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Partial purification and characterization of hepatocyte growth factor from serum of hepatectomized rats
Abstract:
When rat serum was subjected to gel filtration on a Sephadex G-200 column, a factor, "hepatotropin", that promoted hepatocyte growth in primary culture was separated. Its Mr was about 150 KD and it was an anionic protein that was unstable on acid- and heat-treatments. Hepatotropin was purified 20-fold further by affinity chromatography on heparin-Sepharose CL-6B. The purified hepatotropin was effective at 20 micrograms/ml and maximally effective at 120 micrograms/ml, and its effect was additive with that of insulin plus epidermal growth factor. In rats after partial hepatectomy, the hepatotropin activity in the serum increased time-dependently reaching a maximum of about 5-fold the initial level 24 h after the operation. Various known growth factors, such as fibroblast growth factor, platelet derived growth factor, somatomedin, thrombin and transferrin, did not stimulate DNA synthesis in cultured hepatocytes. These results suggest that hepatotropin is a new growth factor.
Insights
Researchers discovered a new growth factor, hepatotropin, in rat serum that promotes liver cell growth. Its activity increases after liver surgery, suggesting a role in liver regeneration.
Area of Science:
- Biochemistry
- Cell Biology
- Hepatology
Background:
- Hepatocyte growth is crucial for liver regeneration.
- The specific factors regulating this process are not fully understood.
Purpose of the Study:
- To identify and characterize novel growth factors involved in hepatocyte proliferation.
- To investigate the role of a serum-derived factor in liver regeneration.
Main Methods:
- Gel filtration chromatography (Sephadex G-200) to isolate serum factors.
- Affinity chromatography (heparin-Sepharose CL-6B) for purification.
- Primary hepatocyte culture to assess growth-promoting activity.
- Partial hepatectomy in rats to study in vivo activity.
Main Results:
- A hepatocyte growth-promoting factor, termed hepatotropin, was isolated from rat serum.
- Hepatotropin is a ~150 kDa anionic protein, sensitive to acid and heat.
- Purified hepatotropin demonstrated dose-dependent growth promotion, additive with insulin and EGF.
- Serum hepatotropin levels significantly increased post-partial hepatectomy in rats.
Conclusions:
- Hepatotropin represents a novel growth factor distinct from known factors like FGF, PDGF, and IGF.
- This factor plays a significant role in stimulating hepatocyte growth and potentially liver regeneration.
