左旋紫草素自组装纳米粒对破骨细胞分化的抑制作用研究

Inhibitory effect of L-shikonin loaded self-assembled nanoparticles on osteoclast differentiation

目的探讨左旋紫草素自组装(Asp)6-PEI@SK-PLGA纳米粒在体外对破骨细胞分化的抑制作用。方法体外培养小鼠原代外周血单核细胞(PBMCs),72 h诱导分化为破骨细胞,分为空白对照组(培养基不含左旋紫草素)、左旋紫草素(Lshikonin,SK)原料药组(20μg/L)、(Asp)6-PEI@SK-PLGA纳米粒组(20μg/L),连续给药5 d。抗酒石酸酸性磷酸酶(TRAP)染色观察破骨细胞形态、实时荧光定量逆转录聚合酶链反应(qRT-PCR)检测破骨细胞特异基因表达。结果成功诱导PBMCs细胞为破骨细胞,与SK原料药组相比,(Asp)6-PEI@SK-PLGA组TRAP染色阳性细胞数显著降低且面积显著减少(P <0.05),NFATc1、c-fos mRNA水平显著降低(P <0.05)。结论相对于原料药组,(Asp)6-PEI@SK-PLGA纳米粒能显著抑制破骨细胞分化。

Objective To explore the in vitro inhibitory effect of L-shikonin loaded self-assembled nanoparticles(Asp)6-PEI@SK-PLGA on osteoclast differentiation. Methods Primary peripheral blood mononuclear cells(PBMCs) of mice were cultured in vitro and induced into osteoclasts at 72 h. The osteoclasts were divided into a blank control group(no L-shikonin in the medium), a L-shikonin SK active substance group(20 μg/L) and a(Asp)6-PEI@SK-PLGA nanoparticles group(20 μg/L). All these groups were administered continuously for five days. Tartrate resistant acid phosphatase(TRAP) staining was used to observe the morphology of osteoclasts and real time quantitative reverse transcription polymerase chain reaction(qRT-PCR) was used to detect specific gene expression of osteoclasts. Results PBMCs were induced into osteoclasts successfully, compared with SK group, the number and the area of TRAP positive cells were significantly decreased in(Asp)6-PEI@SK-PLGA group(P < 0.05), and the mRNA levels of NFATc1 and c-fos were significantly decreased(P < 0.05). Conclusion Compared with SK group,(Asp)6-PEI@SK-PLGA nanoparticles can significantly inhibit osteoclast differentiation.