白藜芦醇以Caspase依赖和非依赖两种方式导致Jurkat细胞死亡

应用形态学观察、流式细胞仪检测、Western印迹和DNA凝胶电泳等方法研究白藜芦醇对Jurkat细胞的作用。发现白藜芦醇处理组中细胞有皱缩、出泡、染色质边集等现象,但染色质浓缩呈散在团块状且不致密。细胞质结构疏松,线粒体肿胀,脊消失。少见凋亡小体。在白藜芦醇处理组,Western印迹可检测到弱的17kDacaspase-3条带,DNA凝胶电泳可以检测到梯状DNA和弥散条带;流式细胞仪在白藜芦醇处理组检测到大量PI单阳性细胞和少量膜联蛋白V单阳性细胞。Z-VAD-FMK干预后可以发现细胞死亡率降低,同时该组梯状DNA消失,但是大分子量弥散DNA条带依然可以检测到。结果表明白藜芦醇可以通过caspase依赖和非依赖途径导致Jurkat细胞死亡。此分子机制的明确将为白藜芦醇应用于临床白血病的治疗打下理论基础。

Resveratrol Triggers both Caspase Independent and Dependent Cell Death Pathway in T Lymphoblast Leukemia Jurkat Cells

To study effects of resveratrol on acute lymphoblast leukemia Jurkat cell line,morphologic changes were observed under light microscope,fluorescence microscope and electron microscope.Cell death was determined by phosphatidylserine translocation and mitochondrial membrane permeabilization with flow cytometry(FCM).Western blot was conducted to detect the activation of caspase-3.DNA ladder was detected with gel electrophoresis.In resveratrol treatment groups,cell shrinkage,blebbing,margination and lumpy chromatin condensation,mitochondrial swelling and cristae membrane loss were observed under light microscope and electron microscope in treatment groups.More PI positive cells versus fewer annexin V positive cells were detected in 0.2 mmol/L and higher concentration groups at 24 h and 36 h check points in FCM evaluation.Western blot detected the activated 17 kDa caspase-3 in all treatment groups while negative in control.Cell death can be partly inhibited by pan-caspase inhibitor,Z-VAD-FMK.Gel electrophoresis displayed both DNA ladder and smear in resveratrol ex-posed groups,while DNA ladder disappeared in Z-VAD-FMK inhibiting group and left only smear.The above data suggest that resveratrol can induce both caspase dependent and independent cell death of Jurkat cells.Molecular understanding of resveratrol in Jurkat cell will be helpful in its future clinic application.