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目的:探讨葛根异黄酮对软骨细胞中p38、JNK和ERK信号蛋白表达的影响。方法:采用酶消化原代细胞培养法建立人软骨细胞系,同时检测软骨细胞增殖情况。取第3代对数生长期软骨细胞,制备成单细胞悬液,以1×107孔的细胞密度接种于96孔板中。将细胞分为4组,分别为对照组、10 ng/mL组、20 ng/mL组、40 ng/mL组,每组设4个复孔。对照组加DMEM培养基,10 ng/mL组、20 ng/mL组、40 ng/mL组分别加入含有10 ng/mL、20 ng/mL、40 ng/mL葛根异黄酮的DMEM培养基。采用Western Blotting法检测各组软骨细胞中p38、JNK、ERK信号蛋白的表达情况。结果:加入葛根异黄酮培养液后,20 ng/mL浓度的培养液可促进软骨细胞的增殖,40 ng/mL浓度的培养液开始抑制细胞的生长。与对照组比较,20 ng/mL组p38和JNK蛋白的表达显著降低,40 ng/mL组p38蛋白的表达显著降低,差异均有统计学意义(P<0.05)。结论:葛根异黄酮可以促进软骨细胞的增殖;通过降低MAPKs信号通路中p38、JNK蛋白的表达从而发挥抗软骨退变的作用。
Objective: To investigate the effect of isoprenone on the expression of p38, JNK and ERK in chondrocytes. Methods: Human chondrocytes were established by enzymatic digestion of primary cell culture, and the proliferation of chondrocytes was also detected. The third generation logarithmic growth phase chondrocytes were prepared into single cell suspension and seeded into 96-well plates at a density of 1 × 107 cells. The cells were divided into 4 groups: control group, 10 ng / mL group, 20 ng / mL group and 40 ng / mL group, with 4 replicate wells in each group. The DMEM medium supplemented with 10 ng / mL, 20 ng / mL, 40 ng / mL and 10 ng / mL, 20 ng / mL and 40 ng / mL DMEM was added to the control group. Western blotting was used to detect the expression of p38, JNK and ERK in each group of chondrocytes. Results: After adding kudzu isoflavone medium, the medium of 20 ng / mL could promote the proliferation of chondrocytes, while the medium of 40 ng / mL could inhibit the growth of chondrocytes. Compared with the control group, the expression of p38 and JNK protein in 20 ng / mL group was significantly decreased, while the expression of p38 protein in 40 ng / mL group was significantly decreased (P <0.05). Conclusion: Pueraria isoflavones can promote the proliferation of chondrocytes, and play an anti-cartilage degeneration role by decreasing the expression of p38 and JNK in MAPKs signaling pathway.