国际内分泌代谢杂志
國際內分泌代謝雜誌
국제내분비대사잡지
INTERNATIONAL JOURNAL OF ENDOCRINOLOGY AND METABOLISM
2013年
5期
293-297
,共5页
胰岛素样生长因子-1%结肠平滑肌细胞%细胞凋亡%丝裂原活化蛋白激酶通路
胰島素樣生長因子-1%結腸平滑肌細胞%細胞凋亡%絲裂原活化蛋白激酶通路
이도소양생장인자-1%결장평활기세포%세포조망%사렬원활화단백격매통로
Insulin-like growth factor-1%Colonic smooth muscle cells%Apoptosis%Mitogen-activated protein kinase pathway
目的 探讨胰岛素样生长因子-1(IGF-1)对大鼠结肠平滑肌细胞(SMCs)增殖、凋亡及丝裂原活化蛋白激酶(MAPK)信号转导通路的影响.方法 利用酶解法分离培养Sprague-Dawley大鼠结肠SMCs,进行免疫组化染色鉴定后,将其分为对照组、IGF-1组和IGF-1+ PD98059[细胞外信号调节激酶(ERK)抑制剂]组,分别采用噻唑蓝(MTT)法检测SMCs增殖,流式细胞术AnnexinV-FITC/PI检测SMCs凋亡,Western印迹检测磷酸化ERK、ERK、磷酸化p38MAPK、p38MAPK和磷酸化c-Jun氨基末端激酶(JNK)、JNK的表达.结果 分离培养的细胞经免疫组化鉴定为结肠SMCs,IGF-1组较对照组细胞增殖增强[(1.786 ±0.271)比(0.998±0.057),P＜0.01],凋亡率降低[(2.59±0.28)％比(20.68±2.48)％,P＜0.01],磷酸化ERK表达增强,磷酸化ERK/ERK比值升高[(42.71±3.74)％比(23.88±2.52％),P均＜0.01]；磷酸化p38MAPK、p38MAPK、磷酸化JNK、JNK表达无差异(P均＞0.05).IGF-1+ PD98059组较对照组细胞增殖下降[(0.154±0.021)比(0.998±0.057),P＜0.01],凋亡率升高[(84.31±7.54)％比(20.68±2.48)％,P＜0.01],磷酸化ERK表达减弱,磷酸化ERK/ERK比值降低[(10.47±1.22)％比(23.88±2.52)％,P均＜0.01].结论 IGF-1可能通过激活结肠SMCs MAPK通路中的ERK途径,促进细胞增殖,抑制凋亡,可能与p38MAPK途径和JNK途径无关.
目的 探討胰島素樣生長因子-1(IGF-1)對大鼠結腸平滑肌細胞(SMCs)增殖、凋亡及絲裂原活化蛋白激酶(MAPK)信號轉導通路的影響.方法 利用酶解法分離培養Sprague-Dawley大鼠結腸SMCs,進行免疫組化染色鑒定後,將其分為對照組、IGF-1組和IGF-1+ PD98059[細胞外信號調節激酶(ERK)抑製劑]組,分彆採用噻唑藍(MTT)法檢測SMCs增殖,流式細胞術AnnexinV-FITC/PI檢測SMCs凋亡,Western印跡檢測燐痠化ERK、ERK、燐痠化p38MAPK、p38MAPK和燐痠化c-Jun氨基末耑激酶(JNK)、JNK的錶達.結果 分離培養的細胞經免疫組化鑒定為結腸SMCs,IGF-1組較對照組細胞增殖增彊[(1.786 ±0.271)比(0.998±0.057),P＜0.01],凋亡率降低[(2.59±0.28)％比(20.68±2.48)％,P＜0.01],燐痠化ERK錶達增彊,燐痠化ERK/ERK比值升高[(42.71±3.74)％比(23.88±2.52％),P均＜0.01]；燐痠化p38MAPK、p38MAPK、燐痠化JNK、JNK錶達無差異(P均＞0.05).IGF-1+ PD98059組較對照組細胞增殖下降[(0.154±0.021)比(0.998±0.057),P＜0.01],凋亡率升高[(84.31±7.54)％比(20.68±2.48)％,P＜0.01],燐痠化ERK錶達減弱,燐痠化ERK/ERK比值降低[(10.47±1.22)％比(23.88±2.52)％,P均＜0.01].結論 IGF-1可能通過激活結腸SMCs MAPK通路中的ERK途徑,促進細胞增殖,抑製凋亡,可能與p38MAPK途徑和JNK途徑無關.
목적 탐토이도소양생장인자-1(IGF-1)대대서결장평활기세포(SMCs)증식、조망급사렬원활화단백격매(MAPK)신호전도통로적영향.방법 이용매해법분리배양Sprague-Dawley대서결장SMCs,진행면역조화염색감정후,장기분위대조조、IGF-1조화IGF-1+ PD98059[세포외신호조절격매(ERK)억제제]조,분별채용새서람(MTT)법검측SMCs증식,류식세포술AnnexinV-FITC/PI검측SMCs조망,Western인적검측린산화ERK、ERK、린산화p38MAPK、p38MAPK화린산화c-Jun안기말단격매(JNK)、JNK적표체.결과 분리배양적세포경면역조화감정위결장SMCs,IGF-1조교대조조세포증식증강[(1.786 ±0.271)비(0.998±0.057),P＜0.01],조망솔강저[(2.59±0.28)％비(20.68±2.48)％,P＜0.01],린산화ERK표체증강,린산화ERK/ERK비치승고[(42.71±3.74)％비(23.88±2.52％),P균＜0.01]；린산화p38MAPK、p38MAPK、린산화JNK、JNK표체무차이(P균＞0.05).IGF-1+ PD98059조교대조조세포증식하강[(0.154±0.021)비(0.998±0.057),P＜0.01],조망솔승고[(84.31±7.54)％비(20.68±2.48)％,P＜0.01],린산화ERK표체감약,린산화ERK/ERK비치강저[(10.47±1.22)％비(23.88±2.52)％,P균＜0.01].결론 IGF-1가능통과격활결장SMCs MAPK통로중적ERK도경,촉진세포증식,억제조망,가능여p38MAPK도경화JNK도경무관.
Objective To study the effects of insulin-like growth factor-1 (IGF-1) on the proliferation,apoptosis and mitogen-activated protein kinase (MAPK) signaling pathway of rat colonic smooth muscle cells (SMCs).Methods Colonic SMCs in Sprague Dawley were isolated and cultured by enzymatic method.After immunohistochemical staining for identification of SMCs,SMCs were divided into control group,IGF-1 group,and IGF-1 + PD98059[extracellular signal-regulated kinase(ERK) inhibitor] group.3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method was used for the detection of SMCs proliferation,flow cytometric Annexin V-FITC/PI for the detection of SMCs apoptosis,and Western blots for the detection of phosphorylated ERK,ERK,phosphorylated p38MAPK,p38MAPK and phosphorylated c-Jun N-terminal kinase(JNK),JNK expressions.Results The cultured cells were identified as colonic SMCs by immunohistochemical method.Compared with control group,cells proliferation of IGF-1 group was enhanced [(1.786 ± 0.271) vs.(0.998 ± 0.057),P ＜ 0.01],and apoptosis rate was reduced [(2.59 ± 0.28) ％ vs.(20.68 ±2.48) ％,P ＜ 0.01].Phosphorylated ERK expression in IGF-1 group was enhanced,and the ratio of phosphorylated ERK/ERK was increased [(42.71 ± 3.74) ％ vs.(23.88 ± 2.52) ％,all P ＜ 0.01].There were no differences for the expressions of phosphorylated p38MAPK,p38MAPK,phosphorylated JNK and JNK between IGF-1 group and control group (all P ＞ 0.05).Compared with control group,cells proliferation in IGF-1 ± PD98059 group was decreased significantly [(0.154 ± 0.021) vs.(0.998 ± 0.057),P ＜ 0.01],while apoptosis rate was increased [(84.31 ± 7.54) ％ vs.(20.68 ± 2.48) ％,P ＜ 0.01],phosphorylated ERK expression was reduced,and the ratio of phosphorylated ERK/ERK was decreased [(10.47 ± 1.22) ％ vs.(23.88 ± 2.52) ％,all P ＜ 0.01].Conclusion IGF-1 can promote the proliferation and inhibit apoptosis of colonic SMCs through activation of the ERK route of MAPK pathway,and p38MAPK and JNK routes may not be involved in this process.
