链霉蛋白酶P、糖苷酶对低分子量肝细胞生长素生物活性的影响

Effect of pronase P and glycosidase digestions on bioactivity of low molecular weight hepatopoitin

目的研究链霉蛋白酶P、糖苷酶对低分子量肝细胞生长素生物活性的影响。方法用链霉蛋白酶P、N-糖苷酶F、O-糖苷酶分别对低分子量肝细胞生长素消化,观察消化后低分子量肝细胞生长素以及未经消化的低分子量肝细胞生长素和促肝细胞生长素对人肝癌细胞株QGY、HepG2、原代大鼠肝细胞的增殖作用,用3H-胸腺嘧啶核苷掺入法测定这些细胞的增殖情况。结果低分子量肝细胞生长素与促肝细胞生长素对人肝癌细胞株QGY、HepG2、原代大鼠肝细胞均有明显增殖作用(P<0.01);低分子量肝细胞生长素与促肝细胞生长素均对人肝癌细胞株QGY增殖作用最明显(P<0.01);低分子量肝细胞生长素增殖作用较促肝细胞生长素明显(P<0.01)。低分子量肝细胞生长素经链霉蛋白酶P、O-糖苷酶消化前后对人肝癌细胞株QGY、HepG2、原代大鼠肝细胞增殖作用无影响(P>0.05);低分子量肝细胞生长素经N-糖苷酶F消化后对人肝癌细胞株QGY、HepG2、原代大鼠肝细胞增殖作用明显减弱,差异有统计学意义(P<0.01)。结论低分子量肝细胞生长素促进人肝癌细胞株QGY、HepG2、原代大鼠肝细胞增殖作用比促肝细胞生长素更显著。

Objective To study the effect of pronase P and glycosidase on activity of low molecular weight hepatopoitin. Methods Hepatopoitin was disgested by pronase P, N-glycosidase F,O-glycosidase. QGY, HepG2 cells and primary rat hepatocytes were cultured separately with hepatopoitin ,hepatopoitin disgested by pronase P and glycosidase and PHGF. Cell proliferation was detec- ted by their 3H-TdR uptake. Results Hepatopoitin and PHGF could stimulate the proliferation of QGY,hepG2 cells and primary rat hepatocytes in vitro. The proliferation of QGY cells was the most significant in hepatopoitin the same as PHGF. The proliferation was not significantly different in the gepatopoitin disgested by pronase P,O-glycosidase（P〈0.05）. Hepatopoitin was resistant to enzyme digestion with pronase P,O-glycosidase, but would be inactived by N-glycosidase F digestion. Conclusion Low molecular weight hepatopoitin has more cell proliferation than PHGF in QGY, hepG2 cells and primary rat hepatocytes. Low molecular weight hepatopoitin is a glycopeptid containing N-sugar chains,which is essential for keeping its bioactivity.