牛膝提取物对碘乙酸钠诱导的软骨细胞保护作用  

Protective Effect of Achyranths Bidentatae Radix Extracts on Monosodium Iodoacetate-induced Chondrocytes Injury

作　　者：李淑洁[1] 陈秀娟[1] 任艳红[1] 王伟[2] 

机构地区：[1]莱芜市人民医院,山东莱芜271100 [2]山东中医药大学附属医院,济南250011

出　　处：《中国实验方剂学杂志》2015年第14期132-135,共4页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：山东省自然科学基金项目(Y2006C02)

摘　　要：目的:探讨牛膝提取物对碘乙酸钠(MIA)诱导的软骨细胞保护作用。方法:通过Ⅱ型胶原酶消化SD大鼠软骨,获得软骨细胞,实验分为5组,分别为正常组,模型组(4μmol·L-1MIA),牛膝提取物低剂量组(10 mg·L-1+4μmol·L-1MIA),牛膝提取物中剂量组(30 mg·L-1+4μmol·L-1MIA),牛膝提取物高剂量组(100 mg·L-1+4μmol·L-1MIA),造模与给药分别为24 h;MTT法检测各组软骨细胞活力;Hoechst 33258染色法检测各组软骨细胞形态变化;分光光度法检测各组软骨细胞中Caspase-3活性;Western blot分析各组AKT激活状况及下游靶分子Bax,Bcl-2的表达。结果:与正常组比较,模型组中软骨细胞活力下降,细胞核发生皱缩或裂解,细胞中Caspase-3活性和Bax表达量上升,Bcl-2表达量下降,AKT磷酸化程度降低,差异均具有显著性意义(P<0.01)。与模型组比较,牛膝提取物中、高剂量组可改善软骨细胞活力,恢复软骨细胞形态并抑制Bax表达;牛膝提取物低、中、高剂量组皆能降低大鼠软骨细胞中Caspase-3活性,提高Bcl-2表达,并促进AKT磷酸化,差异具有显著性意义(P<0.01)。结论:牛膝提取物具有保护MIA诱导的软骨细胞作用,是通过抑制软骨细胞凋亡实现的,可能与PI3K/AKT信号通路有关。Objective： To explore the effects of Achyranths Bidentatae Radix （AB） extracts on monosodium iodoacetate （MIA） -induced injury in rat chondrocytes. Method： Chondrocytes were isolated from the knee joints of Sprague-Dawley rats by using enzymatic digestion. The experiment was divided into 5 groups： the normal group, the model group （4 μmol·L-1 MIA）, the low-, medium-and high-dose AB extract groups （10, 30, 100 mg·L -1 ）. The viability of chondrocytes was detected by MTT assay. The morphology change of chondrocytes was detected by Hoechst 33258 method. The activity of Caspase-3 chondrocytes was measured by spectrophotometry. The activation of AKT and down-stream molecules including Bax, Bcl-2 was assayed by Western Blot. Result： Compared with the normal group, the viability of chondrocytes declined, chondrocytes nucleus shrivel or cracking occurred, the activity of Caspase-3 and the level of Bax protein raised, the level of Bcl-2 protein and phosphorylation of AKT declined in the model group （P 〈 O. O1 ）. Compared with the model group, the viability and morphology of chondrocytes were restored, the expression of Bax was inhibited in the medium-and high-dose AB extract groups （P 〈 0.01 ）. Moreover, the activity of Caspase-3 was inhibited, the expression of Bcl- 2 and the phosphorylation of AKT were promoted in the low-, medium-and high-dose AB extract groups （P 〈 0.01）. Conclusion： These results suggested AB extract exerted anti-apoptosis effect on MIA-induced rat chondrocytes, which might be related to PI3K/AKT signal pathway.

关 键 词：牛膝提取物 骨关节炎 软骨细胞 凋亡 

分 类 号：R285.5[医药卫生—中药学]