CAJ | 학술논문

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、APJ mRNA表达的变化；Western blot法检测大鼠PMVECs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、APJ mRNA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,10-9~10-6 mol · 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,Western blot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。
목적탐토Apelin/APJ계통재지다당유도대서폐미혈관내피세포( PMVECs)손상중적변화,병연구Apelin적작용급궤제。방법채용식괴법배양대서PMVECs, VIII인자상관항원면역세포화학염색진행감정。새서람( MTT)비색법측정PMVECs활력；RT-PCR법검측Apelin、APJ mRNA표체적변화；Western blot법검측대서PMVECs중PCNA단백표체급Akt 적린산화수평。결과 LPS 재단시간내능구사Apelin、APJ mRNA표체수평정대상성상승( P<0．01),단수착작용시간연장,기인표체수도명현억제,저우대조조(P<0．05혹P<0．01),제시Apelin/APJ계통가능삼여료LPS유도적대서PMVECs손상。 MTT결과표명,10-9~10-6 mol · L-1적Apelin명현촉진료대서PMVECs증식( P<0．05혹P<0．01),차구유일정적농도화시간의뢰성。이차,Apelin환불동정도지개선료LPS유도적PMVECs세포손상( P<0．05혹P<0．01)。령외,Western blot결과현시,Apelin환명현역전료LPS유도적PCNA단백표체화Akt린산화수평적강저(P<0．05혹P<0．01)。결론 Apelin/APJ계통삼여료LPS유도적대서PMVECs손상。 Apelin대우유호폐미혈관내피세포공능,간예LPS유도적PMVECs손상기착중요작용,가능여기격활Akt린산화통로유관。
Aim To explore the changes of Apelin/APJ system in LPS-induced injury of rat pulmonary mi-crovascular 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 factorⅧrelated 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 promoted the 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.