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Method for screening diabetic nephropathy drug in vitro

An in vitro screening technology for diabetic nephropathy, which is applied in the field of screening drugs for diabetic nephropathy, can solve the problems of many uncontrollable factors, long time for DN animal model modeling, and unstable models, so as to reduce economic and time costs and improve the efficiency of new drug screening Effect

Active Publication Date: 2017-12-15
BEIJING HANDIAN PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The problems of using this model include: 1) high mortality rate of animals; 2) relatively mild renal lesions, less likely to form glomerulosclerosis, tubulointerstitial fibrosis and other renal pathological changes similar to human DN patients in the middle and late stages; 3) STZ has nephrotoxicity, which will interfere with the experimental results to a certain extent; 4) The severity of proteinuria is related to the dose of STZ
[0009] In summary, DN animal models generally have problems such as long modeling time, cumbersome operation, high cost, and unstable models.
For the drug screening and drug efficacy evaluation of DN new drug development, it takes at least half a year, costs hundreds of thousands to more than one million yuan, and there are many uncontrollable factors

Method used

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  • Method for screening diabetic nephropathy drug in vitro
  • Method for screening diabetic nephropathy drug in vitro
  • Method for screening diabetic nephropathy drug in vitro

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1 Screening experiment on the preparation process of astragalus gorgon rhubarb traditional Chinese medicine composition

[0050] The purpose of the experiment: to screen the optimal preparation process of the traditional Chinese medicine composition through the method of serum pharmacology, combined with the in vitro drug efficacy screening method of the present invention, so as to lay the foundation for subsequent research and development.

[0051] 1. Experimental materials

[0052] (1) Experimental animals

[0053] 30 SPF grade SD rats, 180-220g, were purchased from Beijing Huafukang Biotechnology Co., Ltd., license number: SCXK (Beijing) 2014-0004, and were bred in Beijing Yizhuang International Biomedical Technology Co., Ltd., license number : SYXK (Beijing) 2015-0010.

[0054] (2) Experimental cells

[0055] Human renal tubular epithelial cells HKC were purchased from the Cell Resource Center of the Institute of Basic Medical Sciences, Chinese Academy of...

Embodiment 2

[0100] Embodiment 2 rhubarb preparation technology screening experiment

[0101] The results of Example 1 show that the drug effect of process 4 is better than that of process 3. The difference between process 4 and process 3 is only that the extraction solvent of rhubarb is different. In process 3, rhubarb is alcohol extraction, and in process 4, rhubarb is water extraction. In this experiment The glomerular mesangial cells and renal tubular epithelial cells most closely related to the onset of diabetic nephropathy were selected to test whether the water-extracted sample of rhubarb was more effective than the alcohol-extracted sample of rhubarb.

[0102] 1. Experimental materials

[0103] (1) Experimental animals

[0104] Animals 18 SPF grade SD rats, 180-220g, were purchased from Beijing Huafukang Biotechnology Co., Ltd., license number: SCXK (Beijing) 2014-0004, and were raised in Beijing Yizhuang International Biomedical Technology Co., Ltd., license No.: SYXK (Beijing) ...

Embodiment 3

[0149] Example 3 Astragalus gorgon rhubarb traditional Chinese medicine composition process 3 and process 4 animal experiments

[0150] The results of Examples 1 and 2 show that Process 4 is superior to Process 3, and even better than Processes 1 and 2 in terms of inhibiting proliferation of renal intrinsic cells and increasing glucose intake under high glucose conditions. In order to further confirm the reliability of the proposed screening method, the in vivo animal experiment was conducted again through the type 1 SD rat DN model to test the conclusion of the in vitro cell screening test.

[0151] 1. Experimental materials

[0152] (1) Experimental animals

[0153] 57 SPF grade SD rats, 180-220g, were purchased from Beijing Huafukang Biotechnology Co., Ltd., license number: SCXK (Beijing) 2014-0004, and were bred in Beijing Yizhuang International Biomedical Technology Co., Ltd., license number : SYXK (Beijing) 2015-0010.

[0154] (2) Experimental drugs

[0155] Prescrip...

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Abstract

The invention provides a method for screening a diabetic nephropathy drug in vitro by determining the change of glucose concentration in culture solution of a unit cell of glomerular mesangial cells, renal tubular epithelial cells and endothelial cells after the drug is administrated and the change of the cellular proliferation inhibition rate. The method is not only suitable for screening of small molecule compounds, but also especially suitable for screening of prescription compatibility of traditional Chinese medicine compositions and screening of preparation processes. When a model and the method are applied to screening of the drug, the cost is low, the time is short, and the operation is simple, convenient and easy and is strong in reliability, therefore, the method has the important significance for early screening of drug candidates for diabetic nephropathy.

Description

technical field [0001] The invention relates to a method for screening drugs in vitro, which is mainly used for screening drugs for diabetic nephropathy. More specifically, the present invention relates to a method for screening diabetic nephropathy drugs in vitro using high-throughput means. Background technique [0002] Diabetic nephropathy (diabetic nephropathy, DN) is currently one of the most serious and common complications of diabetes. The blood vessels of patients are continuously in a state of high glucose, resulting in various microvascular diseases. The early manifestations of DN are renal extracellular matrix hyperplasia, glomerular hyperfiltration and renal tubular hyperperfusion. As the course of the disease progresses, it causes progressive loss of glomerulus and tubular cells, eventually leading to glomerulosclerosis, tubular atrophy, and renal interstitial fibrosis. Typical pathological manifestations are: increased proteinuria; a transient increase in glo...

Claims

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Application Information

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IPC IPC(8): C12Q1/02C12Q1/26C12Q1/28C12Q1/54
CPCC12Q1/26C12Q1/28C12Q1/54G01N33/5044G01N2800/347
Inventor 林德良蒿长英钟云黄卉
Owner BEIJING HANDIAN PHARMA CO LTD
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