Known as Ahle ons DNA fragmentation in Maraviroc Selzentry apoptotic cell death after the cleavage between nucleosomal activated endonuclease. The cell morphology was visualized by a DNA-binding dye Hoechst 33258 membranepermeable. PC12 cells without exposure to drugs were uniformly Ig found with the exception of a few abnormal nuclei Rbt. after treatment with H 12 Z ligustilide and dopamine, alone or in combination, the morphology of the nucleus of claim Hoechst 33258-F is coloration clearly GE changed. Condensed chromatin was treated in about 45% of the cells with Z-ligustilide and dopamine in combination w While only 10.3 and 21.7% cells in each treated with dopamine or Z ligustilide only. We further characterized the cell death caused by medication twice reqs Dyeing the cells with annexin V and PI induced. After 18 h of treatment with Z ligustilide and dopamine, alone or in combination, PC12 cells were analyzed by flow cytometry and classified early stage of apoptosis sp T apoptotic cells / necrotic and lebensf HIGEN cells. As shown in Fig. 5, the sum of the first apoptotic cells and apoptotic end / necrotic cells as high as 56.8% of the cells with Z-ligustilide and dopamine was treated in combination, w While only 19.5 and 31.3% of the cells each with Z ligustilide dopamine alone or treated. The cytotoxicity t of Z ligustilide and dopamine by synergistic erh Increase the intracellular Ren ROS formation and degradation of GSH to the question of how the cytotoxicity t of Z ligustilide and dopamine answer will give the group, we examined the formation of intracellular Ren ROS and glutathione levels in cells treated with these drugs, singly and in combination. As shown in Fig. 6a, Z-ligustilide or dopamine alone decreased intracellular Re GSH levels to 55.6 and 63.2%, respectively, compared to untreated control cells.
Remarkably, the combination of Z ligustilide and Marbofloxacin dopamine significantly reduced the rate of intracellular Ren GSH to 28.8%. Such a decrease in the GSH level was treated as very important in the ratio Ratio to the content of GSH in the cells that produce dopamine were alone. Second, we have a cell permeable fluorescent probe DCFH DA to the level of intracellular Re ROS in response to Z ligustilide, dopamine, and the combination of the two drugs. Was induced on the basis of fluorescence intensity t of ROS were the intracellular Higher concentrations of ROS by 2.0 and 2.1 times increased Ht Z ligustilide and dopamine for 6 h betr Gt Remarkably, increases ht the combination of Z ligustilide and the intracellular dopamine Re ROS levels by 3.1 times compared with untreated control cells. The increase in intracellular Ren ROS levels by the combination was significantly compared to the caused by dopamine alone. Thiol-containing antioxidants prevent the synergistic cytotoxicity t of Z ligustilide and dopamine, we investigated the effect of thiol antioxidants N-acetylcysteine and GSH and not S Acid thiol antioxidants ascorbic Acid and trolox on the cytotoxicity t the combination Z ligustilide dopamine. According to the A.F. Apoptosis in the PC12 cells treated. In contrast, dopamine and Z ligustilide not induced synergistic cytotoxicity t in non-dopaminergic cells, such as human hepatocellular Ren carcinoma, HepG2 human epithelial carcinoma HeLa cells, human breast adenocarcinoma MCF-7 cells and human.