Functions and applications of ubiquitin-specific protease 4 in treatment of fatty liver and type 2 diabetes

Abstract

Functions and applications of a USP4 gene in treatment of fatty liver and diabetes are disclosed. USP4 gene knockout mice and wild type C57 mice are adopted as experiment objects, a high-fat-diet induced obesity mouse model is utilized, and results show that compared with the wild type C57 mice, the USP4-KO mice show obesity, fasting blood glucose levels of the USP4-KO mice are higher than those of WT mice in a contrast group, and liver functions of the USP4-KO mice are obviously worse than those of the WT mice. Intraperitoneal injection glucose tolerance experiments show that glucose tolerance of the USP4-KO mice is obviously weakened. Liver weights, liver / weight ratios, liquid component results, and the like all prove that fatty liver lesion of the USP4-KO mice provided with high-fat diets is obviously serious and lipid accumulation is obviously increased, and therefore the USP4 can be adopted as a target for screening medicines treating the fatty liver and / or type 2 diabetes, and an accelerant of the USP4 can be used for preparing medicines treating the fatty liver and / or type 2 diabetes.

Description

technical field

[0001] The invention belongs to the field of gene function and application, and particularly relates to a ubiquitin-specific protease 4 (ubiquitin-specific protease 4, USP4) as a target gene in the preparation of a drug for preventing, alleviating and / or treating fatty liver and / or type II diabetes application in medicine. Background technique

[0002] With the change of people's lifestyle, especially the excessive intake of high-energy food and the reduction of physical activity, the incidence of diabetes and fatty liver has risen sharply, which has seriously endangered people's health. The increase in the incidence of type Ⅱ diabetes (Type2diabetes, T2DM) has led to a gradual increase in the prevalence and severity of nonalcoholic fatty liver disease. Non-alcoholic fatty liver disease (NAFLD) in non-obese people [body mass index (BMI) <25kg / m 2 】The incidence rate in 25kg / m is about 18.4%. 2 <BMI<30kg / m 2 About 63.4% of the population, and in t...

Examples

Embodiment 1

[0056] [Example 1] Fatty liver in mice and type Ⅱ diabetes model (diet induced obesity, DIO) were obtained

[0057] (1) Grouping of experimental animals: 8-week-old, male, WT mice and USP4-KO mice were selected and given two special diets, D12942 high-fat diet (High fat diet, HFD) and D12450B low-fat diet (Normal chow , NC) feeding, that is WTNC group, KO NC group, WT HFD group, KO HFD group a total of 4 groups.

[0058] (2) The model induces the operation process through high-fat feed:

[0059]WT and KO mice were used to establish DIO models, and phenotype correlation analysis was performed to clarify the role of USP4 gene on fatty liver and type Ⅱ diabetes. 8-week-old, male, WT mice and USP4-KO mice were selected and fed with two special diets, D12942 high-fat diet (Highfat diet, HFD) and D12450B low-fat diet (Normal chow, NC), respectively, WT NC group, KO NC group, WT HFD group, KO HFD group, a total of 4 groups. The food intake of the mice was recorded in detail every ...

Embodiment 2

[0060] [Example 2] Determination of mouse body weight, blood sugar level, and serum insulin level

[0061] (1) Detection of fasting body weight of mice

[0062] 1) Weight detection.

[0063] ①Fasting: fast the mice to be tested at 8:00 am (without water), and start the experimental operation at 2:00 pm.

[0064] ② Weighing: Weigh at 0 weeks, 2 weeks, 4 weeks, 8 weeks, 10 weeks, and 12 weeks respectively, put a small plastic bucket on a dynamic electronic balance, grab the mouse, and put it into the weighing bucket , measure body weight and record data. Feed amount detection: After the weighing operation is completed, add feed to the mice, and record the amount of feed for the mice on the dynamic electronic balance.

[0065] (2) Fasting blood glucose level detection experiment

[0066] All the mice to be tested were fasted from 8:00 am to 2:00 pm (without water), that is, the experimental operation was started after 6 hours of fasting.

[0067] ① Blood glucose meter prepar...

Embodiment 3

[0090] [Example 3] Glucose tolerance test (intraperitoneal glucose tolerance test, IPGTT)

[0091] On the 10th week of the experiment, the intraperitoneal injection of glucose test (IPGTT) was performed to evaluate the glucose tolerance of the mice.

[0092] (1) Before measuring blood glucose, measure the fasting body weight of the mice, and calculate the injection volume of glucose based on 10 μL / g.

[0093] (2) First detect the fasting blood glucose at 0 minutes before the glucose injection, and inject the glucose solution intraperitoneally quickly after the detection is completed.

[0094] (3) Operation method of intraperitoneal injection: ①Fix the mouse; grab the mouse, grab the tail of the mouse with the little finger and ring finger of the left hand, and grab the neck of the mouse with the other three fingers, so that the head of the mouse is downward, and the The abdomen of the mouse is fully exposed. ②Needle positioning and injection: insert the needle from the side ...

technical field

[0001] The invention belongs to the field of gene function and application, and particularly relates to a ubiquitin-specific protease 4 (ubiquitin-specific protease 4, USP4) as a target gene in the preparation of a drug for preventing, alleviating and / or treating fatty liver and / or type II diabetes application in medicine. Background technique

[0002] With the change of people's lifestyle, especially the excessive intake of high-energy food and the reduction of physical activity, the incidence of diabetes and fatty liver has risen sharply, which has seriously endangered people's health. The increase in the incidence of type Ⅱ diabetes (Type2diabetes, T2DM) has led to a gradual increase in the prevalence and severity of nonalcoholic fatty liver disease. Non-alcoholic fatty liver disease (NAFLD) in non-obese people [body mass index (BMI) <25kg / m 2 】The incidence rate in 25kg / m is about 18.4%. 2 <BMI<30kg / m 2 About 63.4% of the population, and in t...

Examples

Embodiment 1

[0056] [Example 1] Fatty liver in mice and type Ⅱ diabetes model (diet induced obesity, DIO) were obtained

[0057] (1) Grouping of experimental animals: 8-week-old, male, WT mice and USP4-KO mice were selected and given two special diets, D12942 high-fat diet (High fat diet, HFD) and D12450B low-fat diet (Normal chow , NC) feeding, that is WTNC group, KO NC group, WT HFD group, KO HFD group a total of 4 groups.

[0058] (2) The model induces the operation process through high-fat feed:

[0059]WT and KO mice were used to establish DIO models, and phenotype correlation analysis was performed to clarify the role of USP4 gene on fatty liver and type Ⅱ diabetes. 8-week-old, male, WT mice and USP4-KO mice were selected and fed with two special diets, D12942 high-fat diet (Highfat diet, HFD) and D12450B low-fat diet (Normal chow, NC), respectively, WT NC group, KO NC group, WT HFD group, KO HFD group, a total of 4 groups. The food intake of the mice was recorded in detail every ...

Embodiment 2

[0060] [Example 2] Determination of mouse body weight, blood sugar level, and serum insulin level

[0061] (1) Detection of fasting body weight of mice

[0062] 1) Weight detection.

[0063] ①Fasting: fast the mice to be tested at 8:00 am (without water), and start the experimental operation at 2:00 pm.

[0064] ② Weighing: Weigh at 0 weeks, 2 weeks, 4 weeks, 8 weeks, 10 weeks, and 12 weeks respectively, put a small plastic bucket on a dynamic electronic balance, grab the mouse, and put it into the weighing bucket , measure body weight and record data. Feed amount detection: After the weighing operation is completed, add feed to the mice, and record the amount of feed for the mice on the dynamic electronic balance.

[0065] (2) Fasting blood glucose level detection experiment

[0066] All the mice to be tested were fasted from 8:00 am to 2:00 pm (without water), that is, the experimental operation was started after 6 hours of fasting.

[0067] ① Blood glucose meter prepar...

Embodiment 3

[0090] [Example 3] Glucose tolerance test (intraperitoneal glucose tolerance test, IPGTT)

[0091] On the 10th week of the experiment, the intraperitoneal injection of glucose test (IPGTT) was performed to evaluate the glucose tolerance of the mice.

[0092] (1) Before measuring blood glucose, measure the fasting body weight of the mice, and calculate the injection volume of glucose based on 10 μL / g.

[0093] (2) First detect the fasting blood glucose at 0 minutes before the glucose injection, and inject the glucose solution intraperitoneally quickly after the detection is completed.

[0094] (3) Operation method of intraperitoneal injection: ①Fix the mouse; grab the mouse, grab the tail of the mouse with the little finger and ring finger of the left hand, and grab the neck of the mouse with the other three fingers, so that the head of the mouse is downward, and the The abdomen of the mouse is fully exposed. ②Needle positioning and injection: insert the needle from the side ...

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