蛋白酪氨酸激酶的ELISA测定方法及其在脾虚证研究中的应用

Description and Application of An Enzyme-Linked Immunosorbent Assay for the Detection of Protein Tyrosine Kinase Activity and the Study on Spleen-Deficiency Syndrome

目的:从细胞信号传导角度探讨脾虚证的免疫病理机制并为之建立相应的实验方法。方法:建立固相 ELISA 法,分别检测利血平脾虚模型小鼠及临床脾虚患者淋巴细胞蛋白酪氨酸激酶(PTK)活力。结果:新建的 ELISA 法简便可靠,敏感易行。应用该方法研究发现:脾虚模型小鼠脾淋巴细胞浆及膜性 PTK 活力均显著降低;脾虚患者外周血淋巴细胞胞浆PTK 活力同样明显低于健康人。结论:脾虚证淋巴细胞内 PTK 活力低下。

Objective:To explore the immunopathologic mechanism of Spleen Deficiency Syndrome(SDS)in the respect of cel- lular signal transduction and establishment of related experimental method.Methods:A solid-phase ELISA method was developed to detect the protein tyrosine kinase(PTK)activity in the lymphocytes of both clinical patients with SDS and reserpine-induced SDS mice.Results:The newly established ELISA was proved to be simple,dependable,and sensitive. Using this method,it was found that the PTK activities in the membrane and cytosolic extracts from splenic cells of reser- pine-induced SDS mice were significantly lower than that of normal.The PTK activity in the cytosol of peripheral blood lymphocytes of SDS patients were also reduced markedly.Conclusion:PTK activity was reduced in the lymphocytes of those individuals with SDS.