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①目的用改良亲和层析法从人胎盘中分离肝细胞生长因子和肝细胞生长抑制物质。②方法取３０ｇ人胎盘加４倍体积（Ｗ／Ｖ）的缓冲溶液，匀浆，４℃，离心（１１０００ｒ／ｍｉｎ）３０ｍｉｎ，收集上清液；用４５％饱和度硫酸铵进行盐析。沉淀复溶后对洗脱缓冲溶液透析２０ｈ，再进行Ｓｅｐｈａｒｏｓｅ４Ｂ凝胶过滤柱层析。最后，将层析第２峰过肝素－ＳｅｐｈａｒｏｓｅＣＬ－６Ｂ亲和层析柱，用改良的洗脱方法进行亲和层析。③结果在分离肝细胞生长因子的同时，还分离得到一种肝细胞生长抑制物质。肝细胞生长因子可明显刺激原代培养的大鼠肝细胞ＤＮＡ的合成（ｔ＝６４８．３９，Ｐ＜０．０１），而肝细胞生长因子则抑制原代培养的大鼠肝细胞的ＤＮＡ合成（ｔ＝－１０．４４，Ｐ＜０．０１）。④结论改良亲和层析法在分离效率方面比传统方法有明显提高，而用该法分离到的肝细胞生长抑制物质可能参与了肝细胞的发育调节。
Objective To isolate hepatocyte growth factor and hepatocyte growth inhibitory substance from human placenta by modified affinity chromatography. Methods 30g human placenta plus 4 times volume (W / V) buffer solution was homogenized and centrifuged (11000r / min) at 4 ° C for 30min. The supernatant was collected and salted out with 45% saturated ammonium sulfate. The precipitate was reconstituted for elution buffer solution dialysis 20h, followed by Sepharose 4B gel filtration column chromatography. Finally, the second peak, Heparin-Sepharose CL-6B affinity chromatography, will be chromatographed and subjected to affinity chromatography using a modified elution method. ③ results in the separation of hepatocyte growth factor, but also isolated a hepatocyte growth inhibitory substance. Hepatocyte growth factor significantly stimulated DNA synthesis in primary cultured rat hepatocytes (t = 648.39, P <0.01), whereas hepatocyte growth factor inhibited DNA synthesis in primary cultured rat hepatocytes (T = -10.44, P <0.01). ④ Conclusion Improved affinity chromatography method in the separation efficiency was significantly improved than the traditional method, while using the method of hepatocyte growth inhibitory substances may be involved in the regulation of the development of liver cells.