中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2011年
5期
727-729
,共3页
胶质瘤%声动力治疗%血卟啉甲醚%脱噬作用
膠質瘤%聲動力治療%血卟啉甲醚%脫噬作用
효질류%성동력치료%혈계람갑미%탈서작용
Glioma%Sonodynamic therapy%Hematoporphyrin monomethy lether%Apoptosis
目的 探讨血卟啉甲醚声动力疗法(SDT)对体外C6胶质瘤细胞的促凋亡作用及机制.方法 取处于对数生长期的C6胶质瘤细胞接种于96孔、6孔培养板上.采用噻唑蓝(MTT)比色法观察在不同时间超声波辐照后细胞存活率;流式细胞仪检测24 h后对照组、血卟啉甲醚组(HMME)、单纯超声组、超声血卟啉组(SDT)C6胶质瘤细胞凋亡率、活性氧(ROS)产生、线粒体膜电位(MMP)的改变;Western blot检测凋亡蛋白bcl-2、bax、Caspase-3、-9的表达及细胞色素C(Cyt-C)的释放.结果 当超声频率为1.0 mHz,声强为1.0 W/cm2,MTT筛选60 s为最佳辐照时间.与对照组、HMME及单纯超声组比较,SDT组凋亡率(34.7±1.9)%明显增高(P＜0.05),同时伴有ROS(36.4±3.1)%的产生和MMP(44.9±3.4)%的降低.SDT组Cyt-C的释放增加;凋亡蛋白bax、Caspase-3、-9表达水平增高,凋亡蛋白bcl-2表达水平减低.结论 血卟啉甲醚声动力诱导C6胶质瘤细胞凋亡过程中伴有ROS的产生和MMP的减低,线粒体途径对C6胶质瘤细胞凋亡起着重要作用.
目的 探討血卟啉甲醚聲動力療法(SDT)對體外C6膠質瘤細胞的促凋亡作用及機製.方法 取處于對數生長期的C6膠質瘤細胞接種于96孔、6孔培養闆上.採用噻唑藍(MTT)比色法觀察在不同時間超聲波輻照後細胞存活率;流式細胞儀檢測24 h後對照組、血卟啉甲醚組(HMME)、單純超聲組、超聲血卟啉組(SDT)C6膠質瘤細胞凋亡率、活性氧(ROS)產生、線粒體膜電位(MMP)的改變;Western blot檢測凋亡蛋白bcl-2、bax、Caspase-3、-9的錶達及細胞色素C(Cyt-C)的釋放.結果 噹超聲頻率為1.0 mHz,聲彊為1.0 W/cm2,MTT篩選60 s為最佳輻照時間.與對照組、HMME及單純超聲組比較,SDT組凋亡率(34.7±1.9)%明顯增高(P＜0.05),同時伴有ROS(36.4±3.1)%的產生和MMP(44.9±3.4)%的降低.SDT組Cyt-C的釋放增加;凋亡蛋白bax、Caspase-3、-9錶達水平增高,凋亡蛋白bcl-2錶達水平減低.結論 血卟啉甲醚聲動力誘導C6膠質瘤細胞凋亡過程中伴有ROS的產生和MMP的減低,線粒體途徑對C6膠質瘤細胞凋亡起著重要作用.
목적 탐토혈계람갑미성동력요법(SDT)대체외C6효질류세포적촉조망작용급궤제.방법 취처우대수생장기적C6효질류세포접충우96공、6공배양판상.채용새서람(MTT)비색법관찰재불동시간초성파복조후세포존활솔;류식세포의검측24 h후대조조、혈계람갑미조(HMME)、단순초성조、초성혈계람조(SDT)C6효질류세포조망솔、활성양(ROS)산생、선립체막전위(MMP)적개변;Western blot검측조망단백bcl-2、bax、Caspase-3、-9적표체급세포색소C(Cyt-C)적석방.결과 당초성빈솔위1.0 mHz,성강위1.0 W/cm2,MTT사선60 s위최가복조시간.여대조조、HMME급단순초성조비교,SDT조조망솔(34.7±1.9)%명현증고(P＜0.05),동시반유ROS(36.4±3.1)%적산생화MMP(44.9±3.4)%적강저.SDT조Cyt-C적석방증가;조망단백bax、Caspase-3、-9표체수평증고,조망단백bcl-2표체수평감저.결론 혈계람갑미성동력유도C6효질류세포조망과정중반유ROS적산생화MMP적감저,선립체도경대C6효질류세포조망기착중요작용.
Objective To study apoptotic effect and mechanisms of sonodynamic therapy (SDT) with hematoporphyrin monomethyl ether (HMME) on C6 glioma cells in vitro. Methods Time of ultrasound irradiation was optimized by methyl thiazol tetrazolium (MTT) . Apoptotic rate( AR) generation of reactive oxygen species (ROS) ,mitochondrial membrane potential (MMP) were detected by flow cytometry ,and expressions of apoptosis proteins were measured by Western blotting after SDT treatment. Results 60 s is the optimal time irradiation by MTT when the ultrasound frequency was maintained in 1 mHz and the average intensity was kept in 1. 0 W/cm2. To compare with the control,HMME and ultrasound group,treatment of C6 glioma cells with SDT resulted in the occurrence of apoptosis(34. 7 ±1.9)% .which was associated with the production of ROS ( 36. 4 ± 3. 1) % and loss of MMP (44. 9 ± 3. 4) %. The results showed that significantly increased expression in bax,Caspase-9 ,-3 , decreased in bcl-2 after SDT treatment. The results also showed apparent release of cytochome c in the SDT group versus to the other three groups. Conclusion Our results suggested that SDT mediated by HMME could induce apoptosis in C6 glioma cells.The mitochondrial signal pathway may play a pivotal role in the apoptosis.
