摘要
目的．观察休克淋巴液对大鼠肺微血管内皮细胞（PMVECs）自由基及一氧化氮（NO）、肿瘤坏死因子-α（TNF—α）、白细胞介素-6（IL-6）表达的影响，进一步探讨休克淋巴液损伤PMVECs的机制。方法原代培养大鼠PMVECs至第3代进行研究。无菌条件下复制大鼠重症失血性休克模型（动脉压40mmHg维持90min，1mmHg-0．133kPa）。引流正常大鼠和休克大鼠肠系膜淋巴液及门静脉血，与PMVECs孵育6h，同时以胎牛血清（FBS）和无血清的DMEM培养液作为对照。逆转录-聚合酶链反应（RT—PCR）检测诱生型一氧化氮合酶（iNOs）、TNF—α和IL-6的mRNA表达；检测培养上清液中丙二醛（MDA）、NO、TNF—α和IL-6的含量。结果体积分数为4％终浓度的休克淋巴液作用6h后，PMVECs中iNOS、TNF-α和IL-6的mRNA表达以及培养上清液中MDA、NO、TNF—n和IL-6水平均显著高于空白对照组、正常淋巴液组、休克血浆组、正常血浆组和无血清对照组；且休克血浆作用PMVEC6h后的iN0s、TNF—α和IL-6的mRNA表达及培养上清液中N0水平均显著高于空白对照组、正常淋巴液组、正常血浆组和无血清对照组（P〈0．05或P〈0．01）。结论4％终浓度的休克淋巴液可致大鼠PMVECs中iNOS、TNF-α和IL-6的mRNA表达增强，促进自由基释放，从而诱导细胞损伤。
Objective To observe the effect of lymph collected during shock on free radical and expressions of nitric oxide （NO）, tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） mRNA of pulmonary micro-vascular endothelial cells （PMVECs） of rats in order to explore the mechanisms of damaging effect of lymph collected during shock to the PMVECs. Methods PMVECs were isolated and cultured, and used at passage 3. The model of serious hemorrhagic shock was reproduced by maintaining the arterial blood pressure of rats at 40 mm Hg （1 mm Hg：0. 133 kPa） for 90 minutes by exsanguination under aseptic condition. Mesentery lymph and portal vein blood were obtained from both shock rats and normal rats. PMVECs were respectively incubated in them for 6 hours, and at the same time, fetal bovine serum （FBS） and serum-free DMEM were used as culture media for comparison. The expressions of inducible nitric oxide synthase （iNOS）, TNF-α and IL-6 mRNA were detected by the method of reverse transcription- polymerase chain reaction （RT-PCR）, and the concents of malondialchehyche （MDA）, NO, TNF-α and IL-6 in culture supernatants were determined. Results After the PMVECs was treated by shock lymph at a final concentration of 4% for 6 hours, the expressions of iNOS, TNF-α and IL-6 mRNA in PMVECs and the contents of MDA, NO, TNF-α and IL-6 in culture supernatant fluids in shock lymph group were significantly increased compared with those of FBS group, normal lymph group, shock plasma group, normal plasma group and DMEM group. At the same time, the expressions of iNOS, TNF-α and IL-6 mRNA in PMVECs and the contents of NO in culture supernatant fluid of shock plasma group were significantly increased compared with those of FBS group, normal lymph group, normal plasma group and DMEM group （P〈0. 05 or P〈0.01）. Conclusion The results demonstrate that the shock lymph in final concentration of 4% could enhance the expressions of iNOS, TNF-α and IL-6 of PMVECs, reduce the free radical, and as a result, induce damage to PMVECs.
出处
《中国危重病急救医学》
CAS
CSCD
北大核心
2009年第5期266-269,共4页
Chinese Critical Care Medicine
基金
国家自然科学基金项目（30370561）
河北省自然科学基金项目（C2004000649）
关键词
休克
淋巴液
肺微血管内皮细胞
炎症介质
shock
lymph
pulmonary micro-vascular endothelial cell
inflammatory mediator
