| 50%～75% liver disease patients have the active viral duplication and the liver inflammation change, approximately 40 million patients will appear the varied degree of liver harm, the liver cirrhosis and the liver function failure. In the whole world, 20 million people dead of this disease. The home position liver transplant was the only effective treatment to this disease up to now. But only few patients were able to obtain the liver to transplant because short of liver resource. How to control acute liver damage earlier, delay and prevent the occurrence and development of the last stage of liver disease, and how to develop the new drug which can cure the liver disease, is the goal of the domestic and foreign scholars.The liver has very strong regeneration function, the liver regeneration adjustment completes in the many kinds of cells factor and under other factor synergism regulation, including hepatocyte growth factor, insulin-like growth factor, epidermal growth factor, transforming growth factor and so on. As early as in 1964, after observed the animal liver partial excision, there maybe some kinds of body fluid factor induce the liver regeneration. Among them, HGF is one kind of powerful factor that can promote the fission, not only stimulates the synthesis of deoxyribonucleic acid (DNA) in the liver cell, but also stimulates its epithelial cell, the endothelial cell, the melanocyte cell and the corpuscle multiplication. In the kidney, HGF mainly produced in the kidney pellet area by the endothelial cell and the department membrane cell, in renal tubule area mainly expression to interstitial substance cell, endothelial cell and macrophage. The research of HGF mainly aims to the liver, when the liver excision and manyother factors like infections, to be poisoned, lack of the blood and so on cause the liver cell damaged, the density of plasma HGF and the c-met increased (take the liver excision patient for example, the density of plasma HGF can elevate 15～25 times, thus urges the liver cell multiplication and the differentiation, but the c-met expression restores to normal in the short time). HGF is a multi effect growth factor, it play an important role in the growth of the embryo and the organ, and it can participate in the repair of the organization through promote cell proliferation, it is product through interstitial cell, and is the strongest mitogen so far as now. It can promote the migration of cell, induce and start the liver regeneration, urge the morphogenesis function of organization organs, and it also can suppress the growth of certain liver cancer cells, it play an important role in repairing the damage of the liver, lung, and kidney. Its mainly function includes:①It can obviously stimulate the synthesis of the DNA of new born liver cell, promotes the restore of damage liver cell line grain and the rough endosarc net, promotes the regeneration of liver cell, accelerates the repair of the liver organization, restores the function of liver.②It can improve the swallow function of liver dry cell, prevent the damage to liver cell from the intestinal tract toxin, suppress the action of tumor necrosis factor(TNF) and Na+-K+-ATP enzyme, so it can promote liver necrosis repair. Simultaneously, it can reduce the activity of aminotransperase, serum bilirubin and proserozym.③It can protect the damage of liver cell by the carbon tetrachloride.④It can elevate the longevity of the liver failure that induced by the D- amino galactose. It is used to treat the heavy sickness hepatitis(acute, subac liver necrosis), the liver cirrhosis, the chronic activity hepatitis, heavy sickness acute icterohepatitis, drug poisoning, the allergy, and the radiotherapy causes all kinds of the liver damageand liver failure caused by the radiotherapy. In the recent 20 years, the domestic and foreign scholars have obtained a better curative effect in the study of using the cell factor to promote the liver regeneration to treat all kinds of liver damage.At present, our country develops varies drug to cure liver disease. The regeneration process is a quite complex process and it relates with many cell factors and physiological mechanism. the domestic research and development production pressed the liver cell growth factor to have the various biological effect, might obviously reduce the heavy sickness hepatitis and the case of fatality rate, and achieved the same level with embryo liver cell treatment group. Regarding the chronic hepatitis, HGF is better than the chest gland peptide group in reducing aminotransferase, serum bilirubin, improving the symptom aspect. According to the investigation, our country has product the liver cell growth factor for 10 years, the mainly molecular weight is a liver cell growth injection which composed by three protein that the molecular weight are 21.0KDa±10%, 31.5KDa±10% and 38.5KDa±10% and the liver growth hormone injection with molecular weight 10,000 below polypeptides compositions. The raw material mostly was new born or the mammalian, just like young mouse, suckling pig, young sheep, cow's liver. Hepatocyte growth-promoting factors injection is not only one active constituent, among them, two protein which molecular weight exceed 30 thousand account to 50%, but these protein are useless, they reduced the effective drug consumption to 50%, and these protein maybe induce the allergy problem. Hence, to remove these protein is very important to avoid these problems.This experimental selected fresh healthy suckling pig liver, and use the membraneemulsification to extract, after centrifugalization and filter with the filter diaphragm MW40KDa and 10KDa, we use the ion chromatography techniques and sieve chromatography to purify the product, and we get the 21KDa liver cell factors through SDS-PAGE separation, we named it liver regeneration augmenter preparation. Under certain condition, the protein molecular weight of protein's logarithm is negative related with the electrophoretic mobility. Therefore,we should pay attention to SDS and the protein union in the SDS-PAGE process (namely:1.4gSDS/g protein), if the proportion is improper, the accurate data cannot obtained. When use the SDS-polyacrylamide gelatin electricity to determine the molecular weight, must simultaneously make the standard curve is necessary.
|
PDF Full Text Request