本文選題：SP + c-Jun氨基末端激酶通路　； 參考：《中山大學(xué)學(xué)報(醫(yī)學(xué)科學(xué)版)》2014年02期

【摘要】：【目的】研究抑制UVA激活的皮膚成纖維細(xì)胞JNK通路的最佳SP600125濃度,為研究JNK信號通路在光老化中的作用機(jī)制奠定基礎(chǔ)�！痉椒ā吭囵B(yǎng)取自兒童包皮的皮膚成纖維細(xì)胞。用CCK-8法結(jié)合熒光倒置顯微鏡研究不同劑量UVA和不同濃度SP600125及兩者聯(lián)合對皮膚成纖維細(xì)胞活性和形態(tài)的影響,以選擇非細(xì)胞毒性的UVA劑量和SP600125濃度。再用Western-blot檢測UVA輻照后不同時間點(diǎn)皮膚成纖維細(xì)胞中磷酸化c-Jun(P-c-Jun)表達(dá)以確定其被誘導(dǎo)表達(dá)的時間點(diǎn)。最后用Western-blot檢測不同濃度SP600125對UVA上調(diào)的P-c-Jun表達(dá)的抑制。【結(jié)果】UVA和SP600125分別輻照細(xì)胞和孵育細(xì)胞時,UVA≤10 J/cm2和SP600125≤2μmol/L均能保持細(xì)胞活性在90%以上。10 J/cm2的UVA輻照不同濃度SP600125孵育的細(xì)胞時,≤800 nmol/L的SP600125孵育的細(xì)胞活性在90%以上;1μmol/L的SP600125使細(xì)胞活性明顯下降至67.1%,鏡下見部分細(xì)胞皺縮、碎裂、死亡;2μmol/L的SP600125使細(xì)胞活性進(jìn)一步下降至7.6%,鏡下見絕大部分細(xì)胞變形、碎裂、死亡。Western-blot結(jié)果顯示P-c-Jun在UVA輻照后0.75 h、1.5 h均較0 h升高,3 h后恢復(fù)至0 h水平。200~800 nmol/L SP600125呈劑量依賴性地抑制UVA上調(diào)的P-c-Jun表達(dá),差異有統(tǒng)計(jì)學(xué)意義�！窘Y(jié)論】低于常用濃度的200~800 nmol/L SP600125即可抑制UVA激活的皮膚成纖維細(xì)胞的JNK通路。JNK通路可能在UVA輻照的皮膚成纖維細(xì)胞中起抗凋亡作用。
[Abstract]:[objective] to study the optimal SP600125 concentration of JNK pathway of skin fibroblasts activated by UVA, and to lay a foundation for studying the mechanism of JNK signaling pathway in photoaging. [methods] Primary cultured skin fibroblasts from prepuce of children were cultured. The effects of different doses of UVA, different concentrations of SP600125 and their combination on the activity and morphology of skin fibroblasts were studied by CCK-8 method and fluorescence inverted microscope, in order to select the non-cytotoxic UVA dose and SP600125 concentration. Then Western-blot was used to detect the phosphorylated c-Jun-P-c-Jun-Jun expression in skin fibroblasts at different time points after UVA irradiation to determine the time point at which it was induced to express. Finally, Western-blot was used to detect the inhibitory effect of different concentrations of SP600125 on the up-regulated P-c-Jun expression of UVA. [results] UVA and SP600125 irradiated cells and incubated cells with UVA 鈮，

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