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Preparation method of hepatocyte growth-promoting factors employing ultrasonic treatment

A technology of hepatocyte auxin and ultrasonic treatment, which is applied in the field of preparation of hepatocyte auxin by ultrasonic treatment, can solve the problems of large protein molecules, low safety, and low activity, and achieve high activity, high safety, and extraction high rate effect

Inactive Publication Date: 2015-03-04
HEFEI PINGGUANG PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing preparation methods have low yield, low activity, large protein molecules, and low safety.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] A kind of preparation method of ultrasonic treatment hepatocyte growth-stimulating hormone proposed by the present invention comprises the following steps:

[0033] S1. Pretreatment of suckling pig liver: take suckling pig liver, remove gallbladder, fascia and connective tissue, wash with water, drain and then wash with water for injection, then cut into small pieces or thin strips with a width of 3-5cm, and then Put it in a meat grinder and grind it into suckling pig liver particles with a particle size of 1-3mm;

[0034] S2. Homogenization: Add suckling pig liver granules obtained in S1 into a colloid mill and grind at a speed of 3100rpm for 2.5min to obtain suckling pig liver mince, and add suckling pig liver mince and water for injection into a blender at a weight ratio of 1:1, and Stir at a rotating speed of 310rpm for 14min to obtain suckling pig liver homogenate;

[0035] S3. Ultrasonic treatment: place the suckling pig liver homogenate obtained in S2 in an ultr...

Embodiment 2

[0042] A kind of preparation method of ultrasonic treatment hepatocyte growth-stimulating hormone proposed by the present invention comprises the following steps:

[0043] S1. Pretreatment of suckling pig liver: take suckling pig liver, remove gallbladder, fascia and connective tissue, wash with water, drain and then wash with water for injection, then cut into small pieces or thin strips with a width of 3-5cm, and then Put it in a meat grinder and grind it into suckling pig liver particles with a particle size of 1-3mm;

[0044] S2. Homogenization: add suckling pig liver granules obtained in S1 into a colloid mill and grind at a speed of 3000rpm for 3 minutes to obtain suckling pig liver mince, add suckling pig liver mince and water for injection into a blender at a weight ratio of 7:9, and grind at a speed of 300rpm Stir at a rotating speed of 16min to obtain suckling pig liver homogenate;

[0045] S3. Ultrasonic treatment: place the suckling pig liver homogenate obtained i...

Embodiment 3

[0052] A kind of preparation method of ultrasonic treatment hepatocyte growth-stimulating hormone proposed by the present invention comprises the following steps:

[0053] S1. Pretreatment of suckling pig liver: take suckling pig liver, remove gallbladder, fascia and connective tissue, wash with water, drain and then wash with water for injection, then cut into small pieces or thin strips with a width of 3-5cm, and then Put it in a meat grinder and grind it into suckling pig liver particles with a particle size of 1-3mm;

[0054] S2. Homogenization: Add suckling pig liver granules obtained in S1 into a colloid mill and grind at a speed of 3150rpm for 2.3 minutes to obtain suckling pig liver mince, and add suckling pig liver mince and water for injection into a blender at a weight ratio of 8:8.5, and Stir at a rotating speed of 305rpm for 15min to obtain suckling pig liver homogenate;

[0055] S3. Ultrasonic treatment: place the suckling pig liver homogenate obtained in S2 in ...

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Abstract

The invention discloses a preparation method of hepatocyte growth-promoting factors employing ultrasonic treatment. The method comprises the following steps: 1) pretreatment of porcine hepatocytes; 2) homogenizing; 3) ultrasonic treatment, namely processing porcine hepatocytes homogenate in an ultrasonic treatment machine in an ice bath by virtue of an ultrasonic wave to obtain ultrasonic homogenate; 4) enzymolysis, putting the ultrasonic homogenate into a reaction container, dropwise adding a hydrochloric acid until the pH value is 2.5-3.5, adding pepsase and stirring, and then putting the reaction container and contents of the reaction container in a water bath kettle to hydrolyze to obtain an enzymolysis material; 5) inactivation; 6) centrifuging; 7) coarse filtration, namely filtering a neutral centrifugal liquid by virtue of a hollow fiber ultrafiltration device of which the molecular weight cut off is 100KD and a hollow fiber ultrafiltration device of which the molecular weight cut off is 10KD to obtain coarse filtrate; and 8) ultrafiltration, namely carrying out ultrafiltration on the coarse filtrate by virtue of an ultrafiltration membrane of which the molecular weight cut off is 6,000D, so as to obtain a hepatocyte growth-promoting factor liquid. The method disclosed by the invention is simple and mild in process, and high in extraction rate; and the final product is relatively high in activity and high in safety.

Description

technical field [0001] The invention relates to the technical field of hepatocyte growth-stimulating hormone, in particular to a method for preparing hepatocyte growth-stimulating hormone by ultrasonic treatment. Background technique [0002] Hepatocyte growth-stimulating hormone is a drug that stimulates the DNA synthesis of newborn human and animal liver cells, can promote liver cell regeneration, accelerate the repair of liver tissue, restore liver function, improve the phagocytosis of liver Kupffer cells, and prevent intestinal The toxins in the liver can damage the liver cells, promote the repair of liver necrosis, and have a protective effect on liver cell damage, and can obviously improve the survival of liver failure. [0003] Hepatocyte growth-stimulating hormone is currently on the market, and it is prepared from fresh liver organs of dairy cows or suckling pigs through biochemical processing. However, the existing preparation method has low yield, low activity, l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K1/34
Inventor 王康林马婕苟希霞
Owner HEFEI PINGGUANG PHARMA
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