ABSTRACT
The pathogenesis of allergic rhinitis(AR)is extremely complex.In recent years,a variety of allergens and other complexes have been developed to induce a series of signal transduction mechanisms by activating mast cells.Intracellular media release(mast cells,MCs)play an important role in the pathogenesis of AR.In this paper,we reviewed the progress of mast cells in the pathogenesis of allergic rhinitis in recent years in order to further understand its role in the pathogenesis of allergic rhinitis and provide new ideas on the therapeutic target for allergic rhinitis.
Subject(s)
Humans , Allergens , Cell Count , Mast Cells , Rhinitis, Allergic , Allergy and Immunology , Signal TransductionABSTRACT
OBJECTIVE@#To observe the clinical manifestations of allergic rhinitis mice and the expression changes of the eosinophils CCR3 and the granule protein mRNA in the bone marrow, peripheral blood and nasal lavage fluid.@*METHODS@#Twenty-four BALB/c mice were randomly divided into the control group, PBS therapy group, siRNA therapy group and the CCR3 siRNA therapy group (n=6). Allergic rhinitis model were sensitized and stimulated by ovalbunfin, and CCR3 siRNA therapy group were administered with CCR3 transnasally before stimulated. The levels of the eosinophils CCR3, MBP, ECP and EPO in bone marrow, peripheral blood and nasal lavage fluid were detected by RT-PCR.@*RESULTS@#Compared to the control group and CCR3 siRNA therapy group, the nasal mucosa of the PBS therapy group and siRNA therapy group developed epithalaxy, goblet cells hyperplasia, squamous epithelium metaplasia, epithelium necrosis, lamina propria and submucosa gland hyperplasia, vasodilatation, tissue edema, and the characterized eosinophil infiltration. RT-PCR indicated that the CCR3 mRNA, MBP, ECP and EPO expression in bone marrow, peripheral blood and nasal lavage fluid of the CCR3 siRNA therapy group was lower than the PBS therapy group and siRNA therapy group (P<0.05).@*CONCLUSIONS@#The RNA interference therapy to CCR3 by local administration pernasal can suppress the process of the development, migration and invasion of the allergic rhinitis eosinophil, thus can reduce the effect of eosinophils and then reduce the inflammation effect of the allergic rhinitis. It may be a new treatment for respiratory tract allergic inflammation.
Subject(s)
Animals , Male , Mice , Behavior, Animal , Bone Marrow , Chemistry , Disease Models, Animal , Eosinophil Granule Proteins , Genetics , Metabolism , Eosinophils , Metabolism , Physiology , Mice, Inbred BALB C , Nasal Mucosa , Chemistry , Cell Biology , RNA, Small Interfering , Genetics , Random Allocation , Receptors, CCR3 , Genetics , Metabolism , Rhinitis, Allergic , Rhinitis, Allergic, Perennial , Genetics , TherapeuticsABSTRACT
<p><b>OBJECTIVE</b>Through construction of a lentiviral expression vector of chemokine receptor 3 (CCR3)RNA interference (RNAi) of mouse, to further study the function of CCR3 gene on eosinophils.</p><p><b>METHODS</b>Focused on the CCR3 gene sequences, RNAi target sequences were designed, then the target sequences of Oligo DNA were synthesized and annealed to double stranded DNA, which was subsequently connected to pLVX-shRNA2-m vector digested by MluI, SacI, EcoRI, HindIII, BamHI and Xho I, short hairpin RNA lentiviral vectors were constructed. Short hairpin RNA lentiviral vectors were constructed. 293T cells and eosinophils were transfected by shRNA lentiviral vector, and virus titer was determined. The expression of the CCR3 gene in eosinophils was identified by quantitative-PCR.</p><p><b>RESULTS</b>The lentiviral vector of shRNA-mCCR3-oligonucleotide chain was inserted correctly. Infection efficiency of 293T cells observed under fluorescence microscope was more than 90%, the virus titer was 4×10(8) TU/ml. CCR3 interference rate was 86.7%.</p><p><b>CONCLUSION</b>A lentiviral vector of CCR3-gene RNAi was constructed successfully by the genetic engineering technology, and it provides a condition for further research in vitro and vivo.</p>
Subject(s)
Animals , Mice , Base Sequence , DNA , Eosinophils , Metabolism , Genetic Vectors , Lentivirus , Genetics , Polymerase Chain Reaction , RNA Interference , RNA, Small Interfering , Receptors, CCR3 , Genetics , Metabolism , TransfectionABSTRACT
The regulatory region and the signal peptide sequence of the sacB gene has been amplified by PCR using Bacillus subtilis chromosomal DNA as template, and an inducible secretion vector has been developed based on this sequence, which was ligated with Bacillus subtilis alkaline proteinase gene. Transform Bacillus subtilis DB403 with this vector, and the expression of the inserted Bacillus subtilis alkaline proteinase gene can be induced by addition of sucrose into the medium.
ABSTRACT
The degQ gene, amplified from Bacillus subtilis by PCR, was cloned to pUBS (sucrose induced secretion vector). After transformed into DB403, recombination named DB403(pUBSD) was formed. The results of the fermentation showed that degQ gene enhanced the expression of B. subtilis fibrin enzyme. The activity of the enzyme was increased to 2.2 times as the original one. In this article, the effects of different conditions, such as different kinds of sugar, different concentration of sucrose and different induced time were also be investigated and compared.