安徽医科大学学报
安徽醫科大學學報
안휘의과대학학보
ACTA UNIVERSITY MEDICINALIS ANHUI
2014年
6期
726-729,730
,共5页
牛蒡子苷元%细胞增殖%IL-2%IFN-γ%mTOR
牛蒡子苷元%細胞增殖%IL-2%IFN-γ%mTOR
우방자감원%세포증식%IL-2%IFN-γ%mTOR
Arctigenin%cell proliferation%IL-2%IFN-γ%mTOR
目的：研究牛蒡子苷元(ATG)对刀豆蛋白A(ConA)诱导的小鼠脾细胞增殖和相关细胞因子分泌的影响及其可能的分子机制。方法 MTT法检测ATG对小鼠脾细胞的毒性；H3-胸腺嘧啶核苷掺入法检测ATG对小鼠脾细胞的增殖抑制水平；ELISA法检测细胞因子干扰素γ( IFN-γ)和白介素2(IL-2)的分泌水平；Western blot法检测mTOR通路相关蛋白(mTOR、P70S6K、Akt、AMPK、Raptor)的表达及磷酸化水平。结果 ATG对小鼠脾细胞无显著毒性；ATG显著抑制了ConA诱导的小鼠脾细胞增殖；ATG显著减少了小鼠脾细胞上清液中淋巴因子IFN-γ和IL-2的含量，且具有浓度依赖性；ATG显著降低了mTOR和P70S6K的磷酸化水平，增加了上游AMPK和Raptor的磷酸化，而Akt的磷酸化水平没有明显变化。结论 ATG可显著抑制ConA诱导的小鼠脾细胞增殖并减少 IFN-γ、IL-2的分泌，该作用可能与其增强AMPK、Raptor 的磷酸化，抑制 mTOR、P70S6K 的磷酸化有关。
目的：研究牛蒡子苷元(ATG)對刀豆蛋白A(ConA)誘導的小鼠脾細胞增殖和相關細胞因子分泌的影響及其可能的分子機製。方法 MTT法檢測ATG對小鼠脾細胞的毒性；H3-胸腺嘧啶覈苷摻入法檢測ATG對小鼠脾細胞的增殖抑製水平；ELISA法檢測細胞因子榦擾素γ( IFN-γ)和白介素2(IL-2)的分泌水平；Western blot法檢測mTOR通路相關蛋白(mTOR、P70S6K、Akt、AMPK、Raptor)的錶達及燐痠化水平。結果 ATG對小鼠脾細胞無顯著毒性；ATG顯著抑製瞭ConA誘導的小鼠脾細胞增殖；ATG顯著減少瞭小鼠脾細胞上清液中淋巴因子IFN-γ和IL-2的含量，且具有濃度依賴性；ATG顯著降低瞭mTOR和P70S6K的燐痠化水平，增加瞭上遊AMPK和Raptor的燐痠化，而Akt的燐痠化水平沒有明顯變化。結論 ATG可顯著抑製ConA誘導的小鼠脾細胞增殖併減少 IFN-γ、IL-2的分泌，該作用可能與其增彊AMPK、Raptor 的燐痠化，抑製 mTOR、P70S6K 的燐痠化有關。
목적：연구우방자감원(ATG)대도두단백A(ConA)유도적소서비세포증식화상관세포인자분비적영향급기가능적분자궤제。방법 MTT법검측ATG대소서비세포적독성；H3-흉선밀정핵감참입법검측ATG대소서비세포적증식억제수평；ELISA법검측세포인자간우소γ( IFN-γ)화백개소2(IL-2)적분비수평；Western blot법검측mTOR통로상관단백(mTOR、P70S6K、Akt、AMPK、Raptor)적표체급린산화수평。결과 ATG대소서비세포무현저독성；ATG현저억제료ConA유도적소서비세포증식；ATG현저감소료소서비세포상청액중림파인자IFN-γ화IL-2적함량，차구유농도의뢰성；ATG현저강저료mTOR화P70S6K적린산화수평，증가료상유AMPK화Raptor적린산화，이Akt적린산화수평몰유명현변화。결론 ATG가현저억제ConA유도적소서비세포증식병감소 IFN-γ、IL-2적분비，해작용가능여기증강AMPK、Raptor 적린산화，억제 mTOR、P70S6K 적린산화유관。
Objective To investigate the effects of Arctigenin ( ATG ) on concanavalin ( ConA )-stimulated cell proliferation and cytokine secretion in mouse spleen cells, and its possible mechanism. Methods The toxicity of ATG on mouse spleen cells was determined by MTT assay. The inhibition of proliferation was investigated by tritiat-ed thymidine incorporation method. Secreted cytokines (IFN-γand IL-2) were analyzed by ELISA. The associated proteins and phosphorylation levels of mTOR pathway ( mTOR/P70 S6 K/Akt/AMPK/Raptor ) were detected by Western blot. Results ATG had no significant toxicity to mouse spleen cells. ATG significantly inhibited mouse primary spleen cells proliferation induced by ConA. ATG suppressed IL-2 and IFN-γ production of mouse spleen cells in a concentration-dependent manner. ATG remarkably suppressed the phosphorylation of mTOR and P70S6K, and enhanced the phosphorylation of upstream AMPK and Raptor, while the phosphorylation of Akt did not change significantly. Conclusion ATG markedly suppresses the proliferation of mouse spleen stimulated by ConA cells and secretion of IFN-γand IL-2 , which may be correlated to the abilities of enhancing the phosphoryla-tion of AMPK and Raptor, inhibiting the phosphorylation of mTOR and P70S6K.
