EGCG对去势大鼠骨量丢失的预防效果和可能的机制研究

发布时间：2018-05-01 10:28

本文选题：EGCG + 骨质疏松　；参考：《苏州大学》2014年硕士论文

【摘要】：目的：探讨表没食子儿茶素没食子酸酯（(-)-Epigallocatechin-3-gallate, EGCG）对雌性去势（Ovariectomized, OVX）大鼠骨量丢失的早期预防效果、腹腔注射与灌胃途径给药效果和探讨其预防骨量丢失的可能的作用机制。方法：将55只3月龄的正常健康雌性SD大鼠（Sprague-Dawley Rats）分别称重，并按照各组间体重无统计学差异分为以下5组：A组，共15只：去势术后第3天开始以10mg/kg/day的剂量经腹腔（intraperitoneal，IP）注射给药EGCG连续12周，记为（OVX+early EGCG/IP）、B组，共10只:去势术后第7周开始以10mg/kg/day的剂量经腹腔注射EGCG连续给药6周，记为（OVX+late EGCG/IP）、C组，共10只：去势术后第3天开始以10mg/kg/day的剂量经灌胃（Oral Feeding， OF）给药EGCG连续12周，记为（OVX+early EGCG/OF）、D组，共10只：去势大鼠无EGCG干预空白对照组，记为（OVX），和E组，共10只：假手术（Sham-operated）正常对照组，记为（SHAM）。12周后全部予以过量麻醉处死，留取双侧股骨和胫骨标本分别进行如下检测：1、Micro CT扫描，计算骨密度（BMD）和相关骨形态学参数，2、进行3点弯曲实验检测、3、骨组织HE染色，并采用免疫组织化学染色检测Sema4D蛋白的表达情况。结果：骨形态学参数：SHAM组（E组）中大鼠的骨密度（BMD）与松质骨骨体积分数（BV/TV）明显高于其他4组大鼠的BMD以及BV/TV（P0.01），同时A组大鼠的BMD和BV/TV明显高于OVX组（D组）大鼠的BMD（P0.05）。SHAM组（E组）中大鼠的松质骨平均小梁分离度（Tb.Sp）明显低于其余4组的Tb.Sp，且A组大鼠的Tb.Sp明显低于OVX组（D组）大鼠的Tb.Sp（P0.05）。SHAM组（E组）的平均骨小梁厚度（Tb.Th）明显高于其余各组（P0.01），其余4组间无明显差异。各组间松质骨骨小梁数量(Tb.N)无明显差异（P0.05）。3点弯曲应力试验结果：弯曲位移、最大弯曲应力在各组之间，差异均无统计学意义（P0.05）。骨组织学观察：SHAM组（E组）的大鼠骨小梁分布更加致密，，骨小梁数目更多，而OVX组（D组）的大鼠骨小梁稀疏。A组、B组以及C组的组织学形态介于OVX组和SHAM组之间。免疫组化结果：OVX组（D组）的Sema4D的阳性表达面积百分比明显高于其他各组的Sema4D表达水平（P0.05），SHAM组（E组）与A组之间，Sema4D的阳性表达面积百分比差异无统计学意义（P0.05）。结论： 1. EGCG能够延缓雌性去势大鼠骨量丢失的进程，且在10mg/kg/day的剂量下，较早期的干预可能优于较晚期的干预，但是EGCG的早期干预并不能完全阻断去势大鼠骨量丢失的进程； 2.在10mg/kg/day的剂量下，EGCG预防雌性去势大鼠骨量丢失，经腹腔注射途径效果可能优于经灌胃途径； 3.去势大鼠骨组织中的Sema4D表达高于正常大鼠，且EGCG能一定程度降低去势大鼠骨组织中Sema4D的表达。
[Abstract]:Objective: To investigate the early prevention effect of epigallocatechin-3-gallate (EGCGG) on bone mass loss in female ovariectomized (OVXed) rats, the effect of intraperitoneal injection and gavage administration and the possible mechanism of preventing bone loss. Methods: Fifty-five healthy female Sprague-Dawley Ratsrats aged 3 months were weighed and divided into 5 groups: group A according to no significant difference in body weight. A total of 15 rats were given EGCG by intraperitoneal injection of EGCG at the dose of 10mg/kg/day 3 days after ovariectomy for 12 weeks, which was recorded as OVX early EGCG / IP / IP group B (n = 10): from the 7th week after ovariectomy, the dosage of 10mg/kg/day was injected intraperitoneally with EGCG for 6 weeks, which was recorded as OVX late EGCG / IP / IP group C (OVX late EGCG / IP / IP group). Ten rats were treated with oral feeding (10mg/kg/day) for 12 weeks at the dose of 10mg/kg/day on the 3rd day after ovariectomy. The rats were recorded as OVX early EGCG / OFX D group. 10 ovariectomized rats were treated as OVX early EGCG / OFX group without EGCG intervention, as control group, and E group, OVX group, and OVX group (n = 10), and OVX group (n = 10). A total of 10 rats (sham-operated) normal control group, recorded as Sham-operated control group, were all sacrificed under excessive anesthesia after 12 weeks. The specimens of bilateral femur and tibia were taken for the following detection: 1: 1 Micro CT scan, respectively. Bone mineral density (BMD) and related bone morphological parameters were calculated. The expression of Sema4D protein was detected by three point bending test and HE staining in bone tissue. Results: The bone mineral density (BMD) and cancellous bone volume fraction (BV / TVV) of the rats were significantly higher than those of the other four groups (BMD and BV / TVP 0.01), and the BMD and BV/TV of group A were significantly higher than those of BMD(P0.05).SHAM (group D). ) the mean trabecular separation degree of cancellous bone in rats was significantly lower than that in the other four groups, and the Tb.Sp of group A was significantly lower than that of group OVX. (group D) the average trabecular thickness of group Tb.Sp(P0.05).SHAM was significantly higher than that of other groups (P 0.01). There was no significant difference among the four groups. There was no significant difference in the number of trabeculae of cancellous bone in each group. The results showed that the bending displacement and maximum bending stress were not significantly different among the groups. The bone trabecular density and the number of bone trabeculae were more compact in group E than those in group D (OVX group). The histomorphology of group A and group C were between OVX group and SHAM group. Immunohistochemical results showed that the percentage of positive area of Sema4D expression in group D was significantly higher than that in group E (P 0.05). There was no significant difference between group A and group A in the percentage of positive expression of Sema4D (P 0.05). Conclusion: 1. EGCG could delay the process of bone loss in female ovariectomized rats, and at the dose of 10mg/kg/day, the earlier intervention might be better than the late intervention, but the early intervention of EGCG could not completely block the process of bone loss in ovariectomized rats. 2. At the dose of 10mg/kg/day, the effect of intraperitoneal injection may be better than that of gavage in preventing bone loss in female ovariectomized rats. 3. The expression of Sema4D in bone tissue of ovariectomized rats was higher than that of normal rats, and EGCG could decrease the expression of Sema4D in bone tissues of ovariectomized rats to some extent.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R580

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