OBJECTIVE: To clone and study the polymorphism within interleukin-4 (IL-4) proximal promoter of asthmatic children. METHODS: The IL-4 proximal promoter segments were amplified and selected by polymerase chain reaction...OBJECTIVE: To clone and study the polymorphism within interleukin-4 (IL-4) proximal promoter of asthmatic children. METHODS: The IL-4 proximal promoter segments were amplified and selected by polymerase chain reaction (PCR) and single strand conformation polymorphism (SSCP) with genomic DNA from ten healthy children and forty patients with dominantly allergic familial histories as templates. The selected PCR segments were cloned into recombinant plasmids pIL-4-Jx2. The PCR inserts were sequenced by dideoxy chain termination method. RESULTS: Seven aberrant bands were found in SSCP analysis from forty asthmatic patients. The sequencing results showed that four variant sites were found within or adjacent to the known IL-4 regulatory element. A C to A transversion located at -229 position was just within the positive regulatory element-I (PRE-I) in one patient. A C to T transition adjacent to the negative regulatory element-II (NRE-II) and an extra G adjacent to TATA box were found in two patients. A five base nucleotide deletion was found near signal transducers and activators of transcription-6 responsive element (STAT-6 RE) in one patient. CONCLUSION: There were polymorphisms within the IL-4 proximal promoter of allergic asthmatic patients and these polymorphisms might result in aberrant expression of IL-4 gene and asthma.展开更多
Objective To explore the role of nuclear factor κB (NF κB) in the signal conduction of protein kinase C (PKC) regulated proliferation, apoptosis and expression of Th2 cytokines - interleukin 4 (IL 4) and interl...Objective To explore the role of nuclear factor κB (NF κB) in the signal conduction of protein kinase C (PKC) regulated proliferation, apoptosis and expression of Th2 cytokines - interleukin 4 (IL 4) and interleukin 5 (IL 5) of T lymphocytes in the bronchial alveolus lavage fluid (BALF) Methods T lymphocytes were isolated and purified from BALF of asthmatic guinea pigs in normal and asthmatic groups, and were stimulated with PKC agitator phorbol 12 myristate 13 acetate (PMA) and NF κB inhibitor pyrrolidine dithiocarbamate (PDTC), respectively The expressions of NF κB, IL 4 and IL 5 mRNA and protein, the proliferation and apoptosis of T lymphocytes were observed by immunohistochemistry, in situ hybridization, ELISA, MTT and TUNEL, respectively Results The activation of NF κB, proliferation response, and expression of IL 4 and IL 5 mRNA and protein in T lymphocytes stimulated by PMA were significantly higher than those of their blank control ( P 【0 01), while those indexes of T lymphocytes stimulated by PMA and PDTC simultaneously were significantly lower than those stimulated by PMA alone ( P 【0 01) The apoptotic index of T lymphocytes stimulated with PMA were significantly lower than that of their blank control ( P 【0 01), and the apoptotic index of asthmatic guinea pig T lymphocytes stimulated with PMA and PDTC simultaneously were significantly higher than that stimulated by PMA alone ( P 【0 01) The significant positive correlations were found between the activation of NF κB and the proliferation ( r =0 64, P 【0 001), and the expression of IL 4 and IL 5 mRNA and protein of T lymphocytes, respectively ( r =0 55-0 68, P 【0 001) There was also significant negative correlation between the activation of NF κB and apoptosis of T lymphocytes ( r =0 62, P 【0 001) Conclusions NF κB may participate in the signal conduction of PKC regulated proliferation, apoptosis and expression of IL 4 and IL 5 of T lymphocytes in asthma The activation of NF κB in PKC signal conduction pathway of T lymphocytes may play an important role in the pathogenesis of asthma展开更多
文摘OBJECTIVE: To clone and study the polymorphism within interleukin-4 (IL-4) proximal promoter of asthmatic children. METHODS: The IL-4 proximal promoter segments were amplified and selected by polymerase chain reaction (PCR) and single strand conformation polymorphism (SSCP) with genomic DNA from ten healthy children and forty patients with dominantly allergic familial histories as templates. The selected PCR segments were cloned into recombinant plasmids pIL-4-Jx2. The PCR inserts were sequenced by dideoxy chain termination method. RESULTS: Seven aberrant bands were found in SSCP analysis from forty asthmatic patients. The sequencing results showed that four variant sites were found within or adjacent to the known IL-4 regulatory element. A C to A transversion located at -229 position was just within the positive regulatory element-I (PRE-I) in one patient. A C to T transition adjacent to the negative regulatory element-II (NRE-II) and an extra G adjacent to TATA box were found in two patients. A five base nucleotide deletion was found near signal transducers and activators of transcription-6 responsive element (STAT-6 RE) in one patient. CONCLUSION: There were polymorphisms within the IL-4 proximal promoter of allergic asthmatic patients and these polymorphisms might result in aberrant expression of IL-4 gene and asthma.
文摘Objective To explore the role of nuclear factor κB (NF κB) in the signal conduction of protein kinase C (PKC) regulated proliferation, apoptosis and expression of Th2 cytokines - interleukin 4 (IL 4) and interleukin 5 (IL 5) of T lymphocytes in the bronchial alveolus lavage fluid (BALF) Methods T lymphocytes were isolated and purified from BALF of asthmatic guinea pigs in normal and asthmatic groups, and were stimulated with PKC agitator phorbol 12 myristate 13 acetate (PMA) and NF κB inhibitor pyrrolidine dithiocarbamate (PDTC), respectively The expressions of NF κB, IL 4 and IL 5 mRNA and protein, the proliferation and apoptosis of T lymphocytes were observed by immunohistochemistry, in situ hybridization, ELISA, MTT and TUNEL, respectively Results The activation of NF κB, proliferation response, and expression of IL 4 and IL 5 mRNA and protein in T lymphocytes stimulated by PMA were significantly higher than those of their blank control ( P 【0 01), while those indexes of T lymphocytes stimulated by PMA and PDTC simultaneously were significantly lower than those stimulated by PMA alone ( P 【0 01) The apoptotic index of T lymphocytes stimulated with PMA were significantly lower than that of their blank control ( P 【0 01), and the apoptotic index of asthmatic guinea pig T lymphocytes stimulated with PMA and PDTC simultaneously were significantly higher than that stimulated by PMA alone ( P 【0 01) The significant positive correlations were found between the activation of NF κB and the proliferation ( r =0 64, P 【0 001), and the expression of IL 4 and IL 5 mRNA and protein of T lymphocytes, respectively ( r =0 55-0 68, P 【0 001) There was also significant negative correlation between the activation of NF κB and apoptosis of T lymphocytes ( r =0 62, P 【0 001) Conclusions NF κB may participate in the signal conduction of PKC regulated proliferation, apoptosis and expression of IL 4 and IL 5 of T lymphocytes in asthma The activation of NF κB in PKC signal conduction pathway of T lymphocytes may play an important role in the pathogenesis of asthma
