Chen Lan, Li Hui, Li Chun, Cao Jia-hui, Wu Yi-ling, Liu Yang, Zhou Jian-hong, Chen Dong-feng, Li Yi-wei
Teaching and Research Section of Anatomy, Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China.
Zhong Xi Yi Jie He Xue Bao. 2011 Aug;9(8):888-93. doi: 10.3736/jcim20110811.
To investigate the effects of active ingredients of Plastrum Testudinis (PT) on serum deprivation-induced apoptosis of epidermal stem cells (ESCs).
ESCs were isolated from the back skin of fetal Sprague-Dawley rats with 2 weeks of gestational age and were divided into normal group (10% fetal bovine serum), control group (serum-deprived culture) and groups treated with serum deprivation plus active ingredients of PT, including ethyl acetate extract (2B), stearic acid ethyl ester (S6), tetradecanoic acid sterol ester (S8) and (+)-4-cholesten-3-one (S9). The vitality of ESCs after 24, 48 and 72 h of culture was measured with MTT method; apoptotic ESCs double-stained with Annexin V-FITC and propidium iodine were detected by flow cytometry (FCM); Bcl-2 and caspase-3 expressions were measured by Western blotting.
MTT results indicated that the vitality of ESCs in the active ingredients of PT groups at 48 h was increased compared with the control group and 2B had better effects than the others. FCM results indicated that 2B had the most significant anti-apoptotic effect compared with the control as well as S6, S8 and S9. Western blot results indicated that 2B, S6, S8 and S9 up-regulated the expression of Bcl-2 protein and down-regulated the expression of caspase-3 protein compared with the control.
Ethyl acetate extract of Plastrum Testudinis inhibits epidermal stem cell apoptosis in serum-deprived culture by regulating the expressions of Bcl-2 and caspase-3 proteins and has a stronger anti-apoptotic effect than its constituents S6, S8 and S9.
探讨龟板活性成分对血清剥夺诱导的表皮干细胞（ESCs）凋亡的影响。
从孕龄2周的Sprague-Dawley胎鼠背部皮肤分离ESCs，分为正常组（10%胎牛血清）、对照组（血清剥夺培养）以及血清剥夺加龟板活性成分处理组，活性成分包括乙酸乙酯提取物（2B）、硬脂酸乙酯（S6）、十四烷酸甾醇酯（S8）和（+）-4-胆甾烯-3-酮（S9）。采用MTT法检测培养24、48和72 h后ESCs的活力；通过流式细胞术（FCM）检测用Annexin V-FITC和碘化丙啶双染的凋亡ESCs；采用蛋白质印迹法检测Bcl-2和caspase-3的表达。
MTT结果显示，与对照组相比，48 h时龟板活性成分组ESCs的活力增加，且2B的效果优于其他组。FCM结果表明，与对照组以及S6、S8和S9相比，2B具有最显著的抗凋亡作用。蛋白质印迹结果表明，与对照组相比，2B、S6、S8和S9上调Bcl-2蛋白的表达，下调caspase-3蛋白的表达。
龟板乙酸乙酯提取物通过调节Bcl-2和caspase-3蛋白的表达抑制血清剥夺培养中的表皮干细胞凋亡，且其抗凋亡作用强于其成分S6、S8和S9。
