雷洛昔芬对大鼠骨质疏松相关关节退变的作用研究

发布时间：2018-05-12 01:40

本文选题：骨关节炎 + 骨质疏松　；参考：《华北理工大学》2017年硕士论文

【摘要】：目的旨在探讨成年SD大鼠进行双侧卵巢切除术(Ovariectomy,OVX)模拟人类绝经期变化,制造绝经后骨关节炎(Osteoarthritis,OA)模型的可行性;早期应用雷洛昔芬(Raloxifene,RAL)干预绝经后OA模型,研究RAL对软骨下骨及关节软骨的作用;并且观察大鼠年龄相关的关节变化。方法24只6月龄SPF级SD大鼠随机分为4组,每组6只,即:Baseline组、Sham+V组、OVX+V组和OVX+RAL组。Baseline组在实验开始时进行安乐死取材,OVX+V组和OVX+RAL组行双侧OVX手术,Sham+V组进行对照性手术,不进行实质性卵巢切除。术后OVX+RAL组给予RAL 6.2mg/kg·day灌胃,OVX+V组和Sham+V组给予等量生理盐水灌胃。给药3月后安乐死动物,4%甲醛固定液固定。所有标本先应用Micro-CT对右侧近端胫骨行骨组织形态学测定;Micro-CT测定完成后,所有动物右膝关节脱钙8周,石蜡包埋切片,切片行甲苯胺蓝染色和COL-II、Caspase-3,MMP-3免疫组织化学染色。甲苯胺蓝染色切片应用OARSI组织学评分;免疫组织化学阳性蛋白表达采用IRS评分。各组数据进行相应的统计学分析。结果1 OARSI组织学评分发现,OVX+V组分别与Sham+V组和OVX+RAL组比较除总软骨退变宽度和关节囊修复外,所有评分均升高,特别是软骨基质丢失宽度和显著软骨退变宽度显著升高(P0.05),Sham+V组与Baseline组比较除软骨基质丢失宽度和显著软骨退变宽度评分相同外,所有评分均增加,但无统计学差异。2 OVX+V组分别与Sham+V组和OVX+RAL组比较COL-II表达均显著降低(P分别0.001和0.001);Sham+V组与Baseline组比较COL-II表达显著降低(P=0.049)。同样,OVX+V组分别与Sham+V组和OVX+RAL组比较Caspase-3稍显升高,但无统计学差异;Sham+V组与Baseline组比较稍显升高,但无统计学差异。OVX+V组分别与Sham+V组和OVX+RAL组比较MMP-13表达均显著增加(P均为0.000);Sham+V组与Baseline组比较稍显升高,但无统计学差异。OVX+V组分别与Sham+V组和OVX+RAL组比较COL-II/Caspase-3和COLII/MMP-13比值均显著降低(P0.05);Sham+V组与Baseline组比较,COLII/Caspase-3和COL-II/MMP-13比值稍显降低,其中COL-II/MMP-13有统计学差异(P=0.006)。3 Micro-CT发现,OVX+V组与Sham+V组比较,BV/TV、Tb.N、I.S和BS/TV值均降低,其中I.S具有统计学差异(P=0.022),Tb.Sp和SMI值均增加,但无统计差异。OVX+V组与OVX+RAL组比较,BV/TV、Tb.N、I.S和BS/TV值均显著降低(P分别=0.001、0.000、0.000和0.002),Tb.Sp和SMI值均显著增加(P分别=0.012和0.002);Sham+V组与Baseline组比较,BV/TV、Tb.N、I.S和BS/TV值均稍显降低,Tb.Sp和SMI值均稍显升高,但无统计学差异。结论1.OVX诱导的软骨下骨和关节软骨退变较为轻微,提示6月龄OVX大鼠是一种退变较为轻微的绝经后OA大鼠模型。2.RAL经口灌喂3个月可以延缓OVX诱导的绝经后软骨下骨和软骨退变,并且改善OVX所诱导的关节囊改变;关节是一个有机整体,关节组织间相互作用,RAL对关节软骨、软骨下骨、关节囊等关节组织可能都具有积极作用,从关节整体上延缓OA的疾病进展。3.随着成年大鼠年龄的增加,软骨下骨和关节软骨退变有加重趋势。
[Abstract]:Objective to explore the feasibility of Ovariectomy (OVX) in adult SD rats to simulate the changes of human menopause and to make Osteoarthritis (OA) model. Early use of Raloxifene (RAL) to intervene the postmenopausal OA model and study the effect of RAL on the subchondral bone and articular cartilage. Methods 24 6 month old SPF grade SD rats were randomly divided into 4 groups, with 6 rats in each group, that is, group Baseline, group Sham+V, group OVX+V and group OVX+RAL were euthanasia at the beginning of the experiment, OVX+V and OVX+RAL groups underwent bilateral OVX operation, Sham+V group performed controlled surgery without substantial ovarian resection. O after operation Group VX+RAL was given RAL 6.2mg/kg / day for gastric perfusion, and group OVX+V and Sham+V group were given the same amount of normal saline. After March, the animals were euthanized with 4% formaldehyde fixed solution. All specimens were first used to determine the histomorphology of the right proximal tibia with Micro-CT; after Micro-CT, all the animals' right knee joints were decalcified for 8 weeks and paraffin embedded sections. The sections were stained with toluidine blue and immunohistochemical staining of COL-II, Caspase-3 and MMP-3. The histology score of toluidine blue was used in OARSI histology; the expression of immunohistochemical positive protein was evaluated by IRS score. The data of each group were analyzed statistically. The results of 1 OARSI histology were found in the OVX+V group with Sham+V and OVX+RAL groups, respectively. In addition to the total cartilage degeneration width and articular capsule repair, all scores were increased, especially the cartilage matrix loss width and significant cartilage degeneration width increased significantly (P0.05). In group Sham+V and Baseline group, all scores were increased except for the same cartilage matrix loss width and significant cartilage degeneration width score, but no statistical difference was found in.2 group. Compared with group Sham+V and group OVX+RAL, the expression of COL-II was significantly decreased (P 0.001 and 0.001), and the COL-II expression in Sham+V group was significantly lower than that in Baseline group (P=0.049). Similarly, the OVX+V group was slightly higher than the Sham+V group and the OVX+RAL group, but there was no difference between the OVX+V group and the OVX+RAL group, but the group was slightly higher than the Baseline group. Compared with group Sham+V and OVX+RAL group, the expression of MMP-13 was significantly increased in.OVX+V group (P was 0), and in Sham+V group and Baseline group a little increased, but there was no statistical difference between.OVX+V group and Sham+V group and OVX+RAL group. In comparison, the ratio of COLII/Caspase-3 and COL-II/MMP-13 decreased slightly, and COL-II/MMP-13 had statistical difference (P=0.006).3 Micro-CT. The BV/TV, Tb.N, I.S and BS/TV values were all lower in OVX+V group than in Sham+V group. The values of Tb.N, I.S and BS/TV were significantly decreased (P =0.001,0.000,0.000 and 0.002), Tb.Sp and SMI were both significantly increased (P =0.012 and 0.002), and Sham+V and Baseline were slightly lower than those in Baseline group. It changed slightly, suggesting that the 6 month old OVX rat is a mild postmenopausal OA rat model of postmenopausal OA rat model.2.RAL can delay the degeneration of subchondral bone and cartilage induced by OVX, and improve the change of articular capsule induced by OVX; the joint is an organic whole, interarticular intertissue interaction, RAL to articular cartilage and soft tissue. Subosseous bone, joint capsule and other joint tissues may have positive effects. The disease progresses from the joint as a whole, and the disease progression of OA.3. is increased with the age of adult rats, and the degeneration of subchondral bone and articular cartilage is aggravated.

【学位授予单位】：华北理工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R684.3

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