益气化瘀补肾方血清对兔关节软骨细胞增殖与Caspase3/7酶活性的影响  

Effects of "Yiqi Huayu Bushen Decoction" serum on articular chondrocytes proliferation and Caspase3/7 activity in rabbits

作　　者：胡波[1] 唐德志[1] 赵永见[1] 张有为[2] 周泉[1] 胡志俊[2] 施杞[1] 王拥军[1] 

机构地区：[1]上海中医药大学脊柱病研究所,上海200032 [2]上海中医药大学附属龙华医院,上海200032

出　　处：《上海中医药杂志》2010年第12期73-75,共3页Shanghai Journal of Traditional Chinese Medicine

基　　金：上海市科委基础重点项目(08JC1418700);上海市科委基础研究重点项目(07JC14050)

摘　　要：目的观察益气化瘀补肾方含药血清对体外培养的兔关节软骨细胞增殖与Caspase3/7酶活性的影响。方法取兔的膝关节软骨,将软骨细胞分离传代后,取第三代细胞,用10μg/L的IL-1β诱导正常软骨细胞退变,制备益气化瘀补肾方含药血清作为干预措施。益气化瘀补肾方组分别加入IL-1β及不同溶度(5%,10%,20%)益气化瘀补肾方含药血清培养液,对照组分别加入IL-1β及不同溶度(5%,10%,20%)空白血清培养液,分别培养24h、48h后,采用MTT比色法检测软骨细胞增殖。另取对数生长期第三代软骨细胞分为4组:正常软骨细胞+20%空白血清;正常软骨细胞+20%益气化瘀补肾方含药血清;IL-1β诱导软骨细胞+20%空白血清;IL-1β诱导软骨细胞+20%益气化瘀补肾方含药血清。每组6个复孔,每孔100μl,各组分别用相应的血清培养24h、48h后进行检测,采用Caspase3/7试剂盒检测各组软骨细胞的Caspase3/7酶活性变化情况。结果不同浓度的益气化瘀补肾方血清均能拮抗IL-1β抑制软骨细胞增殖;与正常软骨细胞+20%空白血清组比较,IL-1β诱导软骨细胞+20%空白血清组Caspase3/7酶活性升高,差异有统计学意义(P<0.05);与IL-1β诱导软骨细胞+20%空白血清组比较,IL-1β诱导软骨细胞+20%益气化瘀补肾方血清组Caspase3/7酶活性降低,差异有统计学意义(P<0.05);正常软骨细胞+20%益气化瘀补肾方血清组的Caspase3/7酶活性与正常软骨细胞+20%空白血清组和IL-1β诱导软骨细胞+20%益气化瘀补肾方血清组差异无统计学意义(P>0.05)。结论益气化瘀补肾方血清能拮抗IL-1β抑制软骨细胞增殖,下调软骨细胞Caspase3/7酶活性,从而抑制软骨细胞的凋亡。Objective To observe the effects of ＂Yiqi Huayu Bushen Decoction＂ serum on articular chondrocytes proliferation and Caspase3/7 activity in rabbits.Methods Take the rabbit knee cartilage and separate the chondrocytes nestin.The third generation cells were used to induce cartilage cells degeneration with 10 μg/L speeds of IL-1beta to make ＂Yiqi Huayu Bushen Decoction＂ serum as intervention measure.In observation groups,IL-1beta and different concentrations of ＂Yiqi Huayu Bushen Decoction＂（5%,10%,20%） were added;and in control group,IL-1beta and different concentrations（5%,10%,20%） of serum-free were added.MTT colorimetric method was adopted to detect chondrocytes proliferation.The third generation of trypanblau growth chondrocytes were then divided into four groups：group A with normal chondrocytes and 20% serum-free,group B with normal chondrocytes and 20% ＂Yiqi Huayu Bushen Decoction＂ serum,group C with IL-1beta-induced chondrocytes and 20% serum-free,group D with IL-1beta-induced chondrocytes and 20% ＂Yiqi Huayu Bushen Decoction＂ serum.In each group,six double holes with 100μl serum were cultured for 24 h and 48 h,and the Caspase3/7 kit was used to assay the Caspase3/7 activity.Results Different concentrations of ＂Yiqi Huayu Bushen Decoction＂ serum could inhibit IL-1beta-induced chondrocytes proliferation.Compared with group A,the caspase3/7 activity was increased in group C（P0.05）;compared with group B,the caspase3/7 activity was increased in group D.There was no difference in the Caspase3/7 activity between groups A,B and D（P0.05）.Conclusion＂Yiqi Huayu Bushen Decoction＂ can inhibit IL-1beta-induced chondrocytes proliferation by down-regulating Caspase3/7 to suppress chondrocytes apoptosis.

关 键 词：骨关节炎 软骨细胞增殖 益气化瘀补肾方 血清药理学 Caspase3/7 

分 类 号：R285.5[医药卫生—中药学]