Method for restraining TNF-alpha induced human glomerular mesangial cell excessive proliferation and inflammatory pathway

A technology of mesangial cells and glomeruli, applied in the field of pharmacy, can solve the problems of hypocalcemia, patients prone to allergic reactions, infections, etc., and achieve the effect of obvious technical advantages.

Inactive Publication Date: 2019-10-25
YANTAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Plasma exchange has a good effect on patients with LN, but during treatment, patients are prone to severe complications such as allergic reactions, infections, hypokalemia, and hypocalcemia, and this method is not suitable for patients with hemolytic uremia LN patients with vasculitis or vasculitis

Method used

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  • Method for restraining TNF-alpha induced human glomerular mesangial cell excessive proliferation and inflammatory pathway
  • Method for restraining TNF-alpha induced human glomerular mesangial cell excessive proliferation and inflammatory pathway
  • Method for restraining TNF-alpha induced human glomerular mesangial cell excessive proliferation and inflammatory pathway

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Method for Inhibiting the Hyperproliferation of Human Glomerular Mesangial Cells Induced by TNF-α

[0046] Human glomerular mesangial cells in the logarithmic growth phase were digested with trypsin and made into a single-cell suspension. At the same time, the 1 μg / mL TNF-α stock solution was diluted to a final concentration of 10 ng / mL with sterile PBS, and then Add TNF-α at a concentration of 10 ng / mL so that about 50 μL of TNF-α per 8000 cells, and then stimulate the human glomerular mesangial cells for 24 hours;

[0047] Temperature is 37°C, humidity is 95%, CO 2 The concentration is 5%, under sterile conditions, the human glomerular mesangial cells induced by TNF-α are respectively added with quercetin at a concentration of 20 μM, 40 μM, and 80 μM for 24 hours, and a blank control group (control), In the TNF-α group, the inhibitory effect was verified by control under the same conditions;

[0048] Then, measure the OD value under the wavelength of 570nm...

Embodiment 2

[0052] Example 2 The method for inhibiting the excessive proliferation of human glomerular mesangial cells induced by TNF-α

[0053] The operation process is the same as in Example 1, and a blank control group (control) and a TNF-α group are also respectively set up. The difference is that under the same conditions, a concentration of 80 μM quercetin is added to the human glomerular mesangial cells induced by TNF-α. Table 1 shows the results after 6h, 12h, 24h, and 48h of prime treatment respectively.

[0054] Table 1. Results of quercetin 80μM intervention for 6h, 12h, 24h, 48h

[0055]

[0056] Such as figure 2 The results showed that quercetin at a concentration of 80 μM inhibited the proliferation of human glomerular mesangial cells within a time period of 6h-48h to a level close to that of the blank group, and the inhibitory effect gradually increased with the prolongation of the action time.

Embodiment 3

[0057] Example 3 Verification of the inhibitory effectiveness of Qu on the excessive proliferation of human glomerular mesangial cells

[0058] The experiment was divided into five groups: blank control group, TNF-α group, TNF-α+Qu 20μM group, TNF-α+Qu 40μM group, TNF-α+Qu 80μM group. Three parallel wells were set up for each experiment in each group.

[0059] The test steps are as follows:

[0060] 1) Cell inoculation: Human glomerular mesangial cells in the logarithmic growth phase were taken, digested with trypsin, made into a single cell suspension, and counted under a microscope. In the process of cell counting, follow the counting principle of "counting up but not down, counting left but not right, and counting one group when encountering one". Then use the counting formula to calculate, the cell counting formula is as follows:

[0061] The number of cells in the cell suspension / mL=the total number of cells in the four grids / 4×10 4 ×Dilution factor

[0062] Accordin...

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Abstract

The invention discloses a method for restraining TNF-alpha induced human glomerular mesangial cell excessive proliferation and an inflammatory pathway. The method includes the following steps that intervention of quercetin with the concentration being 20 microns-80 microns is conducted on TNF-alpha induced human glomerular mesangial cells with the final concentration being 5-20 ng/mL for 6-48 hours, so that TNF-alpha induced human glomerular mesangial cell proliferation is effectively restrained. According to the method for restraining TNF-alpha induced human glomerular mesangial cell proliferation, the NF-kB signal pathway is interdicted through quercetin Qu to reduce PTX3 expression so as to restrain LN, the method has a remarkable advantages in the pathological change aspects of protecting LN and improving mesangial cell excessive proliferation, and compared with usage of glucocorticoid and other medicine, quercetin has the advantage of being free of side effects.

Description

technical field [0001] The invention relates to a method for inhibiting excessive proliferation and inflammatory pathways of LN mesangial cells at the level of glomerular mesangial cells, and belongs to the field of pharmacy. Background technique [0002] Lupus Nephritis (LN) is nephritis caused by the autoimmune disease Systemic Lupus Erythematosus (SLE), with a high fatality rate. About 70% of the patients had different degrees of clinical manifestations of renal damage, with varying degrees of proteinuria and microscopic hematuria being the most common, often accompanied by urinary casts and renal dysfunction. Among patients with systemic lupus erythematosus, as many as 60% will develop lupus nephritis, and about 10% to 15% of patients with lupus nephritis will eventually develop into end-stage renal failure, which seriously reduces the survival rate of patients. Therefore, in the process of treating systemic lupus erythematosus, how to control the occurrence and develop...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/352A61P13/12G01N33/68
CPCA61K31/352A61P13/12G01N33/6893
Inventor 范华英刘悦冀凯刘珂李新王学凯杜世豪王宇王梦影
Owner YANTAI UNIV
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