Diuretic active drug cell screening model, and establishing method and application thereof

A technology for establishing methods and active drugs, applied in the field of pharmacy, can solve the problems of high cost, long time, and difficult operation.

Active Publication Date: 2020-09-22
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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  • Diuretic active drug cell screening model, and establishing method and application thereof
  • Diuretic active drug cell screening model, and establishing method and application thereof
  • Diuretic active drug cell screening model, and establishing method and application thereof

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 calf 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 invention relates to a diuretic active drug cell screening model, and an establishing method and application thereof. The method comprises the steps: (1), culture of MDCK cells; (2), preparation of drugs; (3), screening of NaCl transport concentration; (4), grouping, drug administration and NaCl transport experiment; and (5), statistical analysis. According to the method in the invention, diuretic active drugs are screened through a model for simulating sodium chloride transport by renal tubular cells is adopted; furthermore, the fact that it is effective can be proved in a pyrrosia linguaextraction monomer diuretic active screening experiment; and the model is a relatively rapid 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 Applications(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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