中文摘要：

目的：多种实验已获知弱强度的电流可促进成骨细胞的增殖、分化和钙盐分泌，进而加速骨伤愈合，加快骨组织代谢。利用表面增强拉曼光谱，可在单细胞水平观察电流刺激下成骨细胞质膜蛋白结构的变化。 方法：实验于2006-06／08在南开大学泰达应用物理学院完成。出生4d的昆明品系乳鼠6只，体质量约50g，将1×10^9L^-1密度的成骨细胞用2．5g／L胰酶消化下来，置入到5mmol／L葡萄糖溶液（D-Hanks配置）中。做拉曼光谱的时候取出100μL含有成骨细胞的D-Hanks溶液，使一些成骨细胞固定到CaF2载玻片上。然后应用表面增强拉曼光谱技术，采集0．4mA和1．3mA不同电流强度作用下成骨细胞的拉曼光谱，并与电磁场作用前采集的拉曼光谱做对比。 结果：①成骨细胞加电流前后，分别得到30个细胞的拉曼光谱，并利用软件ongin7．0算出拉曼光谱的平均相对强度。②根据电流作用前10个细胞的拉曼光谱的相对峰值的平均值和电流作用后10个细胞的拉曼光谱的相对峰值的平均值，得出成骨细胞在受电流作用后，拉曼光谱中11个强峰的相对强度有明显的下降，表明在电流的作用后，成骨细胞膜蛋白构象有很大的变化。③表征蛋白质C—N伸缩振动的谱线1132cm^-1的相对强度，在受两种电流作用后，分别增加了128．57％、57．14％，表明细胞质膜蛋白从有序状态转变为无序状态。 结论：成骨细胞质膜蛋白在经过电流作用后，细胞膜结构发生较大变化，电流对稳定蛋白质二级结构的氢键和盐键有明显的影响。并且预示拉曼光谱有做为一种潜在的研究骨组织疾病诊断工具的可能性。

英文摘要：

AIM： It is reported that pulsed electromagnetic fields with low frequency and amplitude could induce osteoblasts to proliferate, differentiate and deposit calcium salts in several different ways, which consequently, accelerates bone healing and metabolism. In this study, the plasma membrane protein structure changes of osteoblast under the stimulation of electric current was observed at single cell level using the surface-enhanced Raman spectroscopy. METHODS： The experiment was performed in the College of Physics, Nankai University from June to August 2006. The osteoblasts （1 ×10^9 L^-1） from six 4-day-old Kunming mice of 50 g were digested by 2.5 g/L pancreatin, and put into 5 mmol/L glucose solution （D-Hanks）. 100 μL D-Hanks solution containing osteoblasts were fixed on CaF2 carry sheet glass. The Raman spectra of osteoblasts at the electric current stimulation of 0.4 mA and 1.3 mA were gained by surface-enhanced Raman spectroscopy. The result was compared with the spectra gained before applying the currents. RESULTS： ①There were Raman spectra of 30 cells. The average intensity of spectra was calculated with software origin 7.0. ②According to the average value of peak of Raman spectra of 10 cells before and after current stimulation, the 11 main spectrum peaks declined obviously, indicating under the electric current the protein structure of osteoblast plasma membrane had changed a lot.③The spectrum peak 1 132 cm 1 which represented the oscillation of C-N had increased 128.57% and 57.14%, respectively after stimulated by the two different currents, which indicated that the ordedy state of membrane proteins had changed into the disordedy state. CONCLUSION： The structure of osteoblast membrane changes obviously when the cells are stimulated by electric current. The current has strong influence on hydrogen bonds and salt bonds that can stabilize the second structure of proteins. These results indicate that Raman spectroscopy could be used as a diagnostic tool for bone tissue disease.