Application of milk-derived polypeptide and chimeric peptide thereof in preparation of medicine for promoting adipocyte energy metabolism

A fat cell, energy metabolism technology, applied in the field of peptide and cell metabolism, can solve the problem that the content is difficult to meet the requirements of obesity treatment

Pending Publication Date: 2021-10-15
NANJING MATERNITY & CHILD HEALTH CARE HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the content and activity of brown fat reached its peak in the neonatal period. Although there is a certa

Method used

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  • Application of milk-derived polypeptide and chimeric peptide thereof in preparation of medicine for promoting adipocyte energy metabolism
  • Application of milk-derived polypeptide and chimeric peptide thereof in preparation of medicine for promoting adipocyte energy metabolism
  • Application of milk-derived polypeptide and chimeric peptide thereof in preparation of medicine for promoting adipocyte energy metabolism

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Unless otherwise specified, the peptide chains in this example were synthesized by Shanghai Keyept Biological Co., Ltd. with a purity of >98%. This embodiment provides an isolated milk-derived polypeptide, the amino acid sequence of which is VKEAMAPK, and the milk-derived polypeptide can promote energy metabolism of white adipocytes and brown adipocytes.

[0032] As an embodiment, the transmembrane sequence GRKKRRQRRR is connected to the N-terminus of the milk-derived polypeptide to obtain the chimeric polypeptide GRKKRRQRRRVKEAMAPK shown in SEQ ID NO.2; molecular formula C 93 h 175 N 41 o 22 S, average molecular weight 2.25 kDa, average isoelectric point pH 12.471.

[0033] As another embodiment, the transmembrane sequence CYGRKKRRQRRR is connected to the N-terminus of the milk-derived polypeptide to obtain the chimeric polypeptide CYGRKKRRQRRRVKEAMAPK shown in SEQ ID NO.3; molecular formula C 108 h 197 N 45 o 26 S, average molecular weight 2.52 kDa, average isoel...

Embodiment 2

[0038] Example 2 Seahorse detects the effect of chimeric polypeptide on the energy metabolism of adipocytes

[0039] 1. Experimental method

[0040] The human white and brown adipose precursor cells in the logarithmic growth phase were planted in Seahorse cell culture plates, cultured in DMEM / F12 medium until the cells covered the culture plates, and continued to fuse for 24 hours to induce cell differentiation. Among them, the white adipocytes were induced and cultured with DMEM medium containing 5 mg / L insulin, 0.5 mM IBMX, 1 mM dexamethasone, 1 μM rosiglitazone and 10% FBS as inducer I for 4 days, and then replaced with 5 mg The DMEM medium of / L insulin and 5% FBS was used as inducer II, and continued to induce cultured cells for 4 days; while the brown fat was induced by adding 1nM T3 (triiodothyronine) on the basis of the above inducer I, and then transformed into The culture was continued for two days for Inducer II.

[0041] On the 8th day of induced differentiation ...

Embodiment 3

[0044] Example 3 Western Blot detection of the effect of chimeric polypeptide on the expression of UCP1 protein in adipocytes

[0045] 1. Experimental method

[0046] Refer to the method of Example 2 to collect protein samples after treating the cells, use RIPA lysate to extract the total protein, and use the BCA method to quantify the above-mentioned protein. According to the calculated protein concentration, use 1x loading buffer (Loadingbuffer) to adjust the sample concentration To be consistent, convenient for later sample loading. 10% SDS-PAGE gel was used for vertical electrophoresis, and after electrophoresis, the separated protein was transferred to PVDF membrane by wet transfer method. After transmembrane transfer, incubate with UCP1 antibody (Abcam / ab155117) diluted 1:2000 at 4 degrees overnight, wash and incubate with anti-rabbit secondary antibody (diluted 1:5000) the next day, and finally use chemiluminescence to develop protein Bands.

[0047] 2. Experimental ...

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Abstract

The invention discloses an application of a milk-derived polypeptide and a chimeric peptide thereof in preparation of a medicine for promoting adipocyte energy metabolism, and relates to a milk-derived octapeptide and a chimeric polypeptide formed by connecting the milk-derived octapeptide and a cell-penetrating peptide. Through Seahorse detection and tests on the expression quantity of an important fat cell metabolism marker UCP1 protein and the expression quantity of PPARalpha, PGC1alpha and DIO2 genes, it is proved that a certain dose of chimeric polypeptide can significantly promote energy metabolism of white fat cells and brown fat cells; and the invention is of great significance to research on adipocyte metabolism and industrial implementation of biological agents for treating obesity.

Description

technical field [0001] The invention relates to the fields of polypeptide and cell metabolism, in particular to the application of a milk-derived polypeptide and its chimeric peptide in the preparation of drugs for promoting fat cell energy metabolism. Background technique [0002] Obesity is a metabolic disease caused by long-term energy intake exceeding consumption, and excess energy is stored in the body in the form of fat. Obese patients are often accompanied by serious complications such as type 2 diabetes, cardiovascular disease, hyperlipidemia, hypertension, and tumors, thus causing great harm. In the past 40 years, the global obesity population has increased from 105 million to 641 million, making obesity a serious global public health problem. my country's obesity problem is equally serious, and the total number of obese people (90 million) has ranked first in the world. It is foreseeable that the huge obese population will seriously aggravate my country's medical ...

Claims

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Application Information

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IPC IPC(8): C07K14/47C07K19/00A61K38/17A61K47/64A61P3/04
CPCC07K14/4732A61K47/64A61P3/04C07K2319/10A61K38/00
Inventor 崔县伟季晨博
Owner NANJING MATERNITY & CHILD HEALTH CARE HOSPITAL
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