酒精摄入对酒精性骨重构大鼠骨形态、生物力学及骨髓稳态的影响

Effects of alcohol intake on bone morphology,bone biomechanics and bone marrow homeostasis in the rat model of alcoholic bone remodeling

目的探讨长期酒精摄入对酒精性骨重构大鼠模型骨组织形态、骨生物力学、骨髓稳态及氧化应激的影响。方法雄性SD大鼠20只随机分为空白组和酒精组,每组各10只。酒精组大鼠采用20%(vol/vol)酒精腹腔注射,10 mL/kg,每日1次,共12周;正常组给予等体积生理盐水腹腔注射。收集股骨、胫骨和第五椎体行HE和micro-CT扫描评价酒精对骨小梁等骨微观结构的影响;Masson染色评价酒精对胶原纤维蛋白数量及分布的影响;生物力学测试(三点弯曲试验、压缩试验)评价酒精对骨生物力学性能的影响;qRT-PCR法检测骨髓提取及其骨稳态标志物表达情况,评价酒精对COL1A2、BMP2、RANKL和OPG mRNA表达的影响;TBA法/WST-1法检测血清MDA、SOD及SOD/MDA水平。结果与空白组相比,酒精组骨小梁及胶原纤维蛋白数量减少,骨生物力学性能降低;骨髓COL1A2、BMP2和OPG mRNA表达降低,RANKL mRNA表达增加;血清SOD及SOD/MDA水平降低,而MDA水平增加。结论酒精可导致骨小梁及胶原纤维蛋白数量减少、生物力学性能降低、骨髓成骨/破骨功能失衡,破坏了机体氧化还原平衡,导致机体处于高氧化应激状态,增加了罹患骨病风险。

Objective To investigate the effects of long-term alcohol intake on bone tissue morphology,bone biomechanics,bone marrow homeostasis,and oxidative stress in the alcoholic bone remodeling rat model.Methods Twenty male SD rats were randomly divided into a blank group and an alcohol group,with 10 in each group.Rats in the alcohol group were injected intraperitoneally with 20%(vol/vol)alcohol,10 mL/Kg,once a day for 12 weeks.Rats in the normal group were injected intraperitoneally with an equal volume of normal saline.The femur,tibia,and the fifth vertebrae were collected for HE staining and micro-CT scans to evaluate the effect of alcohol on the bone microstructure such as trabecular bone.Masson staining was used to evaluate the effect of alcohol on the amount and distribution of collagen fibers.Biomechanical tests(three-point bending test and compression test)were used to evaluate the effect of alcohol on bone biomechanical properties.qRT-PCR method was used to detect bone marrow extraction and the expression of bone homeostasis markers,and to evaluate the effect of alcohol on COL1A2,BMP2,RANKL,and OPG mRNA expressions.TBA method/WST-1 method was used to detect serum levels of MDA,SOD,and SOD/MDA.Results Compared to those in the blank group,the amount of trabecular bone and collagen fibers in the alcohol group decreased,the biomechanical properties of bone decreased,the expression of bone marrow COL1A2,BMP2,and OPG mRNA decreased,the expression of RANKL mRNA increased,serum SOD and SOD/MDA levels decreased,and the MDA content increased.Conclusion Alcohol reduces the amount of bone trabecula and collagen fibers,decreases the biomechanical properties,induces bone marrow osteogenesis/osteogenesis imbalance,destroys the balance of oxidation/reduction in the body,causes high oxidative stress in the body,and increases the risk of bone diseases.