白藜芦醇抑制大鼠生长板软骨细胞凋亡及对胫骨生长的影响  

Effect of resveratrol on growth plate chondrocyte apoptosis and tibia growth in rats

作　　者：郭畅 谢艳 李冬冬 殷娜 张丽 GUO Chang;XIE Yan;LI Dongdong;YIN Na;ZHANG Li(Luoyang Orthopedic-Traumatological Hospitul of Henan Province(Henan Provincial Orthopedic Hospital),Luoyang,Henan 471002,China)

机构地区：[1]河南省洛阳正骨医院(河南省骨科医院),河南洛阳471002

出　　处：《上海中医药杂志》2021年第12期82-87,共6页Shanghai Journal of Traditional Chinese Medicine

基　　金：河南省中医药科学研究专项课题(2018ZY1023)。

摘　　要：目的探索白藜芦醇对IL-1β诱导的生长板软骨细胞凋亡的影响,以及筛选白藜芦醇作用的最适浓度,并对白藜芦醇延迟大鼠骨骺闭合、促进长骨生长的作用进行验证。方法取4周龄SD大鼠,对其生长板软骨细胞进行分离及培养,Ⅱ型胶原免疫荧光实验进行细胞鉴定。将软骨细胞随机分为9组:正常对照组(普通培养基)、模型对照组(含20 ng/mL IL-1β)、阳性对照组(含20 ng/mL IL-1β+5μmol/L来曲唑)、二甲基亚砜(DMSO)对照组(培养基里加入白藜芦醇的溶解物DMSO)、实验1组(只加入40μmol/L白藜芦醇)、实验2组至实验5组(分别含20 ng/mL IL-1β+10、20、40、60μmol/L白藜芦醇)。MTT比色法筛选白藜芦醇抑制生长板软骨细胞凋亡、促进细胞增殖的最适浓度,流式细胞仪检测最适浓度的白藜芦醇抑制软骨细胞凋亡情况;并用适量浓度的白藜芦醇给予大鼠灌胃,观察其对大鼠胫骨平台骨骺闭合及胫骨生长的影响。结果实验分离培养获得大鼠生长板软骨细胞。MTT法检测结果显示,与正常对照组比较,模型对照组细胞增殖能力显著降低(P<0.05)。与模型对照组比较,来曲唑和不同浓度白藜芦醇均能拮抗IL-1β引起的细胞凋亡,差异有统计学意义(P<0.01),白藜芦醇的干预效果优于来曲唑,且最佳浓度为40μmol/L。流式分析结果显示,模型对照组细胞凋亡率显著高于正常对照组(P<0.05),40μmol/L的白藜芦醇能够降低IL-1β诱导的大鼠生长板软骨细胞的凋亡比例,且干预效果优于阳性对照组来曲唑。100 mg/kg给予大鼠灌胃白藜芦醇可以延迟大鼠胫骨平台骨骺闭合且促进胫骨生长。结论40μmol/L白藜芦醇能明显抑制IL-1β诱导的软骨细胞凋亡,而100 mg/kg白藜芦醇给予大鼠灌胃可以延迟大鼠胫骨平台的骨骺闭合,为骨骼生长争取更长时间从而促进发育期骨骼增长。Objective To explore the effect of resveratrol on IL-1β induced apoptosis of growth plate chondrocytes, and to screen the optimal concentration of resveratrol and verify the effect of resveratrol on delaying the closure of the rat epiphysis and promoting the growth of long bones. Methods The growth plate chondrocytes of 4-week-old SD rats were isolated and cultured. The cells were identified by type Ⅱ collagen immunofluorescence. The chondrocytes were randamly divided into 9 groups: normal control group(ordinary medium), model control group(containing 20 ng/mL IL-1β), positive control group(containing 20 ng/mL IL-1β and 5 μmol/L letrozole), DMSO control group(DMSO added with resveratrol in culture medium), experimental group 1(only 40 μmol/L resveratrol), experimental group 2 to experimental group 5(containing 20 ng/mL IL-1 β and 10, 20, 40, 60 μmol/L resveratrol, respectively). MTT colorimetric method was used to screen the optimal concentration of resveratrol to inhibit chondrocyte apoptosis and promote cell proliferation. Flow cytometry was used to detect the optimal concentration of resveratrol to inhibit chondrocyte apoptosis. The effects of resveratrol on epiphyseal closure and tibial growth of tibial plateau in rats were observed by intragastric administration of appropriate concentration of resveratrol. Results Chondrocytes of growth plate were isolated and cultured. MTT assay results showed that compared with the normal control group, the proliferation ability of the model control group was significantly decreased(P<0.05);compared with the model control group,letrozole and resveratrol at different concentrations could antagonize the apoptosis induced by IL-1β,and the difference was statistically significant( P < 0. 01). The intervention effect of resveratrol was better than letrozole,and the optimal concentration was 40 μmol/L. The results of flow cytometry showed that the apoptosis rate of the model control group was significantly higher than that of the normal control group( P < 0.05). Resve

关 键 词：白藜芦醇 软骨细胞 凋亡 来曲唑 胫骨 大鼠 中药研究 

分 类 号：R285.5[医药卫生—中药学]