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目的既往研究表明辛伐他汀具有促成骨作用,现探讨辛伐他汀对幼鼠骨小梁中骨形成相关因子表达以及BMSCs成骨分化的影响。方法 40只1周龄雄性SD大鼠,体重17.5～19.5 g,随机分为2组,每组20只。实验组于大鼠颈部皮下注射辛伐他汀注射液[5 mg/(kg.d)],对照组同法注射等剂量生理盐水。于注射后1、3周,两组分别脱颈处死10只大鼠,采用免疫组织化学染色检测胫骨上端骨小梁BMP-2、基质金属蛋白酶13(matrix metalloproteinase 13,MMP-13)、VEGF的表达情况。注射后1、3周,两组分别取大鼠双侧股骨,采用全骨髓培养法体外分离培养BMSCs,取第1代BMSCs成骨诱导培养,培养后14 d行ALP染色,培养后21 d采用实时荧光定量PCR检测BMP-2、Runx2、Osterix、Msx2、Dlx3、Dlx5 mRNA的表达,培养后28 d行von Kossa染色。结果免疫组织化学染色示注射后1、3周两组胫骨上端骨小梁BMP-2、MMP-13、VEGF表达,差异均无统计学意义(P>0.05)。成骨诱导培养14 d,注射后1、3周两组ALP染色阳性细胞百分比差异均无统计学意义(P>0.05);成骨诱导培养21 d,实时荧光定量PCR检测示注射后1、3周两组BMP-2、Runx2、Osterix、Dlx3、Dlx5、Msx2 mRNA表达水平比较差异均无统计学意义(P>0.05);成骨诱导培养28 d,注射后1、3周两组钙结节单位面积比差异均无统计学意义(P>0.05)。结论皮下注射辛伐他汀5 mg/(kg.d)后1、3周对幼鼠骨小梁中骨形成相关因子表达以及BMSCs成骨分化无明显影响。
Purpose Previous studies showed that simvastatin can promote bone formation. We investigated the effect of simvastatin on osteogenesis-related factor expression in bone trabeculae and the osteogenic differentiation of BMSCs in young rats. Methods Forty one-week-old male Sprague-Dawley rats weighing 17.5-19.5 g were randomly divided into two groups of 20 rats. The experimental group was subcutaneously injected simvastatin injection [5 mg / (kg · d)] into the neck of rats, and the control group was given the same dose of normal saline. At 1 and 3 weeks after injection, 10 rats were killed by cervical dislocation. The expression of BMP-2, MMP-13 and VEGF in the upper tibia trabeculae were detected by immunohistochemical staining Express the situation. One and two weeks after injection, bilateral femur were isolated from both groups. BMSCs were isolated and cultured in vitro by whole bone marrow culture. Generation 1 BMSCs were induced to osteogenic induction. ALP staining was performed 14 days after culture. The expression of BMP-2, Runx2, Osterix, Msx2, Dlx3 and Dlx5 mRNA were detected by real-time fluorescence quantitative PCR. The von Kossa staining was performed on the 28th day after culture. Results Immunohistochemical staining showed that the expression of BMP-2, MMP-13 and VEGF in the upper tibia trabeculae of the two groups had no significant difference at 1 and 3 weeks after injection (P> 0.05). There was no significant difference in the percentages of ALP staining positive cells between the two groups after 14-day osteogenic induction culture (P <0.05). The percentage of ALP-positive cells in osteogenic induction group was not significantly different at 1 and 3 weeks after injection (P> 0.05) The expressions of BMP-2, Runx2, Osterix, Dlx3, Dlx5 and Msx2 mRNA in the two groups were not significantly different between the two groups (P> 0.05). After osteogenic induction for 28 days, There was no significant difference in unit area ratio (P> 0.05). Conclusions 1,3 and 3 weeks after simvastatin 5 mg / (kg · d), there is no significant effect on bone formation-related factor expression and osteogenic differentiation of trabecular bone in young rats.