Application of macrophages in obesity treatment and blood sugar regulation

A macrophage and obesity technology, applied in the field of biomedicine, can solve the problems of difficult anti-inflammatory effects and short half-life, and achieve the effects of lowering blood sugar levels, lowering blood sugar, and reducing body weight

Pending Publication Date: 2021-08-13
鲲石生物科技(深圳)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, macrophages themselves do not secrete IL-4, and the half-life of IL-4 in the body is very short, the effective time is about 2-10 minutes,

Method used

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  • Application of macrophages in obesity treatment and blood sugar regulation
  • Application of macrophages in obesity treatment and blood sugar regulation
  • Application of macrophages in obesity treatment and blood sugar regulation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Preparation of a macrophage cell line stably expressing IL-4 protein (that is, RAW264.7 cells or IL-4-RAW cells infected with lentivirus pCDH-IL-4) and detection of its secreted IL-4 protein.

[0042] 1. Construction of recombinant plasmid pCDH-IL-4.

[0043]Insert the IL-4 gene between the recognition site and the BamHI recognition site of the restriction endonuclease EcoRI of the vector pCDH-MSCV-MCS-EF1α-copGFP-T2A-Puro (the nucleotide sequence is as shown in SEQ ID No. 1 Shown), the recombinant plasmid pCDH-IL-4 was obtained. Recombinant plasmid pCDH-IL-4 expresses IL-4 protein, and its amino acid sequence is shown in SEQ ID No. 2.

[0044] 2. Preparation of lentivirus pCDH-IL-4 concentrate.

[0045] 1. Inoculate the 293T cells grown to the logarithmic growth phase in a petri dish (60 mm in diameter) containing 10 mL of conventional medium, at 37 °C, 5% CO 2 to cultivate.

[0046] 2. Gently mix 1 μg pVSVg, 3 μg psPAX2, 4 μg recombinant plasmid pCDH-IL-...

Embodiment 2

[0067] Example 2 Survival of IL-4-RAW cells in mice.

[0068] The survival of IL-4-RAW cells infused into the peritoneal cavity of mice was analyzed by flow cytometry.

[0069] (1) Intraperitoneal injection of IL-4-RAW cells. IL-4-RAW cells in the exponential growth phase were digested with trypsin (Gibco), washed 3 times with PBS (Gibco), and then washed in PBS at 5×10 7 cell / mL concentration to make a single cell suspension. Then the mice were intraperitoneally injected with 1×10 7 cells, that is, each mouse was intraperitoneally injected with 200 µL of the above cell suspension.

[0070] (2) Flow cytometric detection of peritoneal cells. After completing step (1), at the indicated time points, mice were injected intraperitoneally with 5 mL of PBS, the abdomen was pressed slowly, and intraperitoneal fluid was collected 5 minutes later. After centrifugal filtration, the proportion of GFP-positive cells was detected by flow cytometry.

[0071] Experimental results such...

Embodiment 3

[0072] Example 3 Effect of IL-4-RAW cells on body weight of obese mice.

[0073] C57BL / 6 wild-type mice are products of Beijing Huafukang Biotechnology Co., Ltd.

[0074] PBS buffer is pH7.2-7.4, 67mM PO4 (Ca and Mg free) PBS buffer.

[0075] The experimental method is as follows.

[0076] Step 1. Take 20 C57BL / 6 wild-type mice of 6-8 weeks old, feed them with high-fat diet (Resarch Diets, 50% fat content), and measure their body weight every week.

[0077] Step 2. Three weeks after completing step 1, 20 C57BL / 6 wild-type mice were randomly divided into two groups (10 mice in each group) and a PBS group and an IL-4-RAW group. Then proceed as follows.

[0078] PBS group (negative control group): each mouse was intraperitoneally injected with 200 μl PBS buffer solution.

[0079] IL-4-RAW group (experimental group): Each mouse was injected with 200 μL of IL-4-RAW cell suspension into the tail vein.

[0080] The preparation method of IL-4-RAW cell suspension is as follows: co...

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Abstract

The invention belongs to the field of biomedicine, and particularly relates to application of macrophages expressing IL-4 protein in obesity treatment and blood sugar regulation. The application of the macrophage and a composition comprising the macrophage is as follows: (1) application in preparation of products for treating obesity; (2) application in preparation of products for reducing blood sugar; and (3) application in preparation of products for treating diabetes caused by obesity. The macrophage is a macrophage expressing an IL-4 (Interleukin-4) protein. When intraperitoneal injection is carried out on obese mice by using the prepared macrophage expressing the IL-4 protein, namely a lentivirus pCDH-IL-4 infected RAW264.7 cell, the weight of the mice can be effectively reduced, and the blood glucose level of the mice after glucose uptake is reduced. Therefore, it is proved that macrophages expressing the IL-4 protein can treat obesity and reduce blood sugar, new therapeutic drugs and methods are provided for clinically treating obesity and other diseases such as diabetes caused by obesity, and important application value is achieved for treating the obesity and other diseases such as the diabetes caused by obesity.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the new medical application of engineered macrophages, in particular to the application of macrophages expressing IL-4 protein in treating obesity and regulating blood sugar. Background technique [0002] With the rapid development of the economy and the acceleration of urbanization, people's lifestyle and dietary structure have undergone tremendous changes, and the overweight and obese population is increasing year by year. Studies at home and abroad have shown that obesity is an important factor in inducing hypertension, cardiovascular and cerebrovascular diseases, type 2 diabetes, hyperlipidemia, fatty liver and other diseases. A type of obesity in which fat deposition in the body begins to develop centered on the heart and abdomen is called central obesity or central obesity, which is a type of obesity that is more harmful than systemic obesity. The World Health Organization has list...

Claims

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Application Information

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IPC IPC(8): A61K35/15A61K38/20A61P3/04A61P3/10C12N15/867C12N15/24C12N5/10
CPCA61K35/15A61K38/2026A61P3/04A61P3/10C12N15/86C07K14/5406C12N5/0645C12N2740/15043C12N2800/107C12N2510/00
Inventor 尹秀山
Owner 鲲石生物科技(深圳)有限公司
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