Microfluidic chip-based detection of influence of pentoxifylline on erythrocyte deformability and biochemical indexes in patients with coronary heart disease

A microfluidic chip, pentoxifylline technology, applied in biological testing, material inspection products, testing moving fluid/granular solids, etc., to achieve the effect of easy operation, low sample consumption, and good portability

Inactive Publication Date: 2018-08-24
SOUTHERN MEDICAL UNIVERSITY
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  • Description
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Problems solved by technology

Although several studies have proposed the effect of PTX on the deformability of RBC in vitro, the individual differences and causes of PTX on the deformability of red blood cells in patients with coronary heart disease in vitro have been systematically explored, and by combining the mechanical properties of red blood cells with biochemical parameters , including Na + -K + -ATPase, superoxide dismutase activity (SOD), malondialdehyde content (MDA), cholesterol content, sialic acid content (SA), hemoglobin content (Hb) and membrane protein skeleton components are analyzed in combination to find the main influence Studies on biochemical indicators have not been reported

Method used

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  • Microfluidic chip-based detection of influence of pentoxifylline on erythrocyte deformability and biochemical indexes in patients with coronary heart disease
  • Microfluidic chip-based detection of influence of pentoxifylline on erythrocyte deformability and biochemical indexes in patients with coronary heart disease
  • Microfluidic chip-based detection of influence of pentoxifylline on erythrocyte deformability and biochemical indexes in patients with coronary heart disease

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Experimental program
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Effect test

Embodiment 1

[0057] Embodiment 1 Fabrication of microfluidic chip

[0058] Fabrication of microfluidic chips: using SU-8 photoresist and standard wet etching process to prepare PDMS microchannel templates on silicon wafers, and adopt casting method to prepare PDMS microchannel chips. The preparation method includes the following steps:

[0059] First, silanize the silicon wafer template with 25mL trimethylchlorosilane solution in a vacuum pump for 1 hour to prevent PDMS from sticking, and it is easier to tear off the PDMS glue, and clean it ultrasonically with acetone, ethanol and ultrapure water, and then blow it with nitrogen. Dry and dry. Then take PDMS and curing agent with a weight ratio of 10:1 and mix them well, put them in a vacuum desiccator to degas until the bubbles completely disappear, and then pour them slowly on the silicon wafer to avoid bubbles. Then it was cured on a heating plate at 95°C for 3 hours, and then the PDMS was peeled off from the silicon wafer, so that the m...

Embodiment 2

[0061] experimental method:

[0062] 1. Blood Sample Preparation

[0063] Blood samples were collected from the veins of patients with coronary heart disease, and the fresh blood was centrifuged at 300g for 3 minutes to remove the supernatant. Then it was equally divided into four parts and added to RPMI 1640 medium containing 20% ​​fetal bovine serum (FBS), so that the final volume of RBC solution was 1%. Then, PTX was added to the RBC solution to make the final concentration of 0, 2, 20, and 200 μg / mL, and incubated at 37°C for 2 hours. After incubation, RBC samples were washed 3 times with RPMI solution containing 20% ​​FBS to remove excess PTX, and then stored at 4°C for future use.

[0064] 2. Measurement of red blood cell deformability

[0065] Take 10 μL of red blood cells incubated with drug (PTX), mix them with DilC18(3) cell orange fluorescent dye with a final concentration of 10 μM, and incubate at 37°C for 15 minutes to stain the red blood cells. Red blood cell...

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Abstract

The invention discloses microfluidic chip-based detection of influence of pentoxifylline on erythrocyte deformability and biochemical indexes in patients with coronary heart disease. The individualized detection of the influence of the pentoxifylline on the erythrocyte deformability is realized based on a microfluidic chip, detection and screening of innovative drugs for improving the erythrocytedeformability can be realized, and three main effective biochemical indicators of Na<+>-K<+>-ATPase, superoxide dismutase activity and cholesterol content are found to be important in the change of the erythrocyte deformability in the patients with coronary heart disease.

Description

technical field [0001] The invention relates to a method for detecting the influence of pentoxifylline on red blood cell deformability and biochemical indexes of patients with coronary heart disease based on a microfluidic chip. Background technique [0002] Human red blood cells (RBCs) are elastic biconcave disc cells with a diameter of about 8 μm, which are mainly responsible for the transport of oxygen and carbon dioxide. Erythrocyte deformability refers to the ability of erythrocytes to maintain microcirculation by changing their shape to pass through microcirculatory capillaries as small as 3 μm in diameter. The reduced deformability of red blood cells can lead to microcirculatory disturbance, increased blood viscosity, and red blood cells with reduced deformability can block capillary blood flow and lead to ischemic tissue necrosis. Reduced red blood cell deformability is also a marker of certain underlying diseases. Considering the importance of RBC deformability, s...

Claims

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Application Information

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IPC IPC(8): G01N33/48G01N21/85
CPCG01N33/48G01N21/85
Inventor 刘利红刘贞
Owner SOUTHERN MEDICAL UNIVERSITY
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