目的:运用去卵巢大鼠骨丢失动物模型,观察密骨胶囊对骨内Ⅰ型胶原代谢的影响。方法:3月龄清洁级健康雌性Wistar大鼠110只,体质量(120±10)g,采用卵巢切除动物模型,造模后将大鼠随机分为假切组、模型组和密骨胶囊组,分别于给药4周、8周、12周处死大鼠。运用酶联免疫吸附法检测尿脱氧吡啶啉(DPD)、吡啶啉(PYD);运用蛋白印迹免疫法检测骨内Ⅰ型胶原蛋白的表达,运用实时荧光定量聚合酶链式反应检测骨内COLIα1和COLIα2基因的表达及赖氨酸羟化酶2b(LH2b)基因表达。结果:药物干预4周后各组尿PYD/DPD比值比较未见明显差异;而药物干预8周后,模型组尿PYD/DPD比值减小,密骨胶囊组未见明显变化,且与模型组相比差异有统计学意义(P〈0.05)。大鼠去卵巢造模成功后,模型组骨内Ⅰ型胶原蛋白的表达处于低水平,与假切组比较差异有统计学意义(P〈0.05);药物干预4周、8周和12周后,模型组骨内Ⅰ型胶原蛋白表达始终较低,而密骨胶囊组的Ⅰ型胶原蛋白在上述三个时间点的表达均较模型组高,且差异有统计学意义(P〈0.05)。实时PCR结果提示,模型组骨内COL1α1mRNA和COL1α2mRNA的表达明显降低,与假切组比较差异有统计学意义(P〈0.05).干预12周后,密骨胶囊组骨内COL1α1mRNA和COL1α2mRNA表达均明显升高,且均与模型组比较差异有统计学意义(P〈0.05)。去卵巢后大鼠骨内LH2bmRNA表达异常升高,且与假切组比较差异有统计学意义(P〈0.05);密骨胶囊干预后,大鼠显示出降低因去卵巢引起LH2bmRNA表达异常增高的作用,且与模型组比较差异有统计学意义(P〈0.05)。结论:密骨胶囊能上调骨内Ⅰ型胶原的表达,并且通过恢复去卵巢后LH2b异常表达改善Ⅰ型胶原代谢,进而改善骨基质结构。
Objective: To observe the influence of type I collagen metabolism of bone on ovariectomized rat animal model of bone loss when applied with Migu capsule. Methods: All 110 clean level female Wistar rat, three months, weight( 120 - 10)g, were established into ovariectomized rat animal model of bone loss. They were randomly divided into sham group, model group, and Migu capsule group. They were executed at 4th, 8th, and 12th weeks respectively. The urine deoxidization pyridine moiety (DPD) and pyridine moiety (PYD) were detected by enxyme-linked immunosorbent method. The expression of type I collagen in bone were detected by Western blot method. The expression of COLIal, COLIa2, and lysine hydroxylase 2b (LH2b) gene were detected by real-time PCR. Results: After four weeks, the urine PYD/DPD ratio was not significantly different in each groups; after eight weeks, the urine PYD/DPD ratio was decreased in model group, but was not significantly changed in the Migu capsule group, which was significantly different with model group(P〈0. 05). The expression of type I collagen protein was at a lower level in model group at baseline, which was significantly different compared with the sham group(P〈0.05). The expression of type 1 collagen protein was at a lower level in model group at 4th weeks, 8th weeks, and 12th weeks too, which were significantly lower than Migu capsule group respectively(P〈0.05). The expression of COLIal and COLIa2 mRNA were decreased obviously than the sham group(P〈0.05). After 12 weeks, lhe expression of COLIel and COLle2 mRNA were significantly increased in Migu capsule group, which was significantly different with the model group(P〈0.05). The expression of LH2b mRNA were significantly increased in model group, which was significantly different with the sham group(P〈0.05). The expression of LH2b mRNA was significantly different between Migu capsule group and model group(P〈0.05). Conclusion: Migu capsule can increase the expression of ty