摘要
目的 研究早期凋亡细胞的炎症抑制作用。方法 以紫外线照射诱导早期凋亡及坏死的Jurkat细胞 ,用流式细胞仪检测早期凋亡率。建立LPS诱导的小鼠炎症反应模型 ,分别向炎症部位滴注 1 0 0 μl灭菌磷酸缓冲液、坏死及早期凋亡Jurkat细胞。以HE染色切片病理学检查及激光共聚焦显微镜检查各处理组小鼠肺组织炎症反应程度 ;以双夹心ELISA、Westernblot、RT PCR等方法检测不同处理后小鼠炎症反应状态下细胞因子分泌和表达变化。结果 和注入灭菌磷酸缓冲液及坏死细胞相比 ,向炎症部位滴注早期凋亡细胞后 ,病理检查肺组织炎症反应程度明显减轻 ;激光共聚焦显微镜检查 ,肺间质CD3阳性荧光明显减少 ;肺组织Westernblot及RT PCR结果表明 ,注入外源性早期凋亡细胞后 ,LPS刺激下肺组织炎症性细胞因子MIP 2、TNF α的分泌及表达减少 ,抗炎性细胞因子TGF β1分泌增强 ;当TGF β1中和抗体和早期凋亡细胞共同滴入后 ,炎症性细胞因子MIP 2、TNF α的表达及分泌有所上调。腹腔灌洗液ELISA结果表明 ,早期凋亡细胞增强了LPS刺激的炎症性腹腔内抗炎性细胞因子TGF β1的分泌 ,抑制炎症性细胞因子MIP 2、TNF α的分泌。加入TGF β1中和抗体后逆转了早期凋亡细胞对MIP 2、TNF α分泌的抑制作用。结论 早期凋亡细胞通过增强炎症?
Objective To study the anti inflammation property of viable apoptotic cells. Methods To induce pulmonary inflammatory response LPS (0.5 mg/kg) was instilled into mice peritoneal. Pulmonary inflammatory response was determined by lung histology after 36 h of LPS stimulation. We applied ultraviolet irradiation to produce viable apoptotic cells and necrotic cells in vitro . 2×10 7 viable apoptotic or necrotic cells, were respectively instilled into the site of local inflammatory lung and peritonea after 36 h of LPS stimulation. PBS was instilled into LPS stimulated lung and peritonea respectively as control. Lung histology and confocal microscopy were applied to assay the lung inflammatory response. ELISA, Western blot and RT PCR were employed to determine the different effects of apoptotic and necrotic cells on cytokines expression and secretion in the inflammatory site. Results Confocal microscopy and HE section of lungs revealed significant reduction in parenchymal infiltrates and intra alveolar cells in the lungs instilled with apoptotic cells compared with those instilled with necrotic cells and PBS. Western blot and RT PCR revealed that 24 h after instillation of apoptotic cells endotracheally, the secretion and expression of pro inflammatory cytokines such as MIP 2 and TNF α were inhibited while the secretion of TGF β1 was increased when compared with instillation of necrotic cells and PBS endotracheally. When TGF β1 neutralization antibody was instilled together with apoptotic cells into LPS stimulated lung, the suppression effects on MIP 2 and TNF α secretion were abrogated. The supernatants of peritonea lavage was assessed by ELISA and got the same results as that obtained by Western blot. Conclusion Deliberate instillation of apoptotic cells into inflammatory site increased TGF β1 secretion and decreased MIP 2 and TNF α secretion which result in enhance resolution of inflammation.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2005年第3期208-213,共6页
Chinese Journal of Microbiology and Immunology
基金
国家重点基础研究发展计划 ( 973计划
NO .2 0 0 3CB5 1 5 5 0 0 )
国家自然科学基金资助项目 ( 30 371 35 8)
