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Establishment method and application of renal aging model

A technology for establishing methods and senescent cell models, applied in biochemical equipment and methods, screening of compounds, urinary tract/kidney cells, etc., can solve problems such as there is no D-galactose kidney aging model

Inactive Publication Date: 2018-05-04
NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
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  • Abstract
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  • Claims
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Problems solved by technology

However, there is no report on the use of D-galactose to establish a model of renal aging

Method used

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  • Establishment method and application of renal aging model
  • Establishment method and application of renal aging model
  • Establishment method and application of renal aging model

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Embodiment 1

[0023] 1. Materials and Reagents

[0024] 1.1 Experimental cells and reagents

[0025] Rat renal tubular epithelial cell lines were provided by Shanghai Cell Bank; PBS buffer, DMEM medium (full name dulbecco's modified eagle medium, which is a medium containing various amino acids and glucose), and streptomycin-penicillin solution were purchased from Hyclone Company; 0.25% trypsin, purchased from Gibco Company; fetal bovine serum (FBS), purchased from Zhejiang Tianhang Biotechnology Co., Ltd.; thiazolyl blue (MTT), purchased from BIOSHARP Sigma Company; D-galactose (cell grade , Lot#: WXBC1780V), purchased from Sigma Company; cell senescence β-galactosidase staining kit (Lot: 0314161600503), purchased from Shanghai Beyond Biotechnology Co., Ltd.; fluorescein di-β-D-pyransemi Lactose (FDG, Lot#:025029) was purchased from AATBioquest; Fluorescein (AR, Lot#:C10072457) was purchased from McLean; SOD(Lot:20170310), MDA(Lot:20170107), GSH-PX(Lot : 20170329) kits were purchased fro...

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Abstract

The invention discloses an establishment method of a renal aging model. The establishment method comprises the following steps: (1), culturing rat renal tubular epithelial cells in an FBS containing DMEM (dulbecco's modified eagle medium) culture medium at 37 DEG C in presence of CO2 with the concentration of 5%, and after growing to 85-90%, performing subculture according to a ratio of 1:4, wherein cells of each generation are kept and cryopreserved; taking 9th-generation logarithmic phase cells, adjusting the cell concentration to (1-5)*10<4> mL<-1>, and then adding into a 96-mesh plate; (2), adding 10-20 mg.mL<-1> of D-galactose into the 96-mesh plate, and culturing for 24-48 hours to obtain the renal aging model. Experimental results show that the SOD and GSH-PX activities of cells inan induction group have a time-dependent change along with the MDA content under the action of the D-galactose, the SOD and GSH-PX activities gradually decrease and the MDA content gradually increases; the renal aging model established according to the establishment method provided by the invention can be used for screening medicines for preventing and treating renal aging or failure and is of great significance.

Description

technical field [0001] The invention relates to a method for establishing a model, in particular to a method for establishing a renal aging model, which can be used to screen drugs for resisting renal aging or failure. Background technique [0002] With the advent of an aging society, there are more and more studies on how to fight aging. There are also many methods for constructing cell aging models in vitro, such as H2O2 oxidative damage aging, X-ray induced aging, and continuous passage in vitro for natural aging. D-galactose is often used as a food additive of sweetener, and it is also contained in many dairy products in daily life. Studies in the 1960s found that galactose had deleterious effects on energy and phospholipid metabolism in the chicken brain. Chinese scholar Xu Fuben first proposed that administration of D-galactose to rodents can induce physiological changes similar to accelerated aging process, and produce physiological and biochemical changes such as d...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12Q1/02
CPCC12N5/0686C12N2500/34G01N33/5044G01N2500/10
Inventor 姚卫峰张丽包贝华曹雨诞刘其南
Owner NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
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