华西医科大学学报
華西醫科大學學報
화서의과대학학보
JOURNAL OF WEST CHINA UNIVERSITY OF MEDICAL SCIENCES
2001年
2期
208-212
,共5页
彭黎明%张芹%江虹%Christopher J Bradely%James S Wiley
彭黎明%張芹%江虹%Christopher J Bradely%James S Wiley
팽려명%장근%강홍%Christopher J Bradely%James S Wiley
受体%嘌呤能P2Z%细胞凋亡%慢性淋巴细胞白血病%二价阳离子
受體%嘌呤能P2Z%細胞凋亡%慢性淋巴細胞白血病%二價暘離子
수체%표령능P2Z%세포조망%만성림파세포백혈병%이개양리자
目的探讨嘌呤能P2Z受体介导慢性淋巴细胞白血病（CLL）细胞凋亡的影响因素及其机理。方法在二价阳离子——1.0 mmol/L Mg2+、Zn2+、Ca2+、Sr2+、Co2+、Ba2+,不同浓度的EDTA 或EGTA，不同温度及在含150 mmol/L胆碱的介质中，将表达P2Z受体［P2Z（+）］的CLL 细胞分别同1.0 mmol/L 三磷酸腺苷（ATP）或0.1 mmol/L 苯甲酰苯甲酸ATP（BzATP）体外培养8小时，以DNA凝胶电泳、TdT法和流式细胞分析（FCA）检测上述条件下细胞凋亡的诱导或抑制效应。结果 Mg2+或Ca2+能以剂量依赖性方式促进ATP诱导P2Z（+）细胞凋亡，而EDTA 或EGTA却以相反的方式抑制P2Z（+）细胞凋亡的发生；1.0 mmol/L Zn2+可完全阻止ATP诱导P2Z（+）细胞凋亡所产生的DNA片段，但其它二价阳离子包括1.0 mmol/L Sr2+、Co2+、Ba2+却不影响ATP的诱导；胆碱作为磷脂酶D（PLD）的抑制剂，也可部分抑制P2Z（+）细胞凋亡产生的DNA片段；当温度低于10℃，可完全阻止ATP诱导P2Z（+）细胞凋亡产生DNA片段的发生。结论 P2Z受体介导CLL细胞凋亡可能与核酸内切酶，PLD的参与密切相关。
目的探討嘌呤能P2Z受體介導慢性淋巴細胞白血病（CLL）細胞凋亡的影響因素及其機理。方法在二價暘離子——1.0 mmol/L Mg2+、Zn2+、Ca2+、Sr2+、Co2+、Ba2+,不同濃度的EDTA 或EGTA，不同溫度及在含150 mmol/L膽堿的介質中，將錶達P2Z受體［P2Z（+）］的CLL 細胞分彆同1.0 mmol/L 三燐痠腺苷（ATP）或0.1 mmol/L 苯甲酰苯甲痠ATP（BzATP）體外培養8小時，以DNA凝膠電泳、TdT法和流式細胞分析（FCA）檢測上述條件下細胞凋亡的誘導或抑製效應。結果 Mg2+或Ca2+能以劑量依賴性方式促進ATP誘導P2Z（+）細胞凋亡，而EDTA 或EGTA卻以相反的方式抑製P2Z（+）細胞凋亡的髮生；1.0 mmol/L Zn2+可完全阻止ATP誘導P2Z（+）細胞凋亡所產生的DNA片段，但其它二價暘離子包括1.0 mmol/L Sr2+、Co2+、Ba2+卻不影響ATP的誘導；膽堿作為燐脂酶D（PLD）的抑製劑，也可部分抑製P2Z（+）細胞凋亡產生的DNA片段；噹溫度低于10℃，可完全阻止ATP誘導P2Z（+）細胞凋亡產生DNA片段的髮生。結論 P2Z受體介導CLL細胞凋亡可能與覈痠內切酶，PLD的參與密切相關。
목적탐토표령능P2Z수체개도만성림파세포백혈병（CLL）세포조망적영향인소급기궤리。방법재이개양리자——1.0 mmol/L Mg2+、Zn2+、Ca2+、Sr2+、Co2+、Ba2+,불동농도적EDTA 혹EGTA，불동온도급재함150 mmol/L담감적개질중，장표체P2Z수체［P2Z（+）］적CLL 세포분별동1.0 mmol/L 삼린산선감（ATP）혹0.1 mmol/L 분갑선분갑산ATP（BzATP）체외배양8소시，이DNA응효전영、TdT법화류식세포분석（FCA）검측상술조건하세포조망적유도혹억제효응。결과 Mg2+혹Ca2+능이제량의뢰성방식촉진ATP유도P2Z（+）세포조망，이EDTA 혹EGTA각이상반적방식억제P2Z（+）세포조망적발생；1.0 mmol/L Zn2+가완전조지ATP유도P2Z（+）세포조망소산생적DNA편단，단기타이개양리자포괄1.0 mmol/L Sr2+、Co2+、Ba2+각불영향ATP적유도；담감작위린지매D（PLD）적억제제，야가부분억제P2Z（+）세포조망산생적DNA편단；당온도저우10℃，가완전조지ATP유도P2Z（+）세포조망산생DNA편단적발생。결론 P2Z수체개도CLL세포조망가능여핵산내절매，PLD적삼여밀절상관。
Objective To explore the effects of various factors affecting apoptosis of chronic lymphocytic leukemia (CLL) cells mediated by P2Z purinoceptor. Methods In vitro exposure of CLL cells with P2Z receptors to 1 mmol/L ATP or 0.1 mmol/L benzoylbenzoic ATP (BzATP) for 8 h in the presence of 1.0 mmol/L Mg2+ or other bivalent cations, 150 mmol/L choline in media, 1,2,4 mmol/L EDTA or EGTA, or under different temperature. ATP-induced apoptosis was detected by agarose gel electrophoresis, TdT assay and flow cytometric analysis (FCA). Results The results showed that ① Extracellular Mg2+ or Ca2+ stimulated ATP-induced DNA fragmentation in a dose-dependent manner, and the compatible evidence was provided by the inhibition of ATP-induced DNA fragmentation in the presence of EGTA or EDTA; ② ATP-induced DNA fragmentation was completely inhibited by 1.0 mmol/L Zn2+; ③ ATP-induced DNA breaks were not affected by Ba2+, Sr2+, Co2+ when they were substituted for extracellular Mg2+ or Ca2+; ④ Choline, an inhibitor of phospholipase D (PLD) stimulated by ATP through P2Z receptor in human lymphocytes, was also a partial inhibitor of ATP-induced DNA fragmentation, and the results were confirmed by flow cytometric analysis (FCA); ⑤ ATP-induced DNA fragmentation was completely obliterated when the temperature was lower than 10℃. Conclusion These results suggest that the endonuclease and PLD may be involved in ATP-induced apoptosis in human lymphocytes via P2Z receptor.
