中国药理学通报
中國藥理學通報
중국약이학통보
Chinese Pharmacological Bulletin
2015年
10期
1421-1425,1426
,共6页
补体%补体旁路激活%内皮细胞%黏附分子%白藜芦醇%炎症反应%眼镜蛇毒因子
補體%補體徬路激活%內皮細胞%黏附分子%白藜蘆醇%炎癥反應%眼鏡蛇毒因子
보체%보체방로격활%내피세포%점부분자%백려호순%염증반응%안경사독인자
complement%complement alternative path-way ( activation )%endothelial cell%adhesion molecule%resveratrol%inflammation%cobra venom factor
目的：研究白藜芦醇( Res)、PDTC和AG4903种化学小分子对补体旁路激活致人微血管内皮细胞炎症反应相关黏附分子表达的干预作用和可能的作用机制。方法采用补体旁路激活产物作用于人微血管内皮细胞( HMEC )。TBA法检测细胞培养上清中丙二醛(MDA)的浓度,ELISA方法检测细胞培养上清中可溶性黏附分子ICAM-1、VCAM-1、E-selectin的表达,并采用多个浓度的Res、NF-κB信号通路抑制剂PDTC和JAK2信号通路抑制剂AG490预处理内皮细胞,研究3种抑制剂对细胞氧化水平和黏附分子表达的干预作用。利用Western blot检测补体旁路激活产物作用于内皮细胞对NF-κB p65磷酸化的影响及3种抑制剂的干预作用。结果 HMEC受补体旁路激活产物刺激后,导致细胞培养上清中MDA浓度升高, NF-κB p65磷酸化和黏附分子ICAM-1、VCAM-1和E-selectin的表达上调。 Res可降低细胞培养上清中MDA的浓度。 Res、PDTC和AG490能抑制NF-κB p65的磷酸化。 Res与PDTC能抑制内皮细胞黏附分子ICAM-1和VCAM-1的表达,对E-selectin的表达有一定的抑制作用,而AG490能够明显抑制上述3种黏附分子的表达。结论 Res、PDTC和AG490对补体旁路激活产物致内皮细胞黏附分子表达上调有不同程度的抑制作用,其作用机制与NF-κB信号通路的活化有关,而JAK2可能是一个更重要的调控位点。
目的：研究白藜蘆醇( Res)、PDTC和AG4903種化學小分子對補體徬路激活緻人微血管內皮細胞炎癥反應相關黏附分子錶達的榦預作用和可能的作用機製。方法採用補體徬路激活產物作用于人微血管內皮細胞( HMEC )。TBA法檢測細胞培養上清中丙二醛(MDA)的濃度,ELISA方法檢測細胞培養上清中可溶性黏附分子ICAM-1、VCAM-1、E-selectin的錶達,併採用多箇濃度的Res、NF-κB信號通路抑製劑PDTC和JAK2信號通路抑製劑AG490預處理內皮細胞,研究3種抑製劑對細胞氧化水平和黏附分子錶達的榦預作用。利用Western blot檢測補體徬路激活產物作用于內皮細胞對NF-κB p65燐痠化的影響及3種抑製劑的榦預作用。結果 HMEC受補體徬路激活產物刺激後,導緻細胞培養上清中MDA濃度升高, NF-κB p65燐痠化和黏附分子ICAM-1、VCAM-1和E-selectin的錶達上調。 Res可降低細胞培養上清中MDA的濃度。 Res、PDTC和AG490能抑製NF-κB p65的燐痠化。 Res與PDTC能抑製內皮細胞黏附分子ICAM-1和VCAM-1的錶達,對E-selectin的錶達有一定的抑製作用,而AG490能夠明顯抑製上述3種黏附分子的錶達。結論 Res、PDTC和AG490對補體徬路激活產物緻內皮細胞黏附分子錶達上調有不同程度的抑製作用,其作用機製與NF-κB信號通路的活化有關,而JAK2可能是一箇更重要的調控位點。
목적：연구백려호순( Res)、PDTC화AG4903충화학소분자대보체방로격활치인미혈관내피세포염증반응상관점부분자표체적간예작용화가능적작용궤제。방법채용보체방로격활산물작용우인미혈관내피세포( HMEC )。TBA법검측세포배양상청중병이철(MDA)적농도,ELISA방법검측세포배양상청중가용성점부분자ICAM-1、VCAM-1、E-selectin적표체,병채용다개농도적Res、NF-κB신호통로억제제PDTC화JAK2신호통로억제제AG490예처리내피세포,연구3충억제제대세포양화수평화점부분자표체적간예작용。이용Western blot검측보체방로격활산물작용우내피세포대NF-κB p65린산화적영향급3충억제제적간예작용。결과 HMEC수보체방로격활산물자격후,도치세포배양상청중MDA농도승고, NF-κB p65린산화화점부분자ICAM-1、VCAM-1화E-selectin적표체상조。 Res가강저세포배양상청중MDA적농도。 Res、PDTC화AG490능억제NF-κB p65적린산화。 Res여PDTC능억제내피세포점부분자ICAM-1화VCAM-1적표체,대E-selectin적표체유일정적억제작용,이AG490능구명현억제상술3충점부분자적표체。결론 Res、PDTC화AG490대보체방로격활산물치내피세포점부분자표체상조유불동정도적억제작용,기작용궤제여NF-κB신호통로적활화유관,이JAK2가능시일개경중요적조공위점。
Aim To investigate the effect of resveratrol ( Res) , PDTC and AG490 on adhesion molecules ex-pression induced by product of activated complement alternative pathway on human microvascular endothelial cells ( HMECs) and the possible mechanisms. Meth-ods HMECs were exposed to the product of comple-ment alternative pathway activation, then the superna-tant was removed to detect the concentration of malond-ialdehyde ( MDA ) with TBA method. ELISA method was used to detect the expression of soluble ICAM-1 , VCAM-1 ( and E-selectin) in the culture supernatant. Res, PDTC and AG490 with different concentrations were used to determine their effect on cell oxidation level and adhesion molecules expression. The phospho-rylation of NF-κB p65 was detected by Western blot, and the intervention of Res, PDTC and AG490 was as-sayed by the same way. Results The activation of complements alternative pathway resulted in the phos-phorylation of NF-κB p65 , and increased the concen-tration of MDA and up-regulated the expression of ICAM-1, VCAM-1 and E-selectin. Res reduced the concentration of MDA. Res, PDTC and AG490 inhibi-ted the phosphorylation of NF-κB p65 . Res and PDTC showed similar inhibition on expression of ICAM-1 and VCAM-1 , while exhibiting little effect on expression of E-selectin, and AG490 significantly inhibited the ex-pression of the above adhesion molecules. Conclusions Res, PDTC and AG490 could inhibit the expression of adhesion molecules induced by activated complement alternative pathway, the inhibition of NF-κB pathway activation was involved in their mechanism, and JAK2 may be a more important intervention target in regula-ting adhesion molecule expression.
