体外培养肉鸡肺动脉内皮细胞的脂质过氧化损伤

Lipid peroxidatic injury of the cultured pulmonary artery endothelial cells of broiler in vitro

采用组织贴块法培养肉鸡肺动脉内皮细胞(PAEC),并依据细胞形态学特点和凝血因子Ⅷ相关抗原免疫组化染色结果对所培养的细胞进行鉴定。以黄嘌呤/黄嘌呤氧化酶(X/XO)系统作为氧自由基发生系统建立PAEC氧化损伤模型。结果表明:肉鸡PAEC在体外能够存活并传5-6代。形态学观察和Ⅷ因子抗原免疫组化染色检查证明培养的细胞符合肺动脉内皮细胞的特点。氧自由基能够明显损伤PAEC,X/XO系统作用下细胞生长被抑制,甚至导致细胞死亡,并且这种抑制作用对XO有剂量依赖性。与正常对照组相比,损伤组PAEC培养液中脂质过氧化产物丙二醛(MDA)含量升高,提示氧自由基促进了PAEC的脂质过氧化。这说明氧自由基对肉鸡肺动脉内皮细胞具有损伤作用,可能在肉鸡肺动脉高压综合征的发病过程中起重要作用。

Pulmonary artery endothelial cells （PAEC） of broiler were isolated by the method of tissue explantation. Identifications of the cultured cells were performed with the morphological feature and immunocytochemical stain using specific antiserum against factor Ⅷ related antigen. Xanthine/xanthine oxidase （X/XO） served as the oxygen free radical （OFR） generating system. In vitro model of oxidative injury of PAEC was established based on the X/XO system. The results showed that PAEC could survive in vitro and subculture for 5 - 6 generations. Morphological and immunocytochemical observations of cultured cells demonstrated specific characteristics of endothelial cells. PAECs were severely damaged by OFR. The viability of cells was inhibited by X/XO system, and dose-dependent decrease in cell viability was found with increasing XO dosages. Some cells even died. The concentration of malondialdehyde （ MDA, a product of lipid peroxidation） was increased in the culture media treated with X/XO system, suggesting that OFR promoted the lipid peroxidation of PAEC. These results suggest that OFR might have an injury effect on the endothelium of pulmonary artery in broiler and may play an important role in the mechanism of pulmonary hypertension syndrome.