Beijing da xue xue bao. Yi xue ban = Journal of Peking University. Health sciences · 2012
[Attenuation of experimental peritonitis and periodontitis in human chemokine-like receptor 1 (CMKLR1 or hChemR23) transgenic mice].
Аннотация на английскомСтатьяПоддержка исследования: вне правительства СШАНазвание и аннотация: Перевод пока не подготовлен
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Gao Li, Cyurko Robert, Van Dyke Thomas
Аннотация
OBJECTIVE: To prepare a transgenic mouse model with myeloid-selective expression of human chemokine-like receptor 1 (CMKLR1 or hChemR23) and investigate the in vivo inflammatory response of the particular transgenic mice in induced-peritonitis and periodontitis.
METHODS: The full-length hChemR23 cDNA and hCD11b promoters were cloned into pcDNA3 plasmid. Purified transgene (2.9 kb) was used to make transgenic mice at Boston University Transgenic/Knock out Mouse Core Facility. Copy numbers and mRNA level of hChemR23 transgene in the founders were verified by Custom TaqMan Gene Expression Real-time PCR assay. Hemizygous colonies were amplified by out-breeding hChemR23 transgenic males with wide-type FVB females. Transgenic mice were injected intraperitoneally with 1 mL of zymosan A solution (1 g/L in PBS). Peritoneal lavage cells stained with PE-conjugated anti-mouse F4/80 and FITC-conjugated anti-mouse Ly6G were analyzed by FACSort. Then 9-0 silk ligature was tied around the first molar to induce a periotontitis mouse model. The distance of cementoenamel junction to the alveolar bone crest (CEJ-ABC) was calculated to evaluate the alveolar bone loss. Data were analyzed with student's t-test and variance analysis.
RESULTS: One out of the four F1 founders carried the hChemR23 transgene. hChemR23 transgenic mice had reduced 56% fewer positive labeled polymorphonuclear neutrophil (PMNs) in peritonitis abdominal lavage fluid as compared with wide-type littermates. In a ligature-induced peirodontitis mouse model, hChemR23 transgenic mice showed decreased alveolar bone loss as compared with WT littermates (P<0.05).
CONCLUSION: The hChemR23 transgenic mice over-expressed the hChemR23 at mRNA level and had reduced inflammation in the acute peritonitis and ligature-induced periodontitis model.
METHODS: The full-length hChemR23 cDNA and hCD11b promoters were cloned into pcDNA3 plasmid. Purified transgene (2.9 kb) was used to make transgenic mice at Boston University Transgenic/Knock out Mouse Core Facility. Copy numbers and mRNA level of hChemR23 transgene in the founders were verified by Custom TaqMan Gene Expression Real-time PCR assay. Hemizygous colonies were amplified by out-breeding hChemR23 transgenic males with wide-type FVB females. Transgenic mice were injected intraperitoneally with 1 mL of zymosan A solution (1 g/L in PBS). Peritoneal lavage cells stained with PE-conjugated anti-mouse F4/80 and FITC-conjugated anti-mouse Ly6G were analyzed by FACSort. Then 9-0 silk ligature was tied around the first molar to induce a periotontitis mouse model. The distance of cementoenamel junction to the alveolar bone crest (CEJ-ABC) was calculated to evaluate the alveolar bone loss. Data were analyzed with student's t-test and variance analysis.
RESULTS: One out of the four F1 founders carried the hChemR23 transgene. hChemR23 transgenic mice had reduced 56% fewer positive labeled polymorphonuclear neutrophil (PMNs) in peritonitis abdominal lavage fluid as compared with wide-type littermates. In a ligature-induced peirodontitis mouse model, hChemR23 transgenic mice showed decreased alveolar bone loss as compared with WT littermates (P<0.05).
CONCLUSION: The hChemR23 transgenic mice over-expressed the hChemR23 at mRNA level and had reduced inflammation in the acute peritonitis and ligature-induced periodontitis model.