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Autophagy revulsive for diabetic vascular complication treatment and application in medicine

A technology of diabetes blood vessels and inducers, applied in the field of biomedicine, can solve the problems of application of diabetes and vascular complications, etc.

Inactive Publication Date: 2017-02-22
JIANGSU PROVINCE INST OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this compound has not been used in the treatment of diabetes and vascular complications

Method used

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  • Autophagy revulsive for diabetic vascular complication treatment and application in medicine
  • Autophagy revulsive for diabetic vascular complication treatment and application in medicine
  • Autophagy revulsive for diabetic vascular complication treatment and application in medicine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 MTT detection of HK-2 cell viability

[0026] 1. Instruments and reagents:

[0027] Human proximal tubular epithelial cell line (HK-2); DMEM (low glucose) medium, fetal bovine serum, trypsin, PBS buffer, CO 2 Cell incubator, digital display constant temperature stirring circulating water bath, cell culture ultra-clean bench, low temperature ultracentrifuge, inverted microscope, micropipette, automatic microplate reader.

[0028] 2. Experimental method

[0029] (1) In vitro culture of HK-2 cells

[0030] Cultured with low-sugar DMEM+10% FBS medium at 37°C, 5% CO 2 Cells were passaged when grown under conditioned conditions to 80%-90% confluency. Pour off the medium in the culture vessel, wash the cells once with PBS, discard the PBS, add an appropriate amount of 0.1% trypsin solution according to the culture area to digest, place at 37°C for 3-5min, and observe under an inverted microscope until the cells reach a maximum size. After part of the cells becam...

Embodiment 2

[0036] Example 2 Detection of autophagy in HK-2 cells by transmission electron microscopy and immunofluorescence

[0037] 1. Instruments and reagents:

[0038] Human proximal renal tubular epithelial cell line (HK-2); DMEM (low glucose) medium, fetal bovine serum, trypsin, PBS buffer, glutaraldehyde, osmic acid, acetone, embedding solution, 3% uranyl acetate- Lead citrate, acridine orange stain, CO 2 Cell incubator, digital display constant temperature stirring circulating water bath, cell culture ultra-clean bench, low-temperature ultracentrifuge, transmission electron microscope, ultra-thin slicer, inverted fluorescence microscope.

[0039] 2. Experimental method

[0040] (1) Detection of autophagy in HK-2 cells by transmission electron microscopy

[0041] HK-2 cells were cultured according to the method described in Example 1, and the HK-2 cells in the logarithmic growth phase were divided into 2 × 10 6 Each cell / bottle was evenly inoculated into a culture dish with a d...

Embodiment 3

[0046] Example 3 Western blot detection of autophagy-related protein expression

[0047] 1. Instruments and reagents

[0048] Human proximal renal tubular epithelial cell line (HK-2); DDMEM (low glucose) medium, fetal bovine serum, trypsin, PBS buffer, bovine serum albumin (BSA), tissue protein lysate, BCA protein concentration determination reagent Box, ECL, SDS, Ponceau, PVDF membrane; Antibodies: rabbit anti-human LC3 polyclonal antibody, rabbit anti-human Atg-5 monoclonal antibody, rabbit anti-human Atg-12 monoclonal antibody, rabbit anti-human Beclin-1 monoclonal antibody, rabbit anti-human β-actin polyclonal antibody; horseradish peroxidase-labeled goat anti-rabbit secondary antibody, neutral gum, hematoxylin, DAB, ethanol, 3% hydrogen peroxide, petri dish , CO 2 Cell incubator, digital display constant temperature stirring and circulating water bath, cell culture ultra-clean workbench, low temperature ultracentrifuge, inverted microscope, micro pipette, SDS-PAGE gel e...

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PUM

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Abstract

The invention discloses a new application for alisol B autophagy revulsive for treating diabetic and vascular complication, and the application in medicine. The invention through the experiment methods of electron microscope transmission, immunofluorescence testing, Westernblot, and more, fully proves the functions of alisol B in controlling the autophagic disorders of human kidney tubular epithelial cells, vascular endothelial cells, retinal cells, and cardiac muscle cells caused by advanced glycated end products (AGEs). The invention discovers the marker and action mechanism of alisol B inducer cell autophagy. Therefore, the proposal applies alisol B in the treatments of autophagy action mechanism related diseases, and particularly has broad application prospects in diabetics induced by autophagic disorders and in treatments of vascular complications as well as medicine developments.

Description

technical field [0001] The invention relates to alisol B as a natural active compound as an autophagy inducer and its application in treating diabetic vascular complications. The invention belongs to the field of biomedicine. Background technique [0002] Diabetes has become one of the common diseases in the modern population, seriously affecting human health and living health. Diabetes is not only reflected in the level of blood sugar in the body, but more importantly, vascular complications caused by diabetes have become an important reason affecting health. According to the latest epidemiological survey, there are currently about 114 million people with diabetes diagnosed in China, and about 140 million people with pre-diabetes. In the diabetic population, there are more or less one or more complications, such as nephropathy and retinopathy. According to research, more than 90% of patients with diabetes have complications for more than 10 years. The prevention and tre...

Claims

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Application Information

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IPC IPC(8): A61K31/58A61P3/10A61P13/12A61P9/00A61P9/10A61P27/02
CPCA61K31/58
Inventor 封亮贾晓斌汪春飞汪刚刘莹谈唯
Owner JIANGSU PROVINCE INST OF TRADITIONAL CHINESE MEDICINE
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