青藤碱对人pbmc增殖的抑制作用.ppt

* * * * * * * 正清风痛宁产品的整体优势与先进性 * 药效成分清楚 质量均一性好 疗效确切 毒副作用小 谢谢 * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * PGE2 (% of control) 0 0 0.1 0.1 1 1 10 10 20 20 40 40 60 60 80 80 100 100 Sinomenine( mM) Sinomenine( mM) LTC4 (% of control) 0 20 0.01 0.1 1 40 60 80 100 Nitric oxide production ( % of control) Sinomenine (mM) * 对致炎性细胞因子合成和基因表达的影响 人外周血单核细胞 抑制IL－1?, TNF-?, IL-6 半抑制浓度在0.5mM出现 ▓ : IL-1?， ● : TNF? ▲ : IL-6 Sinomenine concentrations(mM) Cytokines concentration(% of control) 青藤碱对人体外周血单核细胞的细胞因子的合成 青藤碱对巨噬细胞系统的影响 0 500 1000 1500 2000 2500 3000 3500 LPS L+Sin1 TNF-? concentration (pg/ml) TNF-? Inhibitory rate: Sin1(1mM):93.46%; Sin2(0.5mM)49.46%; Sin3(62.5?M)29.01%. LPS: 5?g/ml Incubation time: 24h 93% 29% 49% L+Sin2 L+Sin3 * 抑制巨噬细胞的环氧化酶生成 Fig.25 Effect of sinomenine on COX-1 and COX-2 enzymatic activity in vitro ▲: sinomenie ▓: indomethacin COX-2 COX-1 * 青藤碱抑制滑膜细胞的增殖 抗增殖 青藤碱通过诱导Hs701.T细胞发挥对IL-1β的抗增殖作用。细胞接种于密度为2.5×10个细胞/ ml的96 - 孔板中，并补充10％FBS的DMEM培养液，孵育24小时，然后附加在无血清培养基中，培养72小时。然后将细胞共培养于10ng/ml的IL-1β和不同浓度的青藤碱（0.125-1mm）。一个优越条件细胞培养实验，在10um的右美托咪定中。用 3H-胸苷掺入法测定青藤碱的抗增殖作用 * 细胞周期试验 G1: 83.69% G2: 7.93%, S: 8.39% Starvation 72hr+IL-1? 24h G1: 87.80% G2: 9.04% S: 3.16% G1: 64.84% G2: 22.45% S: 12.71% G1: 88.59% G2: 7.37% S: 4.04% G1: 89.59% G2: 8.32% S: 2.09% G1: 89.55% G2: 4.70% S: 5.75% 0 hr Cell cycle progression test of Hs701.T cells for determination of genes expression. Cells were seeded in 6-well plate until confluence and incubated in a serum free DMEM at different time intervals for starvation. The proportion of DNA in G1, S and G2/M phase was determined by flow cytometry. In (A) 1 to 5 represented the cells were subjected to starvation for 0, 24, 48, 72 and 96 hr respectively. In (B), cells were starved for 72 hrs and sequentially incubated with IL-1? for 24 hrs. 48 hr 72 hr 96hr 24 hr 青藤碱可抑制人滑膜细胞的IL－1β的表达，从而调节基因表达，抑制信号转导，细胞的粘附和相关蛋白的基因 磷脂酰肌醇4激酶催化，α多肽 鸟嘌呤核苷酸结合蛋白（G蛋白），α刺激活性多肽1 Ras相关C3肉毒杆菌毒素基板2（Rho家族小G