中华麻醉学杂志
中華痳醉學雜誌
중화마취학잡지
CHINESE JOURNAL OF ANESTHESIOLOGY
2010年
1期
75-78
,共4页
国斌%易斌%徐顺贵%鲁开智
國斌%易斌%徐順貴%魯開智
국빈%역빈%서순귀%로개지
内皮细胞%蛋白质丝氨酸苏氨酸激酶%肝肺综合征
內皮細胞%蛋白質絲氨痠囌氨痠激酶%肝肺綜閤徵
내피세포%단백질사안산소안산격매%간폐종합정
Endotheliai cells%Protein-serine-threonine kinases%Hepatopuimonary syndrome
目的 探讨肝肺综合征(HPS)大鼠血清对肺微血管内皮细胞(PMVECs)丝氨酸苏氨酸蛋白激酶(Akt)表达的影响.方法 健康3～4月龄SD大鼠30只,雌雄不拘,采用慢性胆管结扎法制备HPS模型.另取正常大鼠,原代培养、纯化及鉴定PMVECs.PMVECs接种于低糖DMEM培养基(10~6/,cm~2)或96孔培养板(200μl/孔),随机分为2组:对照组(C组)和HPS组,每组24皿或90孔,C组不予处理,HPS组加入HPS大鼠血清,血清终浓度为10%.于HPS大鼠血清中孵育24、48和72 h(T_(1～3))时分别采用RT-PCR法和Western blot法检测Akt_1 mRNA、Akt_2 mRNA和Akt_3 mRNA及其蛋白的表达,采用MTr法和~3H-TdR掺人法检测PMVECs增殖情况.结果 与C组比较,HPS组PMVECs增殖增强,Akt 蛋白及其mRNA表达水平上调(P<0.05);与T_1时比较,T_(2,3)时HPS组PMVECs增殖增强,Akt蛋白及其mRNA表达水平上调(P<0.05);与T_2时比较,T_3时HPS组PMVECs增殖增强,Akt蛋白及其mRNA表达水平上调(P<0.05).结论 Akt可能参与了HPS大鼠PMVECs增殖的调节.
目的 探討肝肺綜閤徵(HPS)大鼠血清對肺微血管內皮細胞(PMVECs)絲氨痠囌氨痠蛋白激酶(Akt)錶達的影響.方法 健康3～4月齡SD大鼠30隻,雌雄不拘,採用慢性膽管結扎法製備HPS模型.另取正常大鼠,原代培養、純化及鑒定PMVECs.PMVECs接種于低糖DMEM培養基(10~6/,cm~2)或96孔培養闆(200μl/孔),隨機分為2組:對照組(C組)和HPS組,每組24皿或90孔,C組不予處理,HPS組加入HPS大鼠血清,血清終濃度為10%.于HPS大鼠血清中孵育24、48和72 h(T_(1～3))時分彆採用RT-PCR法和Western blot法檢測Akt_1 mRNA、Akt_2 mRNA和Akt_3 mRNA及其蛋白的錶達,採用MTr法和~3H-TdR摻人法檢測PMVECs增殖情況.結果 與C組比較,HPS組PMVECs增殖增彊,Akt 蛋白及其mRNA錶達水平上調(P<0.05);與T_1時比較,T_(2,3)時HPS組PMVECs增殖增彊,Akt蛋白及其mRNA錶達水平上調(P<0.05);與T_2時比較,T_3時HPS組PMVECs增殖增彊,Akt蛋白及其mRNA錶達水平上調(P<0.05).結論 Akt可能參與瞭HPS大鼠PMVECs增殖的調節.
목적 탐토간폐종합정(HPS)대서혈청대폐미혈관내피세포(PMVECs)사안산소안산단백격매(Akt)표체적영향.방법 건강3～4월령SD대서30지,자웅불구,채용만성담관결찰법제비HPS모형.령취정상대서,원대배양、순화급감정PMVECs.PMVECs접충우저당DMEM배양기(10~6/,cm~2)혹96공배양판(200μl/공),수궤분위2조:대조조(C조)화HPS조,매조24명혹90공,C조불여처리,HPS조가입HPS대서혈청,혈청종농도위10%.우HPS대서혈청중부육24、48화72 h(T_(1～3))시분별채용RT-PCR법화Western blot법검측Akt_1 mRNA、Akt_2 mRNA화Akt_3 mRNA급기단백적표체,채용MTr법화~3H-TdR참인법검측PMVECs증식정황.결과 여C조비교,HPS조PMVECs증식증강,Akt 단백급기mRNA표체수평상조(P<0.05);여T_1시비교,T_(2,3)시HPS조PMVECs증식증강,Akt단백급기mRNA표체수평상조(P<0.05);여T_2시비교,T_3시HPS조PMVECs증식증강,Akt단백급기mRNA표체수평상조(P<0.05).결론 Akt가능삼여료HPS대서PMVECs증식적조절.
Objective To investigate the effect of the serum obtained from rat with hepatopuimonary syndrome (HPS) on Akt mRNA and protein expression in rat pulmonary microvascular endotheliai cells (PMVECs) and the role of Akt signaling pathway in the proliferation of PMVECs in the HPS. Methods Healthy 3-4-month-old SD rats of both sexes were used in this study. HPS was produced by chronic ligation of common bile duct according to the method described by Fallon. liver cirrhosis and pulmonary microvascular proliferation were verified by microscopic examination of the liver and lung tissue 2 weeks after bile duct ligation. Serum was obtained from blood taken from aorta of HPS rats. Primary PMVECs were cultured and randomly divided into 2 groups: control group and HPS group. In HPS group serum was added to cultured PMVECs (final concentration was 10%) and incubated. Akt mRNA and protein expression was determined at 24, 48 and 72 h of incubation by RT-PCR and Western blot. The proliferation of PMVECs was detected by MTT and ~3H-TdR. Results The proliferation of PMVECs was significantly enhanced and the expression of Akt mRNA and protein was significantly increased in HPS group as compared with control group. Conclusion The Akt signaling pathway might play an important role in proliferation of PMVECs in the HPS.
