中国医药生物技术
中國醫藥生物技術
중국의약생물기술
CHINESE MEDICINAL BIOTECHNOLOGY
2014年
6期
438-445
,共8页
内皮细胞%氧化性应激%儿茶酚类%吉西他滨
內皮細胞%氧化性應激%兒茶酚類%吉西他濱
내피세포%양화성응격%인다분류%길서타빈
Endothelial cells%Oxidative stress%Catechols%Gemcitabine
目的观察吉西他滨对大鼠肺微血管内皮细胞的影响及表没食子儿茶素没食子酸酯的保护作用。<br> 方法利用植块法分离大鼠肺微血管内皮细胞。SRB 法检测药物对肺微血管内皮细胞和 A549增殖的影响。流式细胞仪用于检测细胞周期和细胞凋亡。Western blot 法检测凋亡相关蛋白含量。测定细胞内活性氧和超氧化物歧化酶水平以及乳酸脱氢酶渗透情况。<br> 结果吉西他滨能显著减少肺微血管内皮细胞细胞增殖并将其阻滞于 S 期。吉西他滨引起肺微血管内皮细胞细胞凋亡具有时间依赖性，处理24、48、72 h 后细胞凋亡率分别为7.2%、15.4%、23.3%。同时，吉西他滨作用后细胞内活性氧含量上升，而处理48 h 后超氧化物歧化酶水平显著下降。进一步研究发现，表没食子儿茶素没食子酸酯能够提高吉西他滨处理后肺微血管内皮细胞的存活率，并且减弱吉西他滨造成的超氧化物歧化酶水平的下降。<br> 结论吉西他滨对肺微血管内皮细胞具有损伤作用，能够引起肺微血管内皮细胞的凋亡和氧化应激的发生，而表没食子儿茶素没食子酸酯对吉西他滨引起的肺微血管内皮细胞损伤具有保护作用。
目的觀察吉西他濱對大鼠肺微血管內皮細胞的影響及錶沒食子兒茶素沒食子痠酯的保護作用。<br> 方法利用植塊法分離大鼠肺微血管內皮細胞。SRB 法檢測藥物對肺微血管內皮細胞和 A549增殖的影響。流式細胞儀用于檢測細胞週期和細胞凋亡。Western blot 法檢測凋亡相關蛋白含量。測定細胞內活性氧和超氧化物歧化酶水平以及乳痠脫氫酶滲透情況。<br> 結果吉西他濱能顯著減少肺微血管內皮細胞細胞增殖併將其阻滯于 S 期。吉西他濱引起肺微血管內皮細胞細胞凋亡具有時間依賴性，處理24、48、72 h 後細胞凋亡率分彆為7.2%、15.4%、23.3%。同時，吉西他濱作用後細胞內活性氧含量上升，而處理48 h 後超氧化物歧化酶水平顯著下降。進一步研究髮現，錶沒食子兒茶素沒食子痠酯能夠提高吉西他濱處理後肺微血管內皮細胞的存活率，併且減弱吉西他濱造成的超氧化物歧化酶水平的下降。<br> 結論吉西他濱對肺微血管內皮細胞具有損傷作用，能夠引起肺微血管內皮細胞的凋亡和氧化應激的髮生，而錶沒食子兒茶素沒食子痠酯對吉西他濱引起的肺微血管內皮細胞損傷具有保護作用。
목적관찰길서타빈대대서폐미혈관내피세포적영향급표몰식자인다소몰식자산지적보호작용。<br> 방법이용식괴법분리대서폐미혈관내피세포。SRB 법검측약물대폐미혈관내피세포화 A549증식적영향。류식세포의용우검측세포주기화세포조망。Western blot 법검측조망상관단백함량。측정세포내활성양화초양화물기화매수평이급유산탈경매삼투정황。<br> 결과길서타빈능현저감소폐미혈관내피세포세포증식병장기조체우 S 기。길서타빈인기폐미혈관내피세포세포조망구유시간의뢰성，처리24、48、72 h 후세포조망솔분별위7.2%、15.4%、23.3%。동시，길서타빈작용후세포내활성양함량상승，이처리48 h 후초양화물기화매수평현저하강。진일보연구발현，표몰식자인다소몰식자산지능구제고길서타빈처리후폐미혈관내피세포적존활솔，병차감약길서타빈조성적초양화물기화매수평적하강。<br> 결론길서타빈대폐미혈관내피세포구유손상작용，능구인기폐미혈관내피세포적조망화양화응격적발생，이표몰식자인다소몰식자산지대길서타빈인기적폐미혈관내피세포손상구유보호작용。
Objective To investigate the damage of gemcitabine on rat pulmonary microvascular endothelial cells and the protective effect of EGCG. <br> Methods PMVECs were isolated by modification of a tissue explant method. SRB assay was used to investigate the growth of PMVECs and A549 cells. Flow cytometry was applied to measure the cell cycle and apoptosis. Western blot was used to determine the content of apoptosis-related proteins. The activities of ROS, SOD and LDH were examined with microplate reader. <br> Results The exposure to gemcitabine decreased the survival of PMVECs and arrested PMVECs at S phase. Gemcitabine increased the number of apoptotic cells time-dependently and the percentage of apoptotic cells treated for 24, 48, 72 h were 7.2%, 15.4%, 23.3%, respectively. Meanwhile, intracellular ROS level of PMVECs was elevated and after 48 h the SOD level was lowered markedly after gemcitabine treatment. Further investigation revealed that EGCG increased cell viability of PMVECs with or without gemcitabine and augmented the intracellular SOD level of PMVECs reduced by gemcitabine. <br> Conclusion These results suggest that gemcitabine injures PMVECs by inducing pulmonary endothelial cell apoptosis and oxidative stress. EGCG exerts the protective effect on gemcitabine-induced endothelial cells injury.
