防己诺林碱对糖皮质激素诱导雄性骨质疏松大鼠影响的研究

Effect of fangchinoline on glucocorticoid-induced osteoporosis in male rats

目的探讨防己诺林碱对骨质疏松症的保护作用并假设可能的作用机制。方法通过对大鼠皮下注射泼尼松龙(2.5mg/kg)4周诱导形成骨质疏松症。分别对大鼠腹腔注射防己诺林碱1、3、10 mg/kg,持续一个月时间。通过测量大鼠股骨干骺端组织的微结构参数和骨密度(bone mineral density,BMD)来评估防己诺林碱的保护作用,并检测泼尼松龙诱导的骨质疏松症大鼠血清中的生化指标。同时,测量大鼠股骨组织中Beclin-1、骨形态发生蛋白2(BMP2)、自噬相关5(ATG-5)、Runt相关转录因子2(RUNX-2)和核因子κB配体受体激活因子(RANKL)的基因表达[通过反转录-聚合酶链反应(RT-PCR)蛋白]。结果与泼尼松龙组相比,防己诺林碱治疗组大鼠椎骨组织中BMD和微结构参数显著增加(P<0.01)。同时还发现用防己诺林碱治疗减弱了泼尼松龙诱导的骨质疏松症大鼠的BMP2、Beclin-1、ATG-5、RUNX-2和RANKL蛋白的表达。此外,防己诺林碱治疗和泼尼松龙诱导的骨质疏松症大鼠血清中生化参数水平减弱。结论防己诺林碱通过增加自噬来预防泼尼松龙诱导的骨质疏松症大鼠骨质流失。

Objective The present investigation evaluated the protective effect of fangchinoline against osteoporosis and also postulated the possible mechanism of action.Methods Osteoporosis was induced by subcutaneously injecting prednisolone(2.5 mg/kg)for 4 weeks.Fangchinoline 1,3 and 10 mg/kg was given intraperitoneally for the period.Protective effects of fangchinoline were assessed by estimating microarchitectural parameters and bone mineral density(BMD)of the femoral metaphysis tissues,and biochemical parameters were also determined in the serum of rats with prednisolone-induced osteoporosis.Moreover,gene expression of Beclin-1,bone morphogenetic protein 2(BMP2),autophagy-related 5(ATG-5),Runt-related transcription factor 2(RUNX-2),and receptor activator of nuclear factor kappa-b ligand(RANKL)protein in the vertebrae tissue were assessed by reverse transcription-polymerase chain reaction(RT-PCR)assay.Results There was a significant(P<0.01)increase in the BMD and microarchitectural parameters in the vertebrae tissue of the fangchinoline-treated group compared to the prednisolone group.We also found that treatment with fangchinoline attenuated the altered expressions of BMP2,Beclin-1,ATG-5,RUNX-2 and RANKL protein in the prednisolone-induced osteoporosis rats.Moreover,levels of biochemical parameters were attenuated in the serum of fangchinoline-treated and prednisolone induced osteoporosis rat.Histopathology revealed that the apoptosis of osteoblasts was decreased in the fangchinoline-treated group compared with the prednisolone group of rats.Conclusion Fangchinoline protects against bone loss in prednisolone-induced osteoporosis rats by inducing autophagy.