摘要
为了探讨凝血酶通过损伤肾小球内皮细胞（ＧＥＣ）介导肾小球疾病的机制，观察了纯化的α凝血酶对体外ＧＥＣ增殖、生长状况以及表达纤溶酶原激活物（ＰＡ）、纤维连接蛋白（ＦＮ）的影响，并用水蛭素和抑肽酶进行了阻滞试验。采用噻唑蓝（ＭＴＴ）法测细胞增殖，纤维蛋白平板法测总ＰＡ活性，发色底物法测组织型纤溶酶原激活物（ｔ－ＰＡ）活性，ＥＬＩＳＡ和间接免疫荧光法检测ＦＮ含量。结果表明：（１）０．４～３．２ＮＩＨＵ／ｍｌ凝血酶显著促进ＧＥＣ的增殖和代谢，Ｐ＜０．０５；（２）３．２ＮＩＨＵ／ｍｌ凝血酶刺激３２小时后可诱导ＧＥＣ脱壁，这种作用可被水蛭素和抑肽酶阻断；（３）凝血酶明显增高培养上清中ＰＡ及ｔ－ＰＡ活性，Ｐ＜０．０１；（４）凝血酶显著减少ＧＥＣ培养上清中的ＦＮ含量，Ｐ＜０．０１。同时，细胞外基质中的ＦＮ含量也减少，且可被抑肽酶和水蛭素阻断。结论：凝血酶可导致ＧＥＣ增殖、脱壁，后者可被抑肽酶阻断，因而很可能与细胞贴壁所必需的ＦＮ等细胞外基质被升高的ＰＡ活性过度降解有关。
To clarify the mechanisms of thrombin contributing to the progression of glomerular di seases by injuring glomerular endothelial cells (GECs), we studied the effects of thrombin on GECs in vitro. Cell proliferation was detected with MTT incorporation, total plasminogen activator (PA) and tissue type PA (t PA) activities were detected with fibrin plate and chromatogenic substrate methods and fibronectin was detected with ELISA as well as indirect immunofluore scence. 0.4 3.2 NIH U/ml thrombin promoted GEC proliferation significantly ( P <0.05). Thrombin promoted cell detachment, which can be inhibited by hirudin or aprotinin. Thrombin enhanced total PA and t PA activities of GECs significantly ( P < 0.01). Fibronectin in the supernatants of thrombin stimulated GECs decreased significantly ( P <0 01) and in the extracellular compartment also decreased. The decrease was inhibited by hirudin and aprotinin. In conclusion, thrombin can induce GEC proliferation and GEC detachment. The latter is pro bably related to PA mediated over degradation of extracellular matrices such as fibronectin, which are needed for cell attachment.
出处
《中华内科杂志》
CAS
CSCD
北大核心
1997年第9期599-602,共4页
Chinese Journal of Internal Medicine
基金
国家自然科学基金
军队"九五"医药卫生科研基金
