Zhang Ning, Xu Yong-jian, Zhang Zhen-xiang, Xiong Wei-ning
Department of Respiratory Medicine, Tongji Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Zhonghua Jie He He Hu Xi Za Zhi. 2004 Jan;27(1):36-40.
To investigate the effect of nuclear factor-kappaB (NF-kappaB) decoy oligodeoxynucleotide (ODN) on proliferation, apoptosis and cytokine synthesis of T lymphocytes from asthmatic patients.
T lymphocytes were divided into four groups, a normal control group (A group), an asthma control group (B group), a NF-kappaB cis decoy ODN group (B(1) group) and a scrambled ODN group (B(2) group). B(1) and B(2) groups were transfected by cationic lipofectamine to the latter two groups respectively. The proliferation of T lymphocytes was measured by MTT and the apoptosis of them was measured by flow cytometry. The expression levels of interleukin 5 (IL-5) mRNA and protein were detected with cell hybridization in situ and enzyme-linked immunosorbent assay (ELISA). The expression levels of inducible nitric oxide synthase (iNOS) were measured by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot.
The difference of proliferation rate between B(1) group (0.220 +/- 0.020) and B group (0.340 +/- 0.030) was significant (P < 0.05). The difference of apoptosis rate between B(1) group (10.8 +/- 1.3) and B group (8.1 +/- 1.2) was also significant (P < 0.05). The expression levels of IL-5 mRNA and protein in B(1) group (21 +/- 4, 24 +/- 4) were significantly different from those in B group (33 +/- 4, 54 +/- 10, P < 0.05). The differences of iNOS mRNA and protein levels between B(1) group (0.33 +/- 0.05, 782 +/- 117) and B group (0.75 +/- 0.13, 1185 +/- 230) were significant (P < 0.05). However, these indices in B(2) group showed no difference to those in B group (P > 0.05).
NF-kappaB decoy ODN can reduce the abnormally increased proliferation of T lymphocytes in asthma and increase the abnormally decreased apoptosis of these cells, while decrease the abnormally increased levels of cytokine and enzyme in asthmatic T lymphocytes. These may be the mechanisms underlying its potential therapeutic effects in asthma.
探讨核因子-κB（NF-κB）诱饵寡脱氧核苷酸（ODN）对哮喘患者T淋巴细胞增殖、凋亡及细胞因子合成的影响。
将T淋巴细胞分为四组，正常对照组（A组）、哮喘对照组（B组）、NF-κB顺式诱饵ODN组（B₁组）和乱序ODN组（B₂组）。分别用阳离子脂质体将B₁组和B₂组转染至后两组。采用MTT法检测T淋巴细胞的增殖情况，用流式细胞术检测其凋亡情况。采用原位细胞杂交和酶联免疫吸附测定（ELISA）检测白细胞介素5（IL-5）mRNA和蛋白的表达水平。采用逆转录-聚合酶链反应（RT-PCR）和蛋白质免疫印迹法检测诱导型一氧化氮合酶（iNOS）的表达水平。
B₁组（0.220±0.020）与B组（0.340±0.030）的增殖率差异有统计学意义（P<0.05）。B₁组（10.8±1.3）与B组（8.1±1.2）的凋亡率差异也有统计学意义（P<0.05）。B₁组IL-5 mRNA和蛋白的表达水平（21±4，24±4）与B组（33±4，54±10）相比差异有统计学意义（P<0.05）。B₁组（0.33±0.05，782±117）与B组（0.75±0.13，1185±230）的iNOS mRNA和蛋白水平差异有统计学意义（P<0.05）。然而，B₂组的这些指标与B组相比无差异（P>0.05）。
NF-κB诱饵ODN可降低哮喘患者T淋巴细胞异常增高的增殖水平，增加其异常降低的凋亡水平，同时降低哮喘患者T淋巴细胞中异常增高的细胞因子和酶的水平。这些可能是其对哮喘潜在治疗作用的机制。
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