染料木黄酮对糖尿病大鼠骨微结构的影响及其机制初探 ; Effect of genistein on bone microstructure in femoral head of diabetic rats

目的探讨糖尿病大鼠股骨头骨质疏松倾向变化及染料木黄酮的保护作用。方法选取清洁级4月龄成年雄性SD大鼠60只，体重（200±20）g，按随机数字表法分为3组：正常对照组（CON组）、糖尿病组（DM组）、染料木黄酮干预组（Gen组），每组20只。对DM组和Gen组予链脲佐菌素(STZ)60 mg/kg左下腹腔内注射，建立速发型STZ糖尿病大鼠模型，72 h后取尾血检测血糖，凡血糖≥167 mmol/L即为糖尿病建模成功。两组均完全成模。Gen组予染料木黄酮30 mg·kg?1·d?1，1次/d，CON组注射等体积的枸橼酸缓冲液，均腹腔注射，15周后观察各组股骨头软骨下区以及FⅧ因子免疫组化表达情况。墨汁灌注行微血管相对面积(MVD)测定，分析原位杂交进行的血管内皮生长因子（VEGF） mRNA表达强度。采用最小显著差异法检验及方差分析进行统计学分析。结果（1）光镜下CON组骨小梁粗厚、有完整的成骨细胞区，DM组骨小梁稀疏细薄、细胞核固缩，Gen组骨小梁排列较紧密。（2）与正常对照组比较，DM组成骨细胞减少[（13.2±1.3）比(22.4±2.5)个]，微血管相对面积增大[（8.7±0.7）%比（4.7±0.6）%]，FⅧ因子表达灰度值（137±5比164±5）及VEGF mRNA下降（186±5比213±4），出现骨质疏松倾向。（3）与DM组比较，Gen组骨质疏松倾向及微血管病变改善，成骨细胞增多[（19.2±1.9）个]、微血管单位面积降低[（5.6±0.5）%]，FⅧ因子表达灰度值[（159±7）]及VEGF mRNA表达（200±2）上升，三组间差别具有统计学意义（F=39.191~58.815，均P<0.01）。结论糖尿病早期出现骨质疏松倾向变化，染料木黄酮对糖尿病骨微结构具有保护作用。 ; Objective To assess the osteoporotic changes in diabetic rats and protective effects of genistein. Methods A total of 60 adult SD rats (four months old and weighted (200 ± 20) g) were divided into three groups according to the random number table: normal control group(group CON), diabetic group (Group DM) and intervention group (Group Gen) (20 rats in each group). Rats in Group DM and Group Gen were injected with streptozotocin(STZ) at 60 mg/kg in the left lower abdominal cavity to establish STZ?induced diabetic model, which was considered valid if blood glucose was greater than or equal to 167 mmol/L after 72 hours. Group Gen received 30 mg·kg?1·d?1 of genistein, once a day. An equal volume of citrate buffer was given to Group CON, both were administered intraperitoneally. The immunohistochemical evaluation of the femoral head subchondral region and FⅧ factor were conducted in each group after 15 weeks. The microvessel density was measured with the ink perfusion. Vascular endothelial growth factor(VEGF) mRNA expression was measured and analyzed by the in situ hybridization. The date analysis was performed using LSD test. Results (1) The trabecular bone in Group CON was thick and had complete osteoblasts area. The trabecular bone in Group DM was sparse and thin, ...