摘要
目的:追踪观察移植细胞在骨折愈合不同时间点的神经型一氧化氮合酶、诱导型一氧化氮合酶与内皮型一氧化氮合酶动态表达的生物学意义。方法:实验于1999-01/2004-02在中山大学动物试验中心、中山大学病理教研室完成、中山大学肿瘤防治中心完成。选择SD清洁级大鼠96只,按配对原则分为实验组48只,对照组48只;SD清洁级乳鼠20只进行成骨细胞原代培养。建立SD大鼠老龄骨质疏松骨折的动物模型,实验组将SD乳鼠颅骨体外培养的成骨细胞移植到SD雌性大鼠骨质疏松性骨折部位,对照组注入等量无血清培养液。手术后3,7,10,14,21,28,56,84d处死,每组每个时间点6只。用免疫组化检测骨折愈合不同时相的标本诱导型一氧化氮合酶、内皮型一氧化氮合酶和神经元型一氧化氮合酶的动态表达情况。结果:96只大鼠均进入结果分析。实验组:诱导型一氧化氮合酶在3d左右可见较多阳性表达的细胞,7d有表达高峰。内皮型一氧化氮合酶在3d左右可见少量阳性细胞,14d有表达高峰。神经元型一氧化氮合酶在10d左右可见少量阳性细胞,14～21d表达稍有增高,整个实验过程中神经元型一氧化氮合酶表达较弱,表达高峰不明显。而对照组诱导型一氧化氮合酶、内皮型一氧化氮合酶均有较弱表达,表达峰低平,而神经元型一氧化氮合酶表达很弱,未见明显表达高峰。结论:诱导型一氧化氮合酶、内皮型一氧化氮合酶在成骨细胞移植促进老年骨质疏松性骨折愈合的表达具有时效性,且表达的定位细胞不同,表达的量有差别,提示骨折部位一氧化氮合酶的释放可能对促进骨质疏松性骨折愈合有意义。神经元型一氧化氮合酶在成骨细胞移植促进老年骨质疏松性骨折愈合过程中的作用不明显。
AIM： To study the biological function of neuronal nitric oxide synthase （nNOS）, inducible nitric oxide synthase（iNOS） and endothelial nitric oxide synthase（eNOS） at different time points of the healing of osteoporotic fracture. METHODS： The experiment was conducted in the Animal Experimental Center, the Pathology Department and the Curing and Preventive Center for Tumor of Sun Yat-sen University between January 1999 and February 2004. Ninety-six SD rats were selected and randomized into experimental group（n=48） and control group（n=48） according to the matching method. Another 20 SD neonatal rats were enrolled for primary culture of osteoblasts. After fracture models of osteoporosis rats were established, in vitro cultured osteoblasts from the skull of SD neonatal rats were implanted into the fractured site of female SD rats in the experimental group, while the control group was injected with serum-free medium of the same volume. Each six rats were killed at days 3,7,10,14,21,28,56,84 postoperatively, respectively. Dynamic expression of nNOS,iNOS, and eNOS were measured by using immunohistochemistry at different time phase of fracture healing. RESULTS： All the 96 rats were involved in the result analysis. In the experimental group, . iNOS could be seen positive on day 3 after osteoblast grafts, and iNOS expression reached the peak value on day 7; eNOS could be seen weakly positive on day 3 after osteoblast grafts, and eNOS expression reached the peak value on day 14; eNOS could be seen weakly positive on day 10 after osteoblast grafts, and nNOS expression increased a little at days 14-21, while it had no obvious peak value, and was weakly positive during the whole process. was a little weaker with a low and was found in the control group. However, iNOS and eNOS expression flat peak, and no obvious peak of nNOS CONCLUSION： Expression of iNOS and eNOS is time dependent, and its cell-orientation and value are different, which indicates that release of iN-OS and eNOS may regulate the process of osteoporotic fracture healing. Expression of nNOS is not significantly involved in the healing of osteoporotic fracture in the elderly.
出处
《中国临床康复》
CSCD
北大核心
2005年第38期1-3,共3页
Chinese Journal of Clinical Rehabilitation
基金
广东省重点科技攻关项目资助(9622050-03)~~
