Application of linarin to preparing kidney tubular epithedial cell ischemia reperfusion injury protecting drugs/drug combinations

An ischemia-reperfusion and epithelial cell technology, which can be used in drug combinations, pharmaceutical formulations, and medical preparations containing active ingredients, etc. It can solve the problem of unclear role of ETS2 protein, and achieve the effect of broadening the scope of application.

Active Publication Date: 2019-06-21
THE AFFILIATED HOSPITAL OF QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At this stage, although the research on Menghuaside is extensive, its effect on ETS2 protein and whether it has a protective effect on renal ischemia-reperfusion injury are still unclear

Method used

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  • Application of linarin to preparing kidney tubular epithedial cell ischemia reperfusion injury protecting drugs/drug combinations
  • Application of linarin to preparing kidney tubular epithedial cell ischemia reperfusion injury protecting drugs/drug combinations
  • Application of linarin to preparing kidney tubular epithedial cell ischemia reperfusion injury protecting drugs/drug combinations

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 HK2 cell ischemia-reperfusion model

[0024] Plant HK2 cells into a 35mm cell culture dish and culture in 2mL complete medium (89% DMEM + 10 fetal bovine serum + 1% penicillin streptomycin mixture), 5% carbon dioxide, and a humid environment to 70% confluence At the same time, the cells were divided into the model group and the control group. The cells in the model group were removed from the complete medium, washed twice with PBS, and then added 2 mL of sugar-free and serum-free DMEM medium (containing 1% penicillin-streptomycin mixed solution) into 1 %Oxygen, 5% carbon dioxide, and 94% nitrogen were cultured in a humid environment for 3h, 6h, 12h, 24h, 36h, and 48h respectively, while the cells in the control group were sucked out of the original medium, washed twice with PBS, and then added 2mL of complete medium , cultured in 5% carbon dioxide, normal oxygen, and humid environment for the same time. After the hypoxic culture, the model group was replaced w...

Embodiment 2

[0025] ETS2mRNA level detection in the model of embodiment 2

[0026] 2.1 mRNA extraction

[0027] Add 200 μL of chloroform to the collected trizol and cell lysate solution per milliliter of trizol, mix manually for 1 minute, place on ice for 3 minutes, then centrifuge at 12,000 rpm at 4°C, collect the supernatant, add an equal volume of isopropanol, and place at 4°C for 10 minutes After centrifugation, discard the supernatant and add 1mL of 75% ethanol, vortex, centrifuge again, absorb the supernatant, add appropriate amount of DEPC water to dissolve the liquid after drying, and finally use a spectrophotometer to detect the RNA concentration. The OD260 / OD280 ratios of the extracted RNAs were all between 1.8 and 2.0.

[0028] 2.2 Reverse transcription, real-time fluorescent quantitative PCR

[0029] TAKARA reverse transcription kit (RR037a) was used to perform reverse transcription according to its instructions, and the obtained cDNA was used as a template for real-time fluo...

Embodiment 3

[0031] Example 3 The Toxic Effect of Menghuaside on HK2 Cells

[0032] The same amount of HK2 cells was planted into each well of a 96-well plate, and when the cells reached 70% confluence, they were replaced with serum-free DMEMf12 medium, and 5 μM, 10 μM, 30 μM, 50 μM, 70 μM, and 100 μM mongoside were added to different wells respectively. Cultivate for 48h. After the cultivation, the liquid in the wells was aspirated, and 10 μL of CCK8 working solution and 100 μL of DMEM f12 medium were added to each well, and the absorbance at 560 nm was detected in a microplate reader after incubation at 37 °C for 4 h. The toxic effect of mongoside was judged by calculating the cell viability.

[0033] The result shows, as figure 2As shown, at the concentration > 50 μM, the activity of HK2 cells began to decline, but the maximum non-toxic concentration of succulentin to heart cells in previous studies was 30 μM (cultured for 48 h), considering the application of succulentin in human bo...

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Abstract

The invention discloses application of linarin to preparing kidney tubular epithedial cell ischemia reperfusion injury protecting drugs / drug combinations and belongs to the field of biological medicines. The application proves the possibility of protecting effects of the linarin on kidney ischemia reperfusion, and through data, discovers that reducing expression of ETS2 proteins can inhibit proinflammatory factors IL-12 and accordingly inhibit inflammation to achieve the protecting effects on kidney tubular epithedial cell ischemia reperfusion injury; further only through the molecule dockingtechnology, possibility of interaction between the linarin and the ETS2 proteins is analyzed, and a theoretical foundation for enlarging the possible application range of the linarin is provided. Theapplication of the linarin to preparing the kidney tubular epithedial cell ischemia reperfusion injury protecting drugs / drug combinations can be applied to theoretical research and clinical treatmentof kidney ischemia reperfusion injury and provide the theoretical foundation for enlarging the possible application range of the linarin.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to the application of fenugreek in the preparation of a drug / medicine composition for protecting renal tubular epithelial cells from ischemia-reperfusion injury. Background technique [0002] Acute kidney injury (Acute Kidney Injury, AKI) is a common clinical critical disease. Studies have shown that the mortality rate of AKI is as high as 50% to 79%, and the proportion of survivors who develop into chronic renal failure is as high as 35% to 71%. Injury is one of the most common causes of AKI. A large number of animal model experiments and human histopathological studies of renal ischemia-reperfusion injury have shown that inflammation is the most important pathophysiological change of ischemia-reperfusion acute kidney injury. [0003] Menghuaside (CAS: 480-36-4) is the extract of wild chrysanthemum flower, its chemical formula is C 28 h 32 o 14 , with a molecular weight o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7048A61P13/12
Inventor 徐岩杨成宇蒋伟
Owner THE AFFILIATED HOSPITAL OF QINGDAO UNIV
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