乳铁蛋白对去卵巢大鼠骨量及骨组织IGF-I、IGFBP-3和IGFBP-5表达的影响

发布时间：2018-11-17 08:59

【摘要】：目的:本研究通过观察乳铁蛋白(Lactoferrin LF)对去卵巢大鼠骨量的影响,以及对股骨IGF-I、IGFBP-3和IGFBP-5 m RNA表达的影响,来探究乳铁蛋白对骨质疏松的作用及其可能的分子机制。方法:将70只6月龄的雌性SD(Sprague-Dawley)大鼠按1:6比例随机分为假手术组(Sham)与去卵巢模型组(Ovx),再将去卵巢大鼠随机平均分为6组:骨质疏松模型组(Con)(生理盐水2ml/只·天,灌胃)、雌激素治疗组(E2)(0.1mg/kg·周,肌肉注射)、牛血清白蛋白组(BSA)(0.1g/kg·d,灌胃)和不同剂量的乳铁蛋白治疗组LF1(0.1g/kg·d,灌胃)、LF2(1g/kg·d,灌胃)、LF3(2g/kg·d,灌胃)。连续干预24周后,采用生物光学显微镜观察大鼠左侧胫骨骺线下干骺端骨组织HE染色病理切片并用图像分析软件(Image-Pro Plus 6.0)分析各组骨小梁平均面积所占骨髓腔比例,再结合实时荧光定量聚合酶链反应(Real Time RT-PCR)来观察大鼠右侧股骨胰岛素样生长因子I(IGF-I)、胰岛素样生长因子结合蛋白3(IGFBP-3)与胰岛素样生长因子结合蛋白5(IGFBP-5)m RNA的表达情况。结果:1.各组大鼠在乳铁蛋白干预前,体重无明显差异(均P0.05),大鼠去卵巢手术后,各组体重均有所增加,以Con组增加最快,乳铁蛋白干预6个月后,各组大鼠体重均明显升高,以Con组增加最多,差别有统计学意义(均P0.05),Con组大鼠明显重于Sham组和E2组(均P0.05),BSA组和不同剂量乳铁蛋白治疗组大鼠体重均低于Con组,但无统计学意义(均P0.05)。2.各组大鼠治疗6个月后,显微镜下观察骨组织HE染色病理切片发现Sham组和E2组较Con组和BSA组的骨小梁更为致密,LF1-3(0.1g/kg·d,1g/kg·d,2g/kg·d)骨小梁较Con组也更为致密,但不如Sham组和E2组显著。LF1-3各组之间骨小梁的疏密程度在肉眼观差异不大。图像分析提示骨小梁面积占骨髓腔的比例Sham组和E2组明显高于Con组、BSA组和LF1-3各组(均P0.01),Sham组和E2组、BSA组和Con组、LF1-3各组之间均无明显差异(均P0.05)。3.各组大鼠治疗6个月后,Con组IGF-I m RNA表达量显著低于Sham组和E2组(均P0.01),也低于LF1-3(0.1g/kg·d,1g/kg·d,2g/kg·d)各组,差别有统计学意义(均P0.01)。Con组IGFBP-3 m RNA表达量明显低于Sham组和E2组(P0.01),但高于LF1-3(0.1g/kg·d,1g/kg·d,2g/kg·d)各组,差别有显著性(均P0.01)。Con组IGFBP-5 m RNA表达量显著低于Sham组和E2组(均P0.01),也低于LF2-3(1g/kg·d,2g/kg·d)各组,差别有统计学意义(P0.05和P0.01)。结论:1.成功建立去卵巢大鼠的骨质疏松模型。2.乳铁蛋白可提高去卵巢大鼠的骨量,减少因去卵巢而诱发的骨丢失,这可能对骨质疏松的治疗有潜在作用。3.乳铁蛋白改善骨质疏松可能通过提高IGF-I和IGFBP-5水平,抑制IGFBP-3水平,从而促进骨形成,抑制骨吸收。
[Abstract]:Objective: to observe the effect of lactoferrin (Lactoferrin LF) on bone mass and the expression of IGF-I,IGFBP-3 and IGFBP-5 m RNA in femur of ovariectomized rats. To explore the role of lactoferrin in osteoporosis and its possible molecular mechanisms. Methods: seventy 6-month-old female SD (Sprague-Dawley) rats were randomly divided into sham operation group (Sham) and ovariectomized model group (Ovx),) at 1:6. Then the ovariectomized rats were randomly divided into 6 groups: osteoporosis model group (Con) (saline 2ml/, intragastric perfusion), estrogen treatment group (0.1mg/kg week, intramuscular injection), ovariectomized rats (ovariectomized rats), ovariectomized rats (ovariectomized rats). Bovine serum albumin (BSA) group (BSA) (0.1g/kg d) and different doses of lactoferrin treatment group (0.1g/kg d, gavage), LF2 (1g/kg d, gavage), LF3 (2g/kg d, gavage). After 24 weeks of continuous intervention, the HE staining pathological sections of the left tibial epiphyseal bone of the rats were observed under a biomicroscopy and the ratio of the average trabecular area to the medullary cavity in each group was analyzed by image analysis software (Image-Pro Plus 6.0). The right femur insulin-like growth factor I (IGF-I) was observed by real-time fluorescence quantitative polymerase chain reaction (Real Time RT-PCR). Expression of insulin-like growth factor binding protein 3 (IGFBP-3) and insulin-like growth factor binding protein 5 (IGFBP-5) m RNA). The result is 1: 1. Before lactoferrin intervention, there was no significant difference in body weight in each group (P0.05). After ovariectomy, the body weight of each group increased, especially in Con group. After 6 months of lactoferrin intervention, the body weight of each group increased significantly. The increase of Con group was the most, and the difference was statistically significant (P0.05), Con group rats were significantly heavier than Sham group and E2 group (P0.05), BSA group and different doses of lactoferrin treatment group) rats were lower than Con group. However, there was no statistical significance (P0.05). 2. After 6 months of treatment, the pathological sections of HE staining of bone tissue were observed under microscope. The bone trabeculae of Sham group and E2 group were denser than those of Con group and BSA group, LF1-3 (0.1g/kg dU 1 g / kg / kg d). The trabecular density of 2g/kg d was also more compact than that of Con group, but it was less significant than that of Sham group and E2 group. The degree of trabecular density in LF1-3 group was not significantly different from that in Con group. The ratio of trabecular area to medullary cavity in Sham group and E2 group was significantly higher than that in Con group, BSA group and LF1-3 group (P0.01), Sham and E2 group, BSA group and Con group). There was no significant difference between LF1-3 groups (P0.05). After 6 months of treatment, the expression of IGF-I m RNA in Con group was significantly lower than that in Sham group and E2 group (P0.01), and also lower than that in LF1-3 group (1 g / kg 0.1g/kg dai 2g / kg d). The expression of IGFBP-3 m RNA in the P0.01). Con group was significantly lower than that in the Sham group and E2 group (P0.01), but higher than that in the LF1-3 (0.1g/kg 1 g / kg / kg d ~ (2) g / kg) group. The expression of IGFBP-5 m RNA in P0.01). Con group was significantly lower than that in Sham group and E2 group (P0.01), and was also lower than that in LF2-3 (2 g / kg 1g/kg d) group. The difference was statistically significant (P0.05 and P0.01). Conclusion: 1. Successful establishment of osteoporosis model in ovariectomized rats. 2. Lactoferrin can increase bone mass and reduce bone loss induced by ovariectomy in ovariectomized rats, which may play a potential role in the treatment of osteoporosis. The improvement of osteoporosis by lactoferrin may promote bone formation and inhibit bone resorption by increasing the levels of IGF-I and IGFBP-5 and inhibiting the level of IGFBP-3.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R580

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