摘要：
目的:探討在模擬低氧環(huán)境下人白血病細胞株K562細胞增殖及低氧誘導(dǎo)因子(HIF-1α)表達的變化.方法:將白血病細胞株分為低氧處理組和常規(guī)對照組.對常規(guī)對照組細胞株進行常氧培養(yǎng);對低氧處理組用二氯化鈷(CoCl2) 50、200、400、800 μmol/L進行處理;在24、48和72 h后收集兩組細胞,并在倒置相差顯微下觀察細胞的形態(tài)變化,應(yīng)用MTT比色法檢測細胞增殖抑制率,實時熒光定量PCR檢測HIF-1α在轉(zhuǎn)錄水平的表達.結(jié)果:K562細胞形態(tài)在模擬低氧環(huán)境下隨CoCl2的濃度的增高和處理時間的延長而發(fā)生變形甚至細胞破裂等變化.MTT實驗結(jié)果顯示,細胞增殖在低氧環(huán)境下受到抑制,并且抑制作用隨CoC12的濃度增加和作用時間的延長而增強(r濃度=0.8712,r時間=0.6785).熒光定量PCR檢測結(jié)果顯示,細胞株的HIF-1αmRNA表達水平在CoCl2模擬低氧環(huán)境中均有上調(diào),并且對低氧時間和濃度呈依賴性,與CoCl2的處理濃度及處理時間呈正相關(guān)(r=0.954,r=0.895).結(jié)論:低氧環(huán)境可抑制白血病K562細胞的增殖并且使細胞中的低氧誘導(dǎo)因子HIF-1α的表達上調(diào),但未導(dǎo)致細胞發(fā)生誘導(dǎo)分化.%Objective:To explore the effect of hypoxia on the cell proliferation and expression of hypoxia-inducible factor-1α (HIF-1α) of human leukemia K562 cells.Methods:The leukemia cells were divided into 2 groups:hypoxiatreated group and conventional oxygen group as control.The K562 cells in hypoxia-treated group were treated with 50,200,400 and 800 μmol/L of CoCl2,while the K562 cells in control group were cultured in conventional oxygen condition,then the K562 cells in 2 groups were colleted after treatment for 24,48 and 72 hours.The morphological changes of cells were observed by the inverted phase-contrast microscopy,the proliferation-inhibitory rates of cells was detected by MTT method,the expression of HIF-1α at transcription level was detected by real-time fluorescent quantitative PCR.Results:The cell morphology was changed with increase of CoCl2 concentration and prolonging of treatment time of CoCl2.The MTT assay showed that the inhibitory rate of cell proliferation was enhanced also with increase of CoCl2 concentration and prolonging of treatment time of CoCl2 (r =0.435,r =0.389,P ＜ 0.05).The realtime fluorescent quantitative PCR showed that the mRNA expression of HIF-1α in K562 cells of hypoxia group was upregulated in concentration-and time-dependant manners.and displayed the positive correlation with concentration of CoCl2 and treatment time (r =0.954,r =0895).Conclusion:The hypoxia can inhibit the proliferation of K562 cells and up-requlate the expression of HIF-1α in cells,but does not induce cell differentiation.
摘要：
【目的】研究中度干旱（60％田間持水量）下，緩解辣椒幼苗干旱脅迫的適宜的CoCl2溶液濃度．【方法】以‘隴椒5號’辣椒為試材，采用不同濃度CoCl2（0．02、0．04、0．06、0．08、0．10、0．12 mmol／L）溶液澆灌處理，并測定處理期間辣椒幼苗相關(guān)生長生理指標．【結(jié)果】隨著CoCl2濃度的增加，辣椒幼苗株高、莖粗、地上部干質(zhì)量、根冠比呈先增大后減小的趨勢，均在0．06～0．08 mmol／L處理時出現(xiàn)峰值；CoCl2濃度在0．06 mmol／L以上時可顯著促進葉片中葉綠素含量的增加；0．06～0．10 mmol／L CoCl2處理辣椒幼苗葉片丙二醛和脯氨酸含量顯著低于單一干旱脅迫處理；高濃度的CoCl2（0．10～0．12 mmol／L）處理，對地下部干物質(zhì)的積累和根系的伸長有顯著的抑制作用．【結(jié)論】綜合各項生長及生理指標，外源施用0．06～0．08 mmol／L CoCl2可緩解辣椒幼苗干旱脅迫．%Obj ective]To determine suitable solution concentration of CoCl2 to reduce the moderate drought stress(60%field moisture capacity).[Method]‘LongJiao 5’was used as test material to be irriga-ted with the following concentrations of CoCl2 solution (0.02,0.04,0.06,0.08,0.10,0.12 mmol/L)and determined growth and physiological indicators of pepper seedlings during processing.[Result]With the increase of CoCl2 concentrations,plant height,stem diameter,shoot dry weight,shoot ratio of pepper seed-ling increased first then decreased,and the peak values all occurred at the concentration range of 0.06~0.08 mmol/L.CoCl2 concentration more than 0.06 mmol/L significantly promoted the increase of chloro-phyll content.MDA and proline content in pepper seedling leaves treated with 0.06~0.08 mmol/L CoCl2 than those under single drought stress (CK1 ).High concentrations of CoCl2 (0.10~0.12 mmol/L)signifi-cantly inhibited the underground dry matter accumulation and root elongation.[Conclusion]Based on com-prehensive analysis on various growth and physiological indicators,exogenous application of 0.06~0.08 mmol/L CoCl2 solution can alleviate drought stress on pepper seedlings.
摘要：
Objective To investigate the hypoxia injury and mechanism of Cobalt chloride ( CoCl2 ) on SH-SY5Y cells.Methods Different doses of CoCl2 were used to treat SH-SY5Y cells,MTT assay was employed to examine the growth of the cells .ELISA was used to detect the expressions of Bax ,Bcl-2 ,Caspase-3 and HIF-1α.Results The in-hibitory effect of CoCl 2 was appeared in a time and dosage dependent manner .CoCl 2 could increase the expression of Caspase-3, Bax and HIF-1α(P<0.05),while reduce the expression of Bcl-2 (P<0.01).Conclusion CoCl2 could induce hypoxia on SH-SY5 Y cells by up-regulating HIF-1αand inducing apoptosis .%目的：探討氯化鈷（ CoCl2）誘導(dǎo)SH-SY5Y細胞缺氧損傷的作用及相關(guān)機制。方法用四甲基偶氮唑藍比色法（ MTT）觀察不同濃度CoCl2對SH-SY5Y細胞生長情況影響；ELISA法檢測細胞內(nèi)相關(guān)凋亡蛋白Bcl-2、Bax、Caspase-3及缺氧相關(guān)蛋白HIF-1α的表達情況。結(jié)果 CoCl2可以誘導(dǎo)SH-SY5Y細胞損傷，并呈現(xiàn)濃度和時間依賴性；CoCl2作用于SH-Y5Y細胞導(dǎo)致HIF-1α、Caspace-3和Bax的表達增多（P＜0．05），Bcl-2表達減少（P＜0．01）。結(jié)論 CoCl2誘導(dǎo)SH-SY5Y細胞缺氧損傷與上調(diào)HIF-1α表達、促進細胞凋亡有關(guān)。
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