正常和肝损害模型ASGP受体实验研究

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目的 通过对肝细胞去唾液酸糖蛋白受体 (ASGPR)功能的体内外分析来区别正常肼脏与慢性肝脏损害。方法 采用吸入CCl4 法建立慢性肝损害大鼠模型 ,用流式细胞仪 (FCM)技术异硫氰酸荧光素 新半乳糖人血清白蛋白 (FITC NGA)进行体外受体分析。SnCl2 还原法制备99Tcm NGA ,进行小鼠体内分布和正常及慢性肝损害模型大鼠ASGP受体显像研究。结果 FITC NGA结合的荧光道数值随肝损害程度的增加而降低。99Tcm NGA能迅速被肝脏摄取 ,其它脏器摄取均较低 ,肠道放射性随时间增加而增加。99Tcm NGA受体动态显像结果表明正常大鼠血本底清除快 ,注射 5min后肝脏显影清晰 ,而肝损害模型大鼠血本底清除较慢 ,注射 5min后 ,心、肺影仍存在 ,肝脏轮廓不清晰。简单动态分析显示 ,正常及肝损害模型大鼠心脏及肝脏的时间 放射性曲线差别较明显。“清除指数”HH15分别为 0 6 75± 0 10 6和 0 6 96± 0 10 3:“受体指数”LHL15分别为 0 980± 0 0 10和 0 949±0 0 2 5。结论 体外FCM分析可从细胞水平反映正常及损伤肝细胞ASGPR数量的变化。99Tcm NGA肝受体动态显像可反映肝细胞受体功能 ,而LHL15可较好反映肝细胞受体数量。 Objective To distinguish between normal hydrazine and chronic liver damage by in vitro and in vivo analyzes of hepatocyte asialoglycoprotein receptor (ASGPR) function. Methods Rat model of chronic liver injury was established by inhalation of CCl4 and in vitro receptor analysis was performed by flow cytometry (FITC NGA) using fluorescein isothiocyanate (FITC). The 99Tcm NGA was prepared by SnCl2 reduction method, and the ASGP receptor imaging in vivo was performed in vivo and in rats with normal and chronic liver injury. Results The number of FITC NGA-conjugated fluorescent channels decreased with the degree of liver damage. 99Tcm NGA can be quickly uptake by the liver, lower uptake by other organs, and increased intestinal radioactivity over time. The results of 99Tcm NGA receptor dynamic imaging showed that the blood clearance in normal rats was fast, the liver developed clearly 5 min after injection, while the blood loss in rats with liver damage was slow. After 5 min injection, heart and lung shadow still existed, Clear. A simple dynamic analysis showed that there was a significant difference in the time radioactivity curve of the heart and liver between normal and hepatic impairment rats. The “clearance index” HH15 was 0 6 75 ± 0 10 6 and 0 6 96 ± 0 10 3, respectively: “receptor index” LHL 15 was 0 980 ± 0 0 10 and 0 949 ± 0 0 2 5, respectively. Conclusion In vitro FCM analysis can reflect the changes of the number of ASGPR in normal and injured hepatocytes at the cellular level. Dynamic imaging of 99Tcm NGA hepatic receptor can reflect the function of hepatocyte receptor, while LHL15 can better reflect the number of hepatocyte receptors.
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