P < 0.05). Compared with the model group, the NF-kappaB p65 activity, the expressions of STAT3 and VEGF mRNA decreased significantly in the DT containing serum group and the positive control group (P < 0.01, P < 0.05). There was no statistical difference between the two groups (P > 0.05).

CONCLUSION

DT inhibited the VEGF mRNA expression through inhibiting the NF-kappaB p65 activity and the STAT3 protein expression in the Janus kinase (JAK)-signal transducer and activating transcription factor pathway, thus inhibiting the angiogenesis of RA.

摘要

目的

观察含薯蓣皂苷片血清对白细胞介素 -17（IL -17）和肿瘤坏死因子 -α（TNF -α）诱导的大鼠滑膜细胞株364（RSC -364）中核因子κB（NF -κB）p65、信号转导子和转录激活子3（STAT3）以及血管内皮生长因子（VEGF）mRNA表达的影响，探讨薯蓣皂苷片抑制类风湿关节炎（RA）血管生成的潜在机制。

方法

本实验设空白对照组、细胞模型组、含薯蓣皂苷片血清组和阳性对照组（含雷公藤多苷血清组）。制备含薯蓣皂苷片血清和含雷公藤多苷血清。采用IL -17联合TNF -α诱导RSC -364损伤建立RA细胞模型。分别用含薯蓣皂苷片血清和含雷公藤多苷血清干预RA细胞。采用TransAM NF -κB p65检测NF -κB p65的DNA结合活性。采用蛋白质免疫印迹法观察STAT3的表达。采用实时定量PCR检测VEGF mRNA表达。

结果

与空白对照组比较，IL -17 +TNF -α诱导的RSC -364中NF -κB p65活性、STAT3及VEGF mRNA表达显著升高（P < 0.01，P < 0.05）。与模型组比较，含薯蓣皂苷片血清组和阳性对照组NF -κB p65活性、STAT3及VEGF mRNA表达显著降低（P < 0.01，P < 0.05）。两组间比较差异无统计学意义（P > 0.05）。