骨碎补总黄酮在肿瘤坏死因子α介导下对成骨细胞凋亡的影响

目的：通过观察骨碎补总黄酮在肿瘤坏死因子α（tumor necrosis factor-α,TNF-α）介导下对成骨细胞增殖和凋亡的影响,探讨其防治类风湿关节炎骨质疏松症的作用机制。方法：20只雌性Wistar大鼠,随机分为空白对照组（蒸馏水灌胃）和骨碎补总黄酮组（强骨胶囊灌胃）制备血清。骨碎补总黄酮组大鼠每日强骨胶囊给药剂量为75mg,每日给药2次,2mL/次,连续给药3d,末次给药1h后,心脏采血收集血清备用。细胞实验分为4组：空白血清组、骨碎补总黄酮含药血清组、空白血清＋TNF-α组和骨碎补总黄酮含药血清＋TNF-α组。应用噻唑基四唑法测定成骨细胞的增殖,流式细胞仪测定成骨细胞的凋亡率,实时荧光定量聚合酶链式反应法测定Bcl-2和Bax mRNA表达量。结果：与对照血清相比,骨碎补总黄酮含药血清在TNF-α介导下能够促进成骨细胞增殖（P〈0.05）;减少成骨细胞的凋亡（P〈0.05）,增加Bcl-2/Bax比值。结论：骨碎补总黄酮含药血清在TNF-α介导的炎症环境下能够促进成骨细胞增殖,减少成骨细胞凋亡,可能是通过调节Bcl-2/Bax mRNA比例来实现的。

Effects of Drynaria total flavonoids on apoptosis of osteoblasts mediated by tumor necrosis factor-α

Objective： To investigate the influence of Drynana total flavonoids on proliferation and apoptosis of osteoblasts in tumor necrosis factor-α （TNF-α）- mediated medium, so as to explore the mechanism of Drynaria total flavonoids in preventing and treating osteoporosis of rheumatoid arthritis. Methods： Twenty Wistar rats with average weight of （200220） g were randomly divided into two groups： blank control group and Qianggu capsule （Drynaria total flavonoids） group. Rats in Qianggu capsule group were fed with 75 mg Qianggu capsule daily for continuous 3 d. One hour after the last feed, blood samples were collected. The in vitro experiment of four groups was designed= blank control serum group, Drynaria total fiavonoids-containing serum group, blank control serum plus TNF-~ group and Drynaria total flavonoids- containing serum plus TNF-c~ group. Methyl thiazolyl tetrazolium method was used to detect the proliferation of osteoblasts. Flow cytometry was used to detect the apoptosis of osteoblasts and real-time fluorescent quantitative polymerase chain reaction to detect the expressions of Bcl-2 and Bax mRNAs in osteoblasts. Results： Compared with the control serum, Drynaria total flavonoids-containing serum promoted the proliferation and decreased the apoptosis of osteoblasts in TNF-~-mediated inflammatory environment （P 〈 O. 05）, and increased the ratio of Bcl-2 mRNA to Bax mRNA. Conclusion： In TNF-c~-mediated inflammatory environment, Drynaria total fiavonoids can promote the proliferation and decrease the apoptosis of osteoblasts by improving the ratio of Bcl-2 mRNA to Bax mRNA, which may be one of the mechanisms of Drynaria total flavonoids in preventing and treating osteoporosis of rheumatoid arthritis.