【摘要】 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。更多还原

【Abstract】 Objective: To discuss the effect of kudzu root isoflavones on the expressions of signal protein p38, JNK and ERK in chondrocyte. Methods: Used the method of culturing primary cells with enzyme digestion to establish human chondrocyte cell lines, and detected the chondrocyte proliferation. Chondrocyte of the third passage in logarithmic growth phase were made into single cell suspension and were inoculated in 96-well plates with the density of 1×10~7cell/mL. Divided all the cells into four groups, namely the control group, 10 ng/mL group, 20 ng/mL group, and 40 ng/mL group, each group being set with four parallel holes. The control group was added with DMEM culture media, while 10 ng/mL group, 20 ng/mL group and 40 ng/mL group were added with DMEM culture media containing kudzu root isoflavones of 10 ng/mL, 20 ng/mL and 40ng/mL dosages. Western Blotting method was adopted to determine the expressions of signal protein p38, JNK and ERK in chondrocyte in each group. Results: After the addition of kudzu root isoflavones into culture media, the culture media of 20 ng/mL dosage promoted chondroyte proliferation, while the culture media of 40 ng/mL dosage started to inhibit the chondroyte proliferation. Comparing with that in the control group, the expressions of protein p38 and JNK were significantly reduced in 20 ng/mL group, and expression of protein p38 was obviously declined in 40 ng/mL group, differences being significant(P<0.05). Conclusion: Kudzu root isoflavones can promote chondroyte proliferation. It prevents cartilage degeneration by decreasing the expressions of protein p38 and JNK in MAPKs signal pathway.更多还原