国际呼吸杂志
國際呼吸雜誌
국제호흡잡지
INTERNATIONAL JOURNAL OF RESPIRATION
2008年
17期
1038-1041
,共4页
转化生长因子α%气道平滑肌细胞%表皮生长因子受体
轉化生長因子α%氣道平滑肌細胞%錶皮生長因子受體
전화생장인자α%기도평활기세포%표피생장인자수체
Transforming growth factor alpha%Airway smooth muscle cells%Epidermal growth factor receptor
目的 探讨转化生长因子α(transforming growth factog alphg,TGF-α)对小鼠气道平滑肌细胞(airway smooth muscle cells,ASMC)促增殖作用及其机制.方法 用四唑盐(MTT)比色法和3H-TdR掺人法测定加入TGF-α后小鼠ASMC的增殖情况.本实验采用3H-TdR掺入法和MTT比色法观察选择性表皮生长因子受体(epidermal growth factor receptor,EGFR)酪氨酸激酶抑制剂(AG1478)、EGFR中和抗体225(225mAb)、MEK抑制剂(U0126)、PI-3K抑制剂(Wonmannin)对加入TGF-α后促ASMC增殖的影响.通过Western Blot方法测定加入TGF-α及加用AG1478、225mAb后ASMC磷酸化EGFR蛋白表达.结果 用MTT比色法和3H-TdR掺入法测定ASMC培养液中加入TGF-α后ASMC增殖情况比对照组明显增加.加入AG1478、225mAb、U0126、Wortmannin可抑制TGF-α促ASMC增殖作用(P＜0.01).经Western Blot检测,TGF-α引起ASMC磷酸化EGFR蛋白表达增高.AG1478、225mAb抑制TGF-α所致ASMC磷酸化EGFR蛋白表达的增加(P＜0.01).结论 TGF-α激活EGFR为磷酸化EGFR,从而通过:①ras-raf-MEK-erk/MAPK途径;②PI3K-PKC-IKK途径;促进体外培养的小鼠ASMC的增殖.
目的 探討轉化生長因子α(transforming growth factog alphg,TGF-α)對小鼠氣道平滑肌細胞(airway smooth muscle cells,ASMC)促增殖作用及其機製.方法 用四唑鹽(MTT)比色法和3H-TdR摻人法測定加入TGF-α後小鼠ASMC的增殖情況.本實驗採用3H-TdR摻入法和MTT比色法觀察選擇性錶皮生長因子受體(epidermal growth factor receptor,EGFR)酪氨痠激酶抑製劑(AG1478)、EGFR中和抗體225(225mAb)、MEK抑製劑(U0126)、PI-3K抑製劑(Wonmannin)對加入TGF-α後促ASMC增殖的影響.通過Western Blot方法測定加入TGF-α及加用AG1478、225mAb後ASMC燐痠化EGFR蛋白錶達.結果 用MTT比色法和3H-TdR摻入法測定ASMC培養液中加入TGF-α後ASMC增殖情況比對照組明顯增加.加入AG1478、225mAb、U0126、Wortmannin可抑製TGF-α促ASMC增殖作用(P＜0.01).經Western Blot檢測,TGF-α引起ASMC燐痠化EGFR蛋白錶達增高.AG1478、225mAb抑製TGF-α所緻ASMC燐痠化EGFR蛋白錶達的增加(P＜0.01).結論 TGF-α激活EGFR為燐痠化EGFR,從而通過:①ras-raf-MEK-erk/MAPK途徑;②PI3K-PKC-IKK途徑;促進體外培養的小鼠ASMC的增殖.
목적 탐토전화생장인자α(transforming growth factog alphg,TGF-α)대소서기도평활기세포(airway smooth muscle cells,ASMC)촉증식작용급기궤제.방법 용사서염(MTT)비색법화3H-TdR참인법측정가입TGF-α후소서ASMC적증식정황.본실험채용3H-TdR참입법화MTT비색법관찰선택성표피생장인자수체(epidermal growth factor receptor,EGFR)락안산격매억제제(AG1478)、EGFR중화항체225(225mAb)、MEK억제제(U0126)、PI-3K억제제(Wonmannin)대가입TGF-α후촉ASMC증식적영향.통과Western Blot방법측정가입TGF-α급가용AG1478、225mAb후ASMC린산화EGFR단백표체.결과 용MTT비색법화3H-TdR참입법측정ASMC배양액중가입TGF-α후ASMC증식정황비대조조명현증가.가입AG1478、225mAb、U0126、Wortmannin가억제TGF-α촉ASMC증식작용(P＜0.01).경Western Blot검측,TGF-α인기ASMC린산화EGFR단백표체증고.AG1478、225mAb억제TGF-α소치ASMC린산화EGFR단백표체적증가(P＜0.01).결론 TGF-α격활EGFR위린산화EGFR,종이통과:①ras-raf-MEK-erk/MAPK도경;②PI3K-PKC-IKK도경;촉진체외배양적소서ASMC적증식.
Objective To study the role of transforming growth factor alpha (TGF-α) on the proliferation of airway smooth muscle cells (AMSC) and the mechanism. Methods MTT and 3H-TdR incorporation assays were used to evaluate the proliferation of AMSC after TGF-α were added. The effect of AG1478,22SmAb, U0126 and Wortmannin on TGF-α inducing proliferation of ASMC were assessed by MTT assay and 3H-TdR incorporation assays. Phospho-EGF receptor protein in ASMC was detected by Western blot analysis with TGF-α,AG1478 or 225mAb. Results The OD value of MTT assay and the CPM value of 3H-TdR incorporation assay of ASMC stimulated with TGF-α were significantly higher than those of control group (P＜0.01). AG1478, 225mAb, U0126 and Wortmannin inhibited TGF-α inducing proliferation of ASMC (P＜0.01). By Western blot analysis, the expression of phospho-EGF receptor proteinin ASMC stimulated with TGF-α(P ＜ 0.05). AG1478,225mAb inhibited the increase (P＜ 0.01).Conclusions TGF-α induces proliferation of ASMC via two pathways. One is Ras-raf-MEK-erk pathway, the other is PI3K-PKC-IKK pathway.
