Basic introduction Chinese name: biliverdin mbth: classification of biliverdin: a morphological feature of biliverdin: dark green flaky or columnar crystals. Location: Precursors in bile and jaundice urine: bilirubin biliverdin, bilirubin biliverdin, metabolic pathway, serum biliverdin and biliverdin. Bilirubin is also called dehydrobilirubin. Bile pigment. It exists in bile and jaundice urine. Dark green flaky or columnar crystals. It turns black at about 300℃, does not melt after decomposition, and has no melting point. Soluble in methanol, ether, chloroform, carbon disulfide, benzene, insoluble in water. It can be obtained by oxidation of bilirubin. Used in biochemical research. See figure 1 for the specific structure. Figure 1 bilirubin bilirubin is generally considered as the main terminal metabolite of heme and the main bile pigment of poultry. Chickens and turkeys are discharged into bile in the form of biliverdin sodium salt. Birds lack the biliverdin reductase necessary to reduce biliverdin to bilirubin, so the percentage of bilirubin in total bile pigment of birds is very low. There is little or no bilirubin or biliverdin in serum or plasma of normal birds. Studies have found that the concentrations of bilirubin and biliverdin in chicken gallbladder are almost the same, but bilirubin is 75% more than biliverdin in bile of hepatobiliary duct. There was no hyperbilirubinemia when the bile duct was ligated on one side of the liver, but there was mild hyperbilirubinemia. When bilateral bile ducts are ligated, there will be trace biliverdinemia and obvious hyperbilirubinemia. Bilirubin in bird bile is very similar to that in dogs. This study shows that for chickens, biliverdin formed outside the liver or in the intrahepatic bile duct is reduced to bilirubin in the extrahepatic biliary system. The increase of serum bilirubin in carnivorous birds can occur in biliary obstruction or intravascular hemolytic diseases, while jaundice in parrots is extremely rare in clinic. Therefore, it is of little significance to use serum bilirubin to detect parrot's liver disease. The serum of parrots with liver disease or hunger is often green and contains urate, which is usually caused by hypercholesterolemia and hypercholesterolemia. Occasionally, parrots with chronic liver disease will develop jaundice, which may be caused by the nonspecific reduction of biliverdin to bilirubin. If the serum of normal birds is yellow, it is considered that this has nothing to do with bile red, but is caused by carrot pigment in food. Heme oxygenase (HO) is the rate-limiting enzyme of heme degradation, which can convert heme into biliverdin, CO and iron, and biliverdin can be reduced to bilirubin. It is known that there are three isoenzymes of HO, namely HO- 1, HO-2 and HO-3. HO-2 and HO-3 are constitutively expressed in large quantities, which may play their respective roles in heme binding in normal cells. HO- 1 is inducible and widely distributed in various tissues and cells of mammals. HO- 1 can be induced by many factors, such as oxidative stress, heat shock, ultraviolet radiation, ischemia reperfusion, heavy metals, bacterial lipopolysaccharide, cytokines, NO and its substrate heme. Recent studies have focused on the biological effects of enzymatic reaction products (biliverdin, cobalt and iron), which have antioxidant, anti-inflammatory, anti-apoptosis, signal transduction and immunomodulation, and inhibit the expression of adhesion molecules. The strong adaptive response of HO- 1 to various * * * indicates that HO-1may play an important role in preventing inflammatory reaction and oxidative tissue damage. Bilirubin is one of the products of the enzymatic reaction of HO- 1, and it is a bile pigment of triclosan. It is a dark green pigment and a precursor of bilirubin. It only exists in the bile of birds, amphibians or herbivores, but not in the bile and serum of normal people. In recent years, there have been many studies on the correlation between biliverdin and HO- 1 in animal experiments of other diseases, but the results are different. Some studies have found that biliverdin can react with HO- 1 and inhibit the expression of HO- 1 through negative feedback regulation, but others have found that it has no obvious effect on the expression of HO- 1. A large number of studies have confirmed that HO- 1 has neuroprotective effects on cerebral ischemia-reperfusion injury, such as antioxidation, anti-inflammation and anti-apoptosis. At the same time, more and more animal experiments show that biliverdin has a strong anti-inflammatory effect, which can not only inhibit the inflammatory reaction induced by lipopolysaccharide and protect acute lung injury in rats, but also inhibit the deterioration and development of adenotoxemia by reducing the production and release of inflammatory mediators, and also inhibit the expression of Toll-like receptor -4 and inflammatory factors in macrophages. In addition, exogenous administration of biliverdin can down-regulate the expression of adhesion molecules and inhibit the aggregation of white blood cells, thereby reducing the production of cytokines and chemokines or inhibiting the production of pro-inflammatory proteins such as cyclooxygenase -2 and cytochrome P450, and finally inhibiting the inflammatory reaction. In recent years, there are more and more reports on the application of biliverdin sheath in the protection of vascular injury, organ transplantation and other diseases, and its mechanism is mainly related to anti-inflammatory effect. It can play an anti-inflammatory role in the occurrence and development of many diseases, and slow down or improve the deterioration or prognosis of diseases by inhibiting inflammatory reaction, but its protective research on cerebral ischemia-reperfusion injury is less reported. Serum biliverdin is an intermediate product of hemoglobin and bilirubin. Lemberg said that complete liver function and adequate nutrition are necessary conditions for biliverdin to be converted into bilirubin. Bilirubin can also be oxidized by bilirubin, especially after liver depression and biliary tract. Bilirubin appears in the blood at this time, which may be caused by reflux. Bilirubin turns the skin green. When biliverdin is contained, intradermal injection of a small amount of 0. 1% potassium ferrocyanide solution can cause blue-green color. Bilirubin does not show Vandenberg reaction, but is determined by a special spectrophotometer. Usually serum bilirubin and biliverdin are parallel, but hemolytic jaundice has no biliverdin. Complete extrahepatic biliary obstruction, especially high biliverdin concentration, green skin. The improvement of nutritional status seems to reduce the serum biliverdin concentration. The production process of converting biliverdin into bilirubin The main product of bilirubin oxidation is biliverdin, and the yield of bilirubin can be improved by adjusting the process to convert it into bilirubin again. Based on fresh pig bile, the yield of bilirubin extracted by this method can reach 0.045% 1, operation method (1) Preparation of bile calcium Fresh pig bile was taken, filtered, and saturated limewater was added while stirring until the pH was =13 ~14. Then rapidly heating, when the temperature reaches about 40 DEG C, removing impurities such as foam, lipid and the like, keeping the temperature at 95 DEG C for 65,438+/-0.5 minutes without boiling, and filtering to obtain calcium cholate. The filtrate was used to recover crude pig bile acid, and bile calcium salt was acidified immediately. The requirements for saturated limewater are the same as the above methods. (2) Acidification: adding distilled water with the time of 10 to the calcium salt, then adding 2.0% NaHSO 3, and grinding into paste. When the temperature is lower than 20℃, dropwise add 1: 1 hydrochloric acid solution (hydrochloric acid: ice = 1: 1) while stirring until the pH value is 1 ~ 1.5. In the process of acidification, the speed of adding acid can be slightly faster at first, and then gradually slow down. If the bottom of acidification is not laid, it is easy to form lumps, so attention should be paid to prevention. As acidification approaches the end point, it becomes more and more concentrated. In order to prevent incomplete acidification, it can be ground and sieved, and then continue acidification. (3) Ethanol treatment At room temperature, add 10 times of ethanol to the acidified bile pigment, stir evenly, and adjust the pH to 3. Add 0.5% nahso _ 3, mix well, and stratify at low temperature in the dark. Under the same conditions, treat once more and draw thousands. Pay attention to the viscous bile pigment to grind and disperse evenly, so that ethanol can be fully contacted. The pH value should not be higher than 4. (4) Rinse with water. Put the bile pigment into hot water at 80℃ which is 0/0 times of/kloc-0, stir it evenly, keep the temperature at 10 minute, collect the floating bilirubin, and dehydrate it with ethanol. (5) extract dehydrated bile pigment with chloroform, add 0.5% NaHSO 3, and reflux extract with chloroform at 60℃ until the extract is colorless (less chloroform is used, and repeated extraction is effective). (6) distilling to recover chloroform, collecting at least a certain amount of distilled chloroform extract of refined bilirubin under reduced pressure, cooling to room temperature, adding 1%NaHSO 3 and anhydrous ethanol, adjusting pH to acidity, and standing in the dark at 4℃ for more than 12 hours to crystallize bilirubin. Collecting crystallized bilirubin, washing with hot distilled water, washing with ether, and vacuum drying to obtain refined bilirubin. 2, biliverdin into bilirubin conditions. (1) antioxidant should be added, (2) the pH is acidic, (8) it should be stored in the dark at low temperature 12 hours or more. Under the above three conditions, a part of biliverdin can be converted into bilirubin again.