鼠肝星状细胞体内与体外激活的显微拉曼光谱

In vitro and in vivo Activation of Mouse Liver Astral Cell by Confocal Raman Microspectroscopy

采用密度梯度离心法从肝组织中分离、提纯肝星状细胞,进行常规细胞鉴定后,通过体外培养诱导肝星状细胞体外活化,在不同的时间点上进行原位拉曼光谱表征;通过一次性腹腔注射CCl4诱导鼠急性肝损伤,取不同的时间点的肝损伤组织做拉曼光谱表征,并以肝组织的光谱变化来间接反映肝星状细胞的体内活化.结果表明,用拉曼光谱能快速、灵敏地监测肝星状细胞体内和体外活化过程中的分子变化,可为肝纤维化的早期诊断提供依据.

The feasibility of a novel and efficient diagnostic method for liver fibrosis via Raman spectroscopy was studied. Confocal Raman Spectroscopy（CRS） was utilized to monitor the molecular changes of rat hepatic stellate cells（HSCs） in vitro as well as in vivo activation. In vitro activation was induced by growth in uncoated plastic plates while the in vivo activation was induced by a single intraperitoneal injection of carbon tetrachloride（ CCl4 ）. Raman spectroscopy in combination with biological method perfectly established a correlation between spectral changes and morphologic changes within HSCs. The biochemical changes of HSCs during the activation such as the loss of retinoid, the increase of a-helical protein, and the increased production of extracellular matrix proteins were observed by CRS. Raman spectroscopy combined with fiber optical probe could be potentially accomplished in vivo detection, which can lead to a novel and efficient diagnosis for liver fibrosis.