摘要
目的 探讨血小板反应蛋白1(TSP-1)对肾小管上皮细胞(HKCs)表达血管内皮生长因子(VEGF)的影响。方法 实验 分4组:对照组(C组)用无血清培养基(FSM)培养人HKCs;转化生长因子β1(TGF-β1)组(T组)用含TGF-β120ng/ml的FSM培养 HKCs;反义寡核苷酸组(TA组)在T组基础上加TSP- 1反义寡核苷酸;错义寡核苷酸组(TS组)在T组基础上加TSP -1错义寡核苷 酸。上述各组均培养72h后,用免疫细胞化学及RT-PCR方法检测TSP-1和VEGF及它们的mRNA表达的变化。结果 T组与C 组比较,TSP 1表达显著增高(P<0.01),VEGF表达显著下降(P<0.01);TA组阻断TSP-1表达后,与T组比较VEGF表达显著增 加(P<0.01)。结论 TGF-β1能诱导HKCs表达TSP-1、SP-1对VEGF的表达有抑制作用。
Objective To investigate the influence of thrombospondin 1 (TSP-1) on the expression of vascular endothelial growth factor (VEGF) in human renal tubular epithelial cells. Methods Human renal tubular epithelial cell (HKCs) line was cultured in vitro with DMEM/F12 1∶1 medium. HKCs were divided into four groups as follows: normal control group [HKCs were cultured with serum free medium (FSM), C group], TGF-β 1 group (HKCs were treated with FSM containing TGF-β 1, T group), antisense oligonucleotide group [HKCs were treated with FSM contained TGF-β 1 and TSP-1 antisense oligonucleotide (AS), TA group], missense oligonucleotide groups [HKCs were treated with FSM contained TGF-β 1 and TSP-1 missense oligonucleotide (SC), TS group]. Cells were cultured 72 hours in the conditioned media. The expressions of TSP-1 and VEGF were detected with immunohistochemistry. The expressions of TSP-1 mRNA and VEGF mRNA were assayed by RT-PCR. The correlation between the relative expressions of VEGF mRNA and TSP-1 mRNA was analyzed. Results There were significantly increased expressions of VEGF and VEGF mRNA in TA groups (P<0.01) and unchanged expression in TS groups (P>0.05) compared with T groups. The expressions of VEGF and VEGF mRNA were decreased significantly in T groups and TS groups (P<0.01), and were decreased in TA groups (P<0.05) compared with C groups. There was significantly decreased expression of TSP-1 and TSP-1 mRNA in TA groups (P<0.01) but unchanged in TS groups (P>0.05) compared with T groups. The expressions of TSP-1 and TSP-1 mRNA were increased significantly in T groups and TS groups (P<0.01), and they also were increased in TA groups (P<0.05) compared with C groups. Statistical analysis showed that there was a negative correlation between the expression of VEGF mRNA and TSP-1 mRNA (R=-0.898, P<0.01). Conclusion TGF-β 1 induced expression of TSP-1 in HKCs. VEGF expression could be inhibited by TSP-1.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2005年第3期232-236,共5页
Medical Journal of Chinese People's Liberation Army
