Use of trypsin

A technology for trypsin and uses, applied in the field of trypsin, can solve problems such as toxic effects, destruction of cell components, etc.

Inactive Publication Date: 2014-06-11
HENAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, ROS are toxic to cells

Method used

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  • Use of trypsin
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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] In this example, the application of trypsin in scavenging free radicals.

[0026] 1. Materials and methods

[0027] 1. Materials

[0028] Escherichia coli wild-type strain MG1655 was donated by Professor Imlay of Illinois State University. Streak inoculate the dry powder of the above-mentioned strains stored in freeze-dry on LB plates, culture at 37°C for more than 16 hours, pick a single colony into the LB liquid medium, and cultivate at 37°C until the concentration of the bacterial solution is about 10 9 cells / ml, glycerol -20 ℃ to save the strains, for later use.

[0029] 2. Trypsin treatment

[0030] Trypsin (500U / mg) was purchased from Amersco, take 100mg.ml -1 Trypsin was added to O 2 ·- Incubate the resulting mixed system at 37°C for 30min, then add 25μl of 10mg.ml -1 Soybean trypsin inhibitor terminates the reaction.

[0031] 3. Sample processing

[0032] Sample with 0.5mM Cu 2+ Either incubate with 25mM EDTA for 0.5h or incubate with 1mM DDC at 28°C for...

Embodiment 2

[0053] The application of trypsin in anti-aging (delaying aging) in this example.

[0054] The tumor mice with no significant difference in the number of voluntary activities were selected and divided into two groups, 10 in each group. The control group was intragastrically given 10 mL / kg of normal saline according to the weight of the mice, and the experimental group was given intravenous injection of normal saline (10 mL / kg) containing 0.6 mg / mL trypsin according to the weight of the mice, and the survival time of the mice was recorded. The results showed that the mice in the control group died within 1 week, and the survival period of the mice in the trypsin treatment group was extended to 8 weeks.

Embodiment 3

[0056] In this example, the application of trypsin in anti-tumor (inhibition of tumor cell proliferation activity).

[0057] Hela cells in the logarithmic growth phase were collected, and the experimental group was treated with trypsin (0.6 mg / ml), and the control group was added with 200 μL of bovine serum culture medium, and the temperature was 5% CO at 37°C. 2 and fully saturated humidity CO 2 Cultivate in the incubator for 24-48 hours, and detect the cell survival rate by MTT method. The results showed a 30 percent decrease in tumor cell survival in the trypsin-treated group.

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Abstract

The invention discloses a use of trypsin, which belongs to the technical field of biological medicine. As shown in detection of O2.- concentration in an in-vivo and in-vitro superoxide anion generation system (for example, a VB2 reaction system) of an organism, the trypsin has significant scavenging activity to O2<.->, and a certain amount of H2O2 is generated in the system. Wherein the trypsin concentration and the initial concentration of the O2<.-> influence the scavenging velocity of O2<.-> and the generation rate of H2O2 (namely, the reaction rate of the trypsin scavenging free radicals), Cu<2+> is a key factor of O2<.-> scavenging activity in a trypsin reduction system. Since the trypsin has strong scavenging action on superoxide anion, H2O2 generated in the free radical scavenging reaction can inhibit the breeding activity of tumor cells and prolong the survival time of tumor mice, and the trypsin can be used for preparing anti-aging drugs, antineoplastic drugs and health care products.

Description

technical field [0001] The invention specifically relates to a use of trypsin, which belongs to the technical field of biomedicine. Background technique [0002] With the development of free radical medicine, it is recognized that many diseases are related to free radicals (such as cancer, aging, etc.). In the process of continuous contact with the outside world, factors such as respiration (oxidation reaction), pollution, and radiation exposure all affect the production of free radicals in the human body. For example, the body produces superoxide anion free radicals during the metabolic process. If the excess superoxide anion free radicals are not removed in time, they will accumulate in the body to produce cytotoxicity, destroy the structure and function of the biofilm, and cause various diseases. [0003] Reactive oxygen species (ROS) including superoxide anion (O 2 ·- ), hydrogen peroxide (H 2 o 2 ), hydroxyl radicals (·OH) and singlet oxygen (1O 2 ), etc., are the...

Claims

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Application Information

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IPC IPC(8): A61K38/48A23L1/29A61K8/66A61P39/06A61P35/00A61Q19/08A23L33/00
Inventor 李欣庞新跃王利平杨丽鹏李鑫玲尤晓颜乔家驹乔苏瑜
Owner HENAN UNIV OF SCI & TECH
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