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A kind of cell screening model of diuretic activity drug and its establishment method and application

A method for establishing a method and a technology for active drugs, applied in the field of pharmacy, can solve the problems of long time, high cost, difficult operation, etc., and achieve the effect of reducing the concentration of chloride ions and inhibiting the transcellular transport of chloride ions

Active Publication Date: 2022-03-15
贵州中医药大学
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above-mentioned animal water diuresis model screening method is costly, takes a long time, and is not easy to operate

Method used

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  • A kind of cell screening model of diuretic activity drug and its establishment method and application
  • A kind of cell screening model of diuretic activity drug and its establishment method and application
  • A kind of cell screening model of diuretic activity drug and its establishment method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Method for Establishing Cellular Screening Model of Diuretic Active Drugs

[0047] 1. Culture of MDCK cells

[0048] Purchase Madin-Daby canine kidney cells (MDCK) cell line, use DMEM high-glucose medium containing 5% fetal bovine serum, 0.5% penicillin-streptomycin solution, 94.5%, 37°C, CO 2 MDCK cells were cultured in the incubator until the logarithmic growth phase.

[0049] 2. Drug preparation

[0050] Take NaCl, DMEM high-glucose medium into 0.012, 0.015, 0.018, 0.021, 0.024, 0.027, 0.030, 0.033, 0.036g·mL -1 NaCl is set aside. Relebene, 22-hydroxyhobopentane, β-sitosterol, cycloeucalenone, zeugatene, dryocrassol, Hopan-28,22-olide, oleanolic acid, 22,28-epoxyhopane, cyclohopenol, cycloeucalenol, suberinone , (23E)-cycloart-23-ene-3β,25-diol, (23E)-cycloart-23,25-ene-3β-ol, cyclolaudenol, 11-carbonyl-β-acetylboswellic acid, 2',3' -dihydroxypropylpentadecanoate, β-carotin, and hydrochlorothiazide were dissolved in DMSO, and then diluted to 100umol L in DMEM hi...

Embodiment 2

[0059] Method for Establishing Cellular Screening Model of Diuretic Active Drugs

[0060] 1. Culture of MDCK cells

[0061] Purchase Madin-Daby canine kidney cells (MDCK) cell line, use 15% fetal bovine serum, 1.5% penicillin-streptomycin solution, 83.5% DMEM high-glucose medium, 37°C, CO 2 MDCK cells were cultured in the incubator until the logarithmic growth phase.

[0062] 2. Drug preparation

[0063]Take NaCl, DMEM high-glucose medium into 0.012, 0.015, 0.018, 0.021, 0.024, 0.027, 0.030, 0.033, 0.036g·mL -1 NaCl is set aside. Relebene, 22-hydroxyhobopentane, β-sitosterol, cycloeucalenone, zeugatene, dryocrassol, Hopan-28,22-olide, oleanolic acid, 22,28-epoxyhopane, cyclohopenol, cycloeucalenol, suberinone , (23E)-cycloart-23-ene-3β,25-diol, (23E)-cycloart-23,25-ene-3β-ol, cyclolaudenol, 11-carbonyl-β-acetylboswellic acid, 2',3' -dihydroxypropylpentadecanoate, β-carotin, and hydrochlorothiazide were dissolved in DMSO, and then diluted to 100umol L in DMEM high-glucose ...

Embodiment 3

[0072] Method for Establishing Cellular Screening Model of Diuretic Active Drugs

[0073] 1. Culture of MDCK cells

[0074] Purchase Madin-Daby canine kidney cells (MDCK) cell line, use 12.3% fetal bovine serum, 0.7% penicillin-streptomycin solution, 87% DMEM high glucose medium, 37°C, CO 2 MDCK cells were cultured in the incubator until the logarithmic growth phase.

[0075] 2. Drug preparation

[0076] Take NaCl, DMEM high-glucose medium into 0.012, 0.015, 0.018, 0.021, 0.024, 0.027, 0.030, 0.033, 0.036g·mL -1 NaCl is set aside. Relebene, 22-hydroxyhobopentane, β-sitosterol, cycloeucalenone, zeugatene, dryocrassol, Hopan-28,22-olide, oleanolic acid, 22,28-epoxyhopane, cyclohopenol, cycloeucalenol, suberinone , (23E)-cycloart-23-ene-3β,25-diol, (23E)-cycloart-23,25-ene-3β-ol, cyclolaudenol, 11-carbonyl-β-acetylboswellic acid, 2',3' -dihydroxypropylpentadecanoate, β-carotin, and hydrochlorothiazide were dissolved in DMSO, and then diluted to 100umol L in DMEM high-glucose...

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Abstract

The present invention relates to a cell screening model of diuretic drug and its establishment method and application. The method comprises the following steps: 1) culturing of MDCK cells; 2) preparation of drugs; 3) screening of NaCl transport concentration; 4) grouping, giving Drug and NaCl transport experiment; 5) Statistical analysis. This method adopts a model of renal tubular cells simulating sodium chloride transport to screen diuretic active drugs, and the diuretic active screening experiments on monomers extracted from Shiwei prove to be effective, and it is a relatively fast cell model diuretic drug screening model.

Description

technical field [0001] The invention relates to the field of pharmacy, in particular to a cell screening model of diuretic active drugs and its establishment method and application. Background technique [0002] According to the existing literature, drug screening refers to the process of evaluating the biological activity, pharmacological effect and medicinal value of substances (sampling) that may be used as drugs by using appropriate methods. Drug screening is screening at the biochemical and cellular levels. [0003] The screening model is the pharmacological experimental model used in drug screening experiments. Since drug screening requires standardized and quantitative characteristics of the experimental protocol, animal experiments that are common in traditional pharmacological experiments are rarely used in drug screening. According to the experimental model Drug screening can be divided into screening at the biochemical level and screening at the cellular level. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071C12Q1/02
CPCC12N5/0686G01N33/5008
Inventor 杨武德隋怡龙毅罗国勇於祥
Owner 贵州中医药大学
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