Institute of Hematology, the First Hospital Affiliated to Zhejiang University of Chinese Medicine, Hangzhou 310006, China.
Chin J Integr Med. 2012 Aug;18(8):616-20. doi: 10.1007/s11655-012-1179-7. Epub 2012 Aug 2.
To investigate the effects of Panax notoginseng saponins (PNS) on the proliferation and differentiation in NIH3T3 cells.
NIH3T3 cells were treated by various concentrations of PNS 0, 0.05, 0.10, 0.20, and 0.40 g/L. The vitality and proliferation potential of cells were detected by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay, the alkaline phosphatase (ALP) activity was measured by p-nitrophenyl phosphate (pNPP) assay, and the mineralization formation ability was tested for the cellular differentiation toward osteoblast, as well as the expression level of phosphorylated extracellular signal-regulated kinase1/2(P-ERK1/2), extracellular signal-regulated kinase1/2 (ERK1/2) protein kinase was analyzed by Western blot with total cell lysate of NIH3T3 cells treated by PNS.
Both MTT and pNPP assay showed that optical density (OD) values were increased in response to PNS treatment at a dose-dependent pattern. The mineralization formation ability was enhanced in PNS-treated NIH3T3 cells compared with untreated cells. Meanwhile, the expression level of P-ERK1/2 protein kinase was up-regulated in PNS-treated NIH3T3 cells, while, the expression level of ERK1/2 protein kinase revealed no obvious difference with or without PNS treated cells.
PNS could pay a role to promote the proliferation and differentiation in NIH3T3 cells by means of up-regulation of P-ERK1/2 protein kinase.
研究三七总皂苷(PNS)对 NIH3T3 细胞增殖和分化的影响。
用不同浓度的 PNS(0、0.05、0.10、0.20 和 0.40 g/L)处理 NIH3T3 细胞。用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)法检测细胞活力和增殖潜能,用对硝基苯磷酸(pNPP)法测定碱性磷酸酶(ALP)活性,检测细胞向成骨细胞分化的矿化形成能力,以及用 PNS 处理的 NIH3T3 细胞总细胞裂解物进行 Western blot 分析磷酸化细胞外信号调节激酶 1/2(P-ERK1/2)、细胞外信号调节激酶 1/2(ERK1/2)蛋白激酶的表达水平。
MTT 和 pNPP 测定均显示 OD 值随 PNS 处理剂量呈依赖性增加。与未处理细胞相比,PNS 处理的 NIH3T3 细胞的矿化形成能力增强。同时,PNS 处理的 NIH3T3 细胞中 P-ERK1/2 蛋白激酶的表达水平上调,而 ERK1/2 蛋白激酶的表达水平在有无 PNS 处理的细胞中没有明显差异。
PNS 通过上调 P-ERK1/2 蛋白激酶促进 NIH3T3 细胞的增殖和分化。