论文部分内容阅读
目的观察含穿山龙总皂苷血清对大鼠滑膜细胞株RSC-364血管内皮细胞生长因子(VEGF)mRNA和激活蛋白-1(AP-1)活性的影响,探讨其抑制血管新生的作用机制。方法制备含穿山龙总皂苷和雷公藤(阳性对照)血清。将培养好的大鼠滑膜细胞株RSC-364分为空白对照组、模型对照组、含雷公藤多苷血清组和含穿山龙总皂苷血清组,培养1 h,除空白对照组外,其余各组加入IL-17和TNF-α(均为10μg·L-1)共同孵育24 h。采用实时荧光定量PCR方法检测各组细胞VEGF mRNA表达,凝胶电泳迁移率实验法(EMSA)检测各组细胞核蛋白提取物AP-1的DNA结合活性。结果与空白对照组比较,模型对照组VEGF mRNA表达水平及AP-1 DNA结合活性显著增高(P<0.05,P<0.01);与模型对照组比较,含雷公藤多苷血清组、含穿山龙总皂苷血清组VEGF mRNA表达水平及AP-1 DNA结合活性均降低(P<0.05),且两组间比较,差异无统计学意义。结论含穿山龙总皂苷血清可以抑制VEGF mRNA表达水平及AP-1 DNA结合活性,其机制可能是通过抑制转录因子AP-1来调控血管新生关键因子VEGF产生,进而抑制血管新生。
Objective To investigate the effects of total saponins from Pomace of Salvia miltiorrhiza on the expression of vascular endothelial growth factor (VEGF) mRNA and activator protein-1 (AP-1) in rat synovial cell line RSC-364 and to explore its mechanism of action in inhibiting angiogenesis. Methods The preparation of serum containing total saponins of Tripterygium and Tripterygium wilfordii (positive control). The cultured rat synovial cell line RSC-364 was divided into blank control group, model control group, tripterygium glycosides serum group and Salvia total saponin serum group, cultured for 1 h, except for the blank control group, the rest Group were added IL-17 and TNF-α (both 10μg · L-1) co-incubated for 24 h. Real-time fluorescence quantitative PCR was used to detect the expression of VEGF mRNA in each group. The DNA binding activity of AP-1 was detected by electrophoretic mobility shift assay (EMSA). Results Compared with the blank control group, the expression of VEGF mRNA and the DNA-binding activity of AP-1 in the model control group were significantly increased (P <0.05, P <0.01). Compared with the model control group, the serum levels of Tripterygium glycosides, Saponin serum VEGF mRNA expression and AP-1 DNA binding activity were reduced (P <0.05), and the difference between the two groups was not statistically significant. Conclusions Serum total antisense polysaccharide can inhibit the expression of VEGF mRNA and the DNA binding activity of AP-1. The mechanism may be that the inhibition of transcription factor AP-1 regulates the production of VEGF, a key factor of angiogenesis, and then inhibits angiogenesis.