中国病理生理杂志
中國病理生理雜誌
중국병리생리잡지
CHINESE JOURNAL OF PATHOPHYSIOLOGY
2009年
12期
2329-2333
,共5页
魏水生%廖新学%谭钰嫔%杨战利%杨春涛%赵春梅%董小变%王礼春%陈培熹%冯鉴强
魏水生%廖新學%譚鈺嬪%楊戰利%楊春濤%趙春梅%董小變%王禮春%陳培熹%馮鑒彊
위수생%료신학%담옥빈%양전리%양춘도%조춘매%동소변%왕례춘%진배희%풍감강
热休克蛋白90%硫化氢%氯化钴%心肌细胞%氧化性应激
熱休剋蛋白90%硫化氫%氯化鈷%心肌細胞%氧化性應激
열휴극단백90%류화경%록화고%심기세포%양화성응격
Heat-shock proteins 90%Hydrogen sulfide%Cobalt chloride%Cardiomyocytes%Oxidative stress
目的:探讨热休克蛋白90(HSP90)在硫化氢(H_2S)对抗化学性低氧模拟剂氯化钴(CoCl_2)诱导H9c2心肌细胞氧化应激损伤中的作用.方法: 应用CoCl_2处理H9c2心肌细胞,建立化学性缺氧损伤心肌细胞的实验模型.在CoCl_2处理H9c2心肌细胞前30 min,把硫氢化钠(NaHS,H_2S的供体)加入培养基中,作为预处理.应用高效液相色谱法(HPLC)检测细胞内ATP的含量;罗丹明123(Rh123)染色荧光显微镜照相检测线粒体膜电位(MMP);超氧化物歧化酶(SOD)检测试剂盒检测SOD活性;免疫印迹法(Western blotting)检测血红素氧合酶-1(HO-1)的表达.结果: 600 μmol/L CoCl_2明显地降低H9c2心肌细胞内SOD活性、ATP水平及MMP,并增加HO-1表达.400 μmol/L NaHS 预处理可显著地抑制CoCl_2诱导的细胞毒性及氧化应激反应,使SOD活性、ATP水平及MMP提高,HO-1表达减少.热休克蛋白90抑制剂17-丙烯胺基-17去甲氧基格尔德霉素(17AAG)能明显地阻断H_2S对CoCl_2诱导的细胞毒性和氧化应激反应的抑制作用,使细胞内ATP水平及MMP降低,HO-1表达增多,但对SOD活性的影响不明显.结论: 热休克蛋白90可通过抑制化学性缺氧引起的氧化应激反应来介导H_2S的心肌保护作用.
目的:探討熱休剋蛋白90(HSP90)在硫化氫(H_2S)對抗化學性低氧模擬劑氯化鈷(CoCl_2)誘導H9c2心肌細胞氧化應激損傷中的作用.方法: 應用CoCl_2處理H9c2心肌細胞,建立化學性缺氧損傷心肌細胞的實驗模型.在CoCl_2處理H9c2心肌細胞前30 min,把硫氫化鈉(NaHS,H_2S的供體)加入培養基中,作為預處理.應用高效液相色譜法(HPLC)檢測細胞內ATP的含量;囉丹明123(Rh123)染色熒光顯微鏡照相檢測線粒體膜電位(MMP);超氧化物歧化酶(SOD)檢測試劑盒檢測SOD活性;免疫印跡法(Western blotting)檢測血紅素氧閤酶-1(HO-1)的錶達.結果: 600 μmol/L CoCl_2明顯地降低H9c2心肌細胞內SOD活性、ATP水平及MMP,併增加HO-1錶達.400 μmol/L NaHS 預處理可顯著地抑製CoCl_2誘導的細胞毒性及氧化應激反應,使SOD活性、ATP水平及MMP提高,HO-1錶達減少.熱休剋蛋白90抑製劑17-丙烯胺基-17去甲氧基格爾德黴素(17AAG)能明顯地阻斷H_2S對CoCl_2誘導的細胞毒性和氧化應激反應的抑製作用,使細胞內ATP水平及MMP降低,HO-1錶達增多,但對SOD活性的影響不明顯.結論: 熱休剋蛋白90可通過抑製化學性缺氧引起的氧化應激反應來介導H_2S的心肌保護作用.
목적:탐토열휴극단백90(HSP90)재류화경(H_2S)대항화학성저양모의제록화고(CoCl_2)유도H9c2심기세포양화응격손상중적작용.방법: 응용CoCl_2처리H9c2심기세포,건립화학성결양손상심기세포적실험모형.재CoCl_2처리H9c2심기세포전30 min,파류경화납(NaHS,H_2S적공체)가입배양기중,작위예처리.응용고효액상색보법(HPLC)검측세포내ATP적함량;라단명123(Rh123)염색형광현미경조상검측선립체막전위(MMP);초양화물기화매(SOD)검측시제합검측SOD활성;면역인적법(Western blotting)검측혈홍소양합매-1(HO-1)적표체.결과: 600 μmol/L CoCl_2명현지강저H9c2심기세포내SOD활성、ATP수평급MMP,병증가HO-1표체.400 μmol/L NaHS 예처리가현저지억제CoCl_2유도적세포독성급양화응격반응,사SOD활성、ATP수평급MMP제고,HO-1표체감소.열휴극단백90억제제17-병희알기-17거갑양기격이덕매소(17AAG)능명현지조단H_2S대CoCl_2유도적세포독성화양화응격반응적억제작용,사세포내ATP수평급MMP강저,HO-1표체증다,단대SOD활성적영향불명현.결론: 열휴극단백90가통과억제화학성결양인기적양화응격반응래개도H_2S적심기보호작용.
AIM: To explore the roles of heat shock protein 90 (HSP90) in the blockage of hydrogen sulfide (H2S) against chemical hypoxia-mimetic agent (cobalt chloride, CoCl_2)-induced oxidative stress injuries in H9c2 cardiac cell. METHODS: H9c2 cells were treated with CoCl_2 to set up the chemical hypoxia-induced the model of cardiomyocyte injury. Sodium hydrosulfide (NaHS, a H2S donor) was added into medium for 30 min before CoCl_2 treatment. ATP content was detected by high performance liquid chromatogram (HPLC). Mitochondrial membrane potential (MMP) was measured by rhodamine123 (Rh123) staining and photofluorography. The activity of superoxide dismutase (SOD) was observed using a SOD kit. The expression of heme oxygenase-1 (HO-1) was evaluated by Western blotting. RESULTS: CoCl_2 at concentration of 600 μmol/L significantly reduced SOD activity, ATP level and MMP, and enhanced the expression of HO-1 in H9c2 cells. Pretreatment with 400 μmol/L NaHS dramatically inhibited the cytotoxicity induced by CoCl_2, increased SOD activity, ATP level and MMP, decreased HO-1 expression. 17-allylamino-17 demethoxygeldanamycine(17AAG), an inhibitor of HSP90, obviously blocked the inhibitory effect of H2S on the CoCl_2-induced cytotoxicity, reduced the levels of ATP and MMP, increased HO-1 expression. However, no significantly influence on SOD activity was observed. CONCLUSION: HSP90 may mediate the cardioprotection of H2S via inhibiting the oxidative stress induced by chemical hypoxia.
