摘要
目的 探讨Apelin/APJ系统在脂多糖诱导大鼠肺微血管内皮细胞（PMVECs）损伤中的变化,并研究Apelin的作用及机制。方法 采用植块法培养大鼠PMVECs,Ⅷ因子相关抗原免疫细胞化学染色进行鉴定。噻唑蓝（MTT）比色法测定PMVECs活力;RT-PCR法检测Apelin、APJmRNA表达的变化;Westernblot法检测大鼠PASMCs中PCNA蛋白表达及Akt的磷酸化水平。结果 LPS在短时间内能够使Apelin、APJmRNA表达水平呈代偿性上升（P〈0.01）,但随着作用时间延长,基因表达受到明显抑制,低于对照组（P〈0.05或P〈0.01）,提示Apelin/APJ系统可能参与了LPS诱导的大鼠PMVECs损伤。MTT结果表明,10-9~10-6mol·L-1的Apelin明显促进了大鼠PMVECs增殖（P〈0.05或P〈0.01）,且具有一定的浓度和时间依赖性。而且,Apelin还不同程度地改善了LPS诱导的PMVECs细胞损伤（P〈0.05或P〈0.01）。另外,Westernblot结果显示,Apelin还明显逆转了LPS诱导的PCNA蛋白表达和Akt磷酸化水平的降低（P〈0.05或P〈0.01）。结论 Apelin/APJ系统参与了LPS诱导的大鼠PMVECs损伤。Apelin对于维护肺微血管内皮细胞功能,干预LPS诱导的PMVECs损伤起着重要作用,可能与其激活Akt磷酸化通路有关。
Aim To explore the changes of Apelin/ APJ system in LPS-induced injury of rat pulmonary mi- erovascular endothelial cells（PMVECs） , and the effect and mechanism of Apelin. Methods PMVECs were cultured with the explant technique, and the identifica- tion of rat PMVECs was carried out by immunocyto- chemical staining of factor Ⅷ related antigen. MTT as- say was used to evaluate the viability of PMVECs. The mRNA expression of Apelin and APJ was detected by RT-PCR. The protein expression of PCNA and the phosphorylation of Akt was analyzed by Western blot. Results The mRNA expression of Apelin and APJ showed a compensatory increase after LPS treatment for a short period of time （P 〈0. 01 ）, but with the exten- sion of time, which was significantly inhibited, even lower than the control group （ P 〈 0. 05 or P 〈 0. 01 ）, suggesting that Apelin/ APJ system might be involved in LPS-induced PMVECs injury. MTT results showed that 10^-6 -10^-9 mol ·L-1 Apelin obviously promotedthe proliferation of rat PMVECs （P 〈 0.05 or P 〈 0. 01）, and with certain concentration and time de- pendence. Moreover, Apelin also improved the LPS- induced PMVECs injury in different degrees （P 〈 0.05 or P 〈 0. 01 ）. In addition, Western blot analysis showed that Apelin significantly reversed the decrease of the protein expression of PCNA and the Akt phos- phorylation level induced by LPS （ P 〈 0. 05 or P 〈 0. 01 ）. Conclusions The Apelin/APJ system is in- volved in LPS-induced PMVECs injury. Apelin plays an important role in protecting the pulmonary microvas- cular endothelial function and reversing the LPS-in- duced PMVECs injury, which might be related to the activation of Akt phosphorylation pathway.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2015年第8期1152-1158,共7页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 81273600)
河北省自然科学基金资助项目(No H2013206147)
关键词
APELIN
APJ
脂多糖
肺微血管内皮细胞
增殖
Akt
增殖细胞核抗原(PCNA)
Apelin
APJ
lipopolysaccharide （LPS）pulmonary microvascular endothelial ceils （PMVECs）proliferation
Akt
proliferating cell nuclear antigen（PCNA）
