CAJ | 학술논문

[目的]探讨硫化氢(H_2S)对抗二氯化钴(CoCl_2)诱导PC12细胞凋亡的作用及其初步机制.[方法]应用化学性缺氧模拟剂CoCl_2在PC12细胞建立化学性缺氧损伤模型;以硫氢化钠(NaHS)作为H_2S的供体;应用CCK-8比色法检测细胞存活率;碘化丙啶(PI)染色流式细胞技术(FCM)检测细胞凋亡率;Hoechst33258染色检测细胞凋亡的形态学变化;罗丹明123(Rh123)染色荧光显微镜照相检测细胞线粒体膜电位(MMP);2',7'-二氯荧光黄双乙酸盐(DCFH-DA)染色荧光显微镜照相检测细胞内的活性氧(ROS)水平.[结果] COCl_2引起PC12细胞的存活率显著降低及凋亡率明显增加,同时引起PC12细胞的MMP明显降低及ROS生成显著增加.在600μmol/L CoCl_2处理PC12细胞前,应用100～400 μmol/L NaHS预处理30 min,呈浓度依赖性地阻断CoCl_2引起PC12细胞的存活率降低;200 μmol/L、400 μmol/L NaHS预处理能显著地降低600 μmol/L CoCl_2引起PC12细胞的凋亡率增加并阻断CoCl_2引起的MMP降低及ROS升高.[结论]H_2S具有神经细胞保护作用,能明显地对抗CoCl_2诱导的PC12细胞凋亡,此作用可能与其减少ROS生成,提高MMP有关.
[목적]탐토류화경(H_2S)대항이록화고(CoCl_2)유도PC12세포조망적작용급기초보궤제.[방법]응용화학성결양모의제CoCl_2재PC12세포건립화학성결양손상모형;이류경화납(NaHS)작위H_2S적공체;응용CCK-8비색법검측세포존활솔;전화병정(PI)염색류식세포기술(FCM)검측세포조망솔;Hoechst33258염색검측세포조망적형태학변화;라단명123(Rh123)염색형광현미경조상검측세포선립체막전위(MMP);2',7'-이록형광황쌍을산염(DCFH-DA)염색형광현미경조상검측세포내적활성양(ROS)수평.[결과] COCl_2인기PC12세포적존활솔현저강저급조망솔명현증가,동시인기PC12세포적MMP명현강저급ROS생성현저증가.재600μmol/L CoCl_2처리PC12세포전,응용100～400 μmol/L NaHS예처리30 min,정농도의뢰성지조단CoCl_2인기PC12세포적존활솔강저;200 μmol/L、400 μmol/L NaHS예처리능현저지강저600 μmol/L CoCl_2인기PC12세포적조망솔증가병조단CoCl_2인기적MMP강저급ROS승고.[결론]H_2S구유신경세포보호작용,능명현지대항CoCl_2유도적PC12세포조망,차작용가능여기감소ROS생성,제고MMP유관.
[Objective] To explore the cytoprotecfion of hydrogen sulfide (H_2S) against cobalt chloride (CoCl_2)-induced apeptosis in PC12 cells and the underlying mechanisms. [Methods] CoCl_2 (a chemical hypoxia-mimetic agent) was used to establish the chemical hypoxia-induced PC12 cell injuries model. Sodium hydrosulfide (NaHS) was used as a H_2S donor. The viability of PC12 cells was measured by CCK-8 assay. The percentage of apeptotic cells was assessed by propidium iodide stain flow cytometry (FCM). The morphological change of apeptotic cells was tested by using the chromatin dye Hoechst 33258. The mitochondrial membrane potential (MMP) was analyzed by rhodamine 123 staining and photofluorography. The level of reactive oxygen species (ROS) in PC12 cells was measured by DCFH-DA staining and photofluorography. [Results] CoCl_2 induced a decrease in cell viability and an increase in percentage of apeptosis in PC12 cells along with dissipation of MMP as well as overproduction of ROS. When PC12 cells were treated with Naris 30 min before CoCl_2 treatment a decrease in viability of PC12 cells induced by 600 μmol/L CoCl_2 was concentration-dependently blocked by NaHs (100, 200, and 400 μmol/L). Pretreatment with NaHS at 200 and 400 μmol/L obviously reduced the apepetotic percentage of PC12 cells induced by 600 μmol/L CoCl_2 and inhibited the dissipation of MMP and overproduction of ROS. [Conclusion] H_2S protected PC12 cells against CoCl_2-induced apeptosis, which may be associated with the inhibition of H_2S on the dissipation of MMP and overproduction of ROS induced by CoCl_2.