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Silkworm polypeptide as well as preparation method and application thereof

A technology for silkworm chrysalis and diabetes, applied in the field of medicine

Inactive Publication Date: 2015-01-28
SUZHOU PULUODA BIOLOGICAL SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are no well-developed compounds extracted from silkworm chrysalis for the prevention or treatment of diabetes

Method used

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  • Silkworm polypeptide as well as preparation method and application thereof
  • Silkworm polypeptide as well as preparation method and application thereof
  • Silkworm polypeptide as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0011] Preparation method of polypeptide

[0012] Boil silkworm chrysalis in water for 2 hours, cool to room temperature; add 1% papain and 0.5% alkaline protease to hydrolyze for 4 hours; use macroporous resin to separate protein according to molecular weight, and then use semi-preparative RP-HPLC to separate and purify, and the obtained product is frozen After drying, it is silkworm chrysalis polypeptide 1. The silkworm chrysalis polypeptide 1 extracted by the above method is identified by mass spectrometry, and its sequence is RLGRRLSEDPATPADQ, which is a new sequence. You can also entrust Shanghai Sangong Synthetic Co., Ltd.

Embodiment 2

[0014] In Vitro Cell Viability Determination of Silkworm Chrysalis Polypeptide 1

[0015] The MTT colorimetric method was used. Insulinoma cells (INS-1 cells) with logarithmic growth were added to 96-well culture plate at 1.0×105, and cultured for 24 hours. Different concentrations of the experimental drug silkworm chrysalis polypeptide 1 were added to the experimental wells and positive drug control wells; the blank group was added same volume of solvent. Set up five duplicate wells in every well, culture 48h, add MTT in every hole, after 4h of action, add DMSO, hatch 30min, measure the absorbance A value at 570nm place of microplate reader, according to the formula cell growth proliferation rate=(experimental group absorbance value / Absorbance value of control group -1)×100%. The calculated EC50 of the experimental drug was 15.04nmol / L.

Embodiment 3

[0017] In vivo hypoglycemic experiment of silkworm chrysalis polypeptide 1

[0018] Kunming mice were fed with high-fat and high-sugar to a body weight of 18-22 g, half male and half female, fasted for 24 hours before the experiment, and intraperitoneally injected with streptozotocin (STZ) 50 mg / kg (1% citrate buffer). liquid solution preparation). One week later, blood glucose (BS) was measured by blood collection from the tail vein of the mice. If the BS was higher than 16mmol / L, the modeling was successful. After modeling, the mice were divided into 3 groups, 10 in each group. Diabetes model group (DM group): subcutaneous injection of the same amount of PBS buffer; control group (Bai Sugar): 20 μg / kg subcutaneous injection dissolved in 0.1mol / L PBS buffer), twice a day for 15 consecutive days; sample group (silkworm chrysalis polypeptide 1): the same as the control group; another 10 normal mice were taken as the blank group. On the 15th day after administration, blood was...

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Abstract

The invention relates to the field of medicine and in particular relates to silkworm polypeptide 1 with capability of preventing or treating diabetes mellitus. The sequence of the silkworm polypeptide 1 is RLGRRLSEDPATPADQ. A preparation method of the silkworm polypeptide 1 comprises the following steps: (1) decocting; (2) enzymatic hydrolysis; (3) protein separation; (4) semi-preparative RP-HPLC separation and purification; and (5) freeze-drying, wherein papain and alkaline protease are adopted to carry out enzymatic hydrolysis; macroporous resin and sephadex are adopted to carry out protein separation. The invention also relates to application of the silkworm polypeptide 1 in treatment of the diabetes mellitus and complications of the diabetes mellitus. The complications of the diabetes mellitus include diabetic nephropathy, diabetic hypertension, diabetic eye disease and diabetic neurologic disorder. Since polypeptide is extracted from silkworm, the silkworm polypeptide 1 disclosed by the invention is good in safety and is capable of effectively preventing or treating the diabetes mellitus.

Description

Technical field: [0001] The invention relates to the field of medicine, in particular to a polypeptide extract for treating diabetes. Background technique [0002] Diabetes mellitus is a group of metabolic diseases characterized by hyperglycemia. It is a syndrome of glucose, protein and lipid metabolism disorders caused by insufficient insulin or defects in insulin cell metabolism. In recent years, the number of diabetic patients has gradually increased, and is divided into type I diabetes and type II diabetes. Among them, the number of patients with type II diabetes accounts for 85-90% of the total number of diabetes, which seriously threatens the health and life of patients. If diabetes is not well controlled, it may cause many complications, such as diabetic nephropathy, retinopathy, high blood pressure, etc. [0003] At present, most patients treat diabetes by injecting insulin or oral hypoglycemic drugs to help control blood sugar in the body, or regulating insulin se...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08A61K38/10A61P3/10A61P13/12A61P9/12A61P27/02A61P25/00
Inventor 罗瑞雪
Owner SUZHOU PULUODA BIOLOGICAL SCI & TECH
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