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A technology of inhibitors and cells, applied in the field of drugs for lung injury in mammals, preparation of drugs for preventing and/or treating avian influenza in mammals, and can solve problems such as unclear specific mechanisms
Inactive Publication Date: 2010-10-27
THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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Problems solved by technology
However, the specific mechanism of the interaction between nanomaterials and cells is still unclear, and the mechanism of nanomaterials causing lung injury by inducing cell autophagy has not been reported yet.
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Embodiment 1
[0127] Codon optimization and humanization gene optimization methods are used to replace the rarely used codons in the expression host of the H5 protein gene coding with frequently used codons. Find the amino acid sequence corresponding to the wild-type H5N1 virus H5 protein (A / Thailand / 4(SP-528) / 2004(H5N1)) through NCBI, and replace the corresponding codons with codons with a high proportion of expression host usage frequency , so as to obtain several optimized H5 protein gene sequences, use DNAMAN software to remove the sequences that may form complex secondary structures, select the optimized H5 genes, remove the transmembrane sequence, and perform gene synthesis in Beijing Qingke Biological Co., Ltd. as Construct the target gene of the plasmid.
[0128] The synthetic sequence is shown as SEQ ID NO:1.
Embodiment 2
[0130] Plasmid construction and detection (see the third edition of > for the experimental method).
[0131] Carrier: Peak13 CD5L TEV human IgG (preserved in our laboratory).
[0132] Insert: Optimized H5 gene with transmembrane removed.
[0133] After the enzyme digestion, electrophoresis, purification, ligation, and transformation processes shown in molecular cloning, the insert fragments were identified, and finally the correct plasmids detected by enzyme digestion (Nhe I / BamH I) were sequenced (Beijing Qingke Biological Co., Ltd. ) for further confirmation.
[0134] The experimental results are attached figure 1 As shown, the three lanes 1, 2, and 3 are λ-Hind III Marker, Peak13 CD5LH5 TEV human IgG, and D2000 Marker. λ-Hind III Marker from small to large (from bottom to top) is 564bp (difficult to distinguish from the picture), 2027bp, 2322bp, 4361bp, 6557bp, 9416bp, 23130bp; D2000Marker from small to large (from bottom to top) The order is 100bp, 250bp, 500bp, 750bp,...
Embodiment 3
[0135] Example 3 Large-scale extraction of plasmids
[0136] A large number of recombined plasmids were extracted by CsCl density gradient centrifugation (see the third edition of > for the experimental method).
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Abstract
The invention relates to a use of an autophagy (B-cell apoptosis) inhibitor. The inventor finds that the autophagy (B-cell apoptosis) inhibitor can be used for preventing and / or curing bird flu, and particularly has surprising effect on bird flu prevention. The inventor additionally finds that the autophagy (B-cell apoptosis) inhibitor can also be used for preventing and / or curing lung injuries caused or deteriorated by nano materials. The autophagy (B-cell apoptosis) inhibitor comprises 3-methyladenine, SB203580, LY294002 or wortmannin. The effect of the autophagy (B-cell apoptosis) inhibitor is very obvious, the occurrence of autophagy can be obviously inhibited, the cell deaths, especially the deaths of lung epithelial cells of mammals are greatly reduced, the injury degree of organs, especially the injury degree of the lung is reduced, and thereby the bird flu can be prevented and cured; and the autophagy (B-cell apoptosis) inhibitor can prevent and cure the lung injuries caused or deteriorated by the nano materials.
Description
technical field [0001] The present invention relates to the use of inhibitors of autophagy (type II cell apoptosis). Specifically, the present invention relates to inhibitors of autophagy (type II cell apoptosis) such as 3-methyladenine, SB203580, LY294002 or wortmannin in the preparation of medicaments for preventing and / or treating avian influenza in mammals the use of. The present invention also relates to the use of the inhibitor of cell autophagy (type II cell apoptosis) in the preparation of medicines for preventing and / or treating mammalian lung damage caused by nanomaterials. Background technique [0002] Influenza virus is the pathogenic virus that causes influenza. It is an RNA virus that can cause acute respiratory infectious diseases and lead to various complications such as myocarditis, pneumonia, and bronchitis. Because influenza viruses are highly contagious, epidemics, and even worldwide pandemics, can easily occur. The 1917-1919 flu outbreak in Europe kil...
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