| Hepatocyte growth factor(HGF) is a kind of protein that can strongly stimulatehepatocyte caryocinetic, now we have learned it's amino acids sequence, andcloned humen cDNA which coding it as well as murine. HGF is a multy-origincytokine,was find in many organ such as liver, kidney and lunch. It was also findin many cell type. HGF is nultifunctional cytokine, possessing a lot of biologicalactivities, and performming function under physiological and pathologicalconditions. People have studied deeply on HGF and discovered it was involvedin maintaining liver, lung and kidney cells, furthermore, was involved in theorgans' regeneration and injury repair. It also has the ability of anti-fibrosis ofthe mentioned organs. Furthermore, HGF have significant curative effect onchronic hepatitis, acute hepatitis, medine hepatitis, hepatic injury, hepaticcirrhosis, liver transplantion and liver regeneration after partial resection. Animalexperiment indicated that HGF have acceptable result on repairing CCl4 inducedhepatic injury.HGF has an important role in tumorous generation and development. On onehand it act as and agent against some tumors, on the other hand, it can facilitatetumor growth, migration and invasion through regulating the express of adhesionmolecules, induce neogenesis and participate ECM degenaration. The way HGFexert it's function is started by binding to it's receptor c-Met, the binding cause toTyr phosphorylation and subsequence activate PTK activity, then initiationoutstanding proliferation, differentiation, cell movement adhesion and suchbiological events, promote hepatic cell, endothelial cell, endotheliocyte,melanocyte and hematopoietic cell growth, migration and morphogenesis. Undermany pathological conditions HGF upregulate it's express, such as in liver cancer,hepaticcirrhosis, HBP, pulmonary bibrosis injury of the lungs. It could be takenas detection index of such disease. In general, HGF has a wide prospect inchinical application as a multifunction cytokine.We performed the following work during the experiment we have used manypurification methods, afer serious summarize and comparison we finded out a setof purification method suitable to HGF.1. Dislysis cattle hepatocyte disruption containning HGF against water toremove salt, peptide and other micromolecule2. Cation-exchange chromatography. Potassium hydrogen phosphate buffer wasused, PH=7.4, with gradient elution, when the concentration of KCl 0.2M, wedetected HGF in the elutriant.3. Copper affinity chromatograph. Imidazole was used as eluant and HGF wasdetectable in elutriant after loading on 0.01Miminazole SDS-PAGE resultshow the purity of HGF is satisfacting. Dialysing removes potassiumhydrogen phosphate, KCl and iminazole then freeze drying, and measuringthe concentration of all the concentrate protein samples.We porformed MTT experiment, comparing the effect on cell proliferation of allsample and HGF's effect under different concentration. The results indicate HGFinduce cell proliferation strongly. At the same time we performed wound healingexperiments to measure the cell migration, and the results confirmed it canpromote cell migration. |
