Gene chip used for researching action of Alzheimer disease-controlling medicament

An Alzheimer's disease and gene chip technology, applied in the field of preparation of the chip, can solve problems such as adverse drug side effects, difficulty in obtaining satisfactory results, etc., achieve uniform concentration of synthetic products, and prevent amplification failure from affecting experiments and hybridization temperature. uniform effect

Inactive Publication Date: 2011-04-13
INST OF PHARMACOLOGY & TOXICOLOGY ACAD OF MILITARY MEDICAL SCI P L A
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, it is difficult to obtain satisfactory results by using traditional methods to study the pharmacological effects and mechanism of a

Method used

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  • Gene chip used for researching action of Alzheimer disease-controlling medicament
  • Gene chip used for researching action of Alzheimer disease-controlling medicament
  • Gene chip used for researching action of Alzheimer disease-controlling medicament

Examples

Experimental program
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Effect test

Embodiment 1

[0046] Example 1: Fabrication of gene chip for drug research on prevention and treatment of Alzheimer's disease

[0047] 1. Construction of DNA square matrix: 200 kinds of genes listed in Table 1 and negative controls (such as bacterial control probes), positive controls (such as eukaryotic housekeeping genes GAPDH, β-Actin) are arranged according to functional classification (or in random order). Each substrate is divided into 4 regions of equal size, and 200 molecules are spotted in each region, and each molecule is spotted twice.

[0048] 2. Probe design and synthesis: according to the three important parameters of the probe, namely sensitivity, specificity and the stability of forming a double strand with the target gene, the probe sequence is designed with a length of 60mer (or greater than 60mer). The oligonucleotides of 200 genes listed in Table 1 were synthesized by off-line solid-phase chemical method, the synthesized oligonucleotides were purified by PAGE method, an...

Embodiment 2

[0050] Example 2: Application of gene chip for drug research on prevention and treatment of Alzheimer's disease

[0051] 1. Total RNA extraction: TRIzol reagent, chloroform, isopropanol and 75% ethanol were used to extract the total RNA in the cells, tissues and patients' serum, cerebrospinal fluid and tissues before and after drug (or different drugs) treatment. The concentration and purity of total RNA were measured by ultraviolet spectrophotometer, and the integrity of total RNA was detected by formaldehyde-denatured agarose gel electrophoresis. last use Total RNA was purified by RNA Clean-up (740948.250, MN).

[0052] 2. Synthesis of cRNA

[0053] (1) Use the MessageAmpTM II-96aRNA Amplification kit (1819, Ambion) to reverse transcribe the total RNA into the first strand of cDNA;

[0054] (2) Use the MessageAmpTM II-96aRNA Amplification kit (1819, Ambion) to reverse transcribe the total RNA into the second strand of cDNA;

[0055] (3) use Extract II (740609.250, MN)...

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Abstract

The invention discloses a gene chip used for researching the action of an AD (Alzheimer Disease)-controlling medicament. An oligonucleotide gene probe array is fixed on the chip, wherein the oligonucleotide is an important biomolecule gene closely related to the occurrence and development process of the AD and relates to pathophysiological molecules, such as familial AD genes and susceptibility genes, amyloidosis, microtubule and cytoskeleton recombination, cynapse function, neurotransmission and ion channel, nerve regeneration, immunoregulation, immune response, signal transduction, chondriosome, protein metabolism, and the like. The chip of the invention can be used for researching pharmacological actions and action mechanisms of the AD-preventing and treating medicament containing a chemical medicament, a biotech drug, a single traditional Chinese medicine, a traditional Chinese medicine compound and an extract thereof, researching the sifting, the prescription, and the like of the AD-controlling medicament and researching the AD pathogenesis and carrying out auxiliary and differential diagnosis on a clinical case. The gene chip overcomes the defect of researching a multifactorial disease medicament by utilizing the method of a single target and has favorable application prospect.

Description

technical field [0001] The invention relates to a gene chip used for research on the pharmacological action and action mechanism of drugs for preventing and treating Alzheimer's disease, and also relates to a preparation method of the chip. Background technique [0002] Alzheimer's disease (AD), also known as senile dementia, is a serious neurodegenerative disease of human health. Clinically, it is characterized by progressive memory loss, cognitive dysfunction and behavioral abnormalities, and its pathological features are brain atrophy, senile plaques and neurofibrillary tangles in the brain, and neuron loss. [0003] Currently known, there are at least 32 types of dementia, among which AD accounts for as much as 50-60% of its incidence. According to the epidemiological survey, there are currently more than 25 million AD patients in the world. In 2008, the number of AD patients in the United States was estimated to reach 5.2 million, and 1 out of 8 people aged 65 and ove...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C40B40/06
Inventor 张永祥周文霞程肖蕊蒋宁杨胜张癸荣郑越黄璜孙琰钟乃凤王杉董蕾耿淼舒蕾赵月莹
Owner INST OF PHARMACOLOGY & TOXICOLOGY ACAD OF MILITARY MEDICAL SCI P L A
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