中国地方病学杂志
中國地方病學雜誌
중국지방병학잡지
CHINESE JOURNAL OF ENDEMIOLOGY
2010年
1期
9-12
,共4页
马艳%孔席丽%郑玉建%吴顺华%张杰
馬豔%孔席麗%鄭玉建%吳順華%張傑
마염%공석려%정옥건%오순화%장걸
砷%金属硫蛋白%基因表达%淋巴细胞
砷%金屬硫蛋白%基因錶達%淋巴細胞
신%금속류단백%기인표체%림파세포
Arsenic%Metallothionein%Gene expression%Lymphocytes
目的 探讨不同剂量亚砷酸钠对体外培养人淋巴细胞金属硫蛋白(MT)基因表达的影响及MT表达与细胞存活率的相互关系.方法 无菌抽取健康人血液,分离出淋巴细胞,分别用0(对照)、2、5、10、15、30、60μmol/L亚砷酸钠作用于淋巴细胞,于24、48、72 h后,以四噻唑蓝比色法测定细胞存活率;染毒72 h后采用RT-PCR法检测MT-1、MT-2基因表达情况.结果 2、5μmol/L染毒组细胞存活率[(115.50±11.80)%、(130.49±8.28)%]高于对照组[(100.00±0.00)%,P均<0.05];10、15、30、60μmol/L染毒组细胞存活率[(78.12±9.33)%、(71.62±10.82)%、(52.06±3.05)%、(40.98±5.41)%]随染毒剂量增高而逐渐降低,与对照组及2、5μmol/L组比较差异有统计学意义(P均<0.05).不同剂量亚砷酸钠作用72 h,对照组及2、5、10、15、30、60μmol/L染毒组MT-1基因表达(0.925±0.123、1.082±0.504、1.103±0.170、0.927±0.056、0.730±0.307、0.604±0.173、0.540±0.075)组间比较差异无统计学意义(P均>0.05);2、5 μmol/L染毒组(1.503±0.212、1.557±0.377)与对照组(0.762±0.112)比较MT-2表达上调(P均<0.05),且较其他各组表达增加(P均<0.05),而10、15、30、60μmoL/L染毒组MT-2表达(0.814±0.139、0.679±0.201、0.607±0.229、0.533±0.102)呈下降趋势,但组间比较差异无统计学意义(P均>0.05).MT-1、MT-2表达水平与细胞存活率之间存在正相关关系(r值分别为0.955、0.909,P均<0.05).结论 10 μmol/L以上剂量的砷可能对淋巴细胞MT表达有抑制作用,而低剂量(2、5μmol/L)砷可能刺激淋巴细胞的MT-2表达上调,提高细胞MT水平,起到保护细胞、提高细胞存活率的作用.
目的 探討不同劑量亞砷痠鈉對體外培養人淋巴細胞金屬硫蛋白(MT)基因錶達的影響及MT錶達與細胞存活率的相互關繫.方法 無菌抽取健康人血液,分離齣淋巴細胞,分彆用0(對照)、2、5、10、15、30、60μmol/L亞砷痠鈉作用于淋巴細胞,于24、48、72 h後,以四噻唑藍比色法測定細胞存活率;染毒72 h後採用RT-PCR法檢測MT-1、MT-2基因錶達情況.結果 2、5μmol/L染毒組細胞存活率[(115.50±11.80)%、(130.49±8.28)%]高于對照組[(100.00±0.00)%,P均<0.05];10、15、30、60μmol/L染毒組細胞存活率[(78.12±9.33)%、(71.62±10.82)%、(52.06±3.05)%、(40.98±5.41)%]隨染毒劑量增高而逐漸降低,與對照組及2、5μmol/L組比較差異有統計學意義(P均<0.05).不同劑量亞砷痠鈉作用72 h,對照組及2、5、10、15、30、60μmol/L染毒組MT-1基因錶達(0.925±0.123、1.082±0.504、1.103±0.170、0.927±0.056、0.730±0.307、0.604±0.173、0.540±0.075)組間比較差異無統計學意義(P均>0.05);2、5 μmol/L染毒組(1.503±0.212、1.557±0.377)與對照組(0.762±0.112)比較MT-2錶達上調(P均<0.05),且較其他各組錶達增加(P均<0.05),而10、15、30、60μmoL/L染毒組MT-2錶達(0.814±0.139、0.679±0.201、0.607±0.229、0.533±0.102)呈下降趨勢,但組間比較差異無統計學意義(P均>0.05).MT-1、MT-2錶達水平與細胞存活率之間存在正相關關繫(r值分彆為0.955、0.909,P均<0.05).結論 10 μmol/L以上劑量的砷可能對淋巴細胞MT錶達有抑製作用,而低劑量(2、5μmol/L)砷可能刺激淋巴細胞的MT-2錶達上調,提高細胞MT水平,起到保護細胞、提高細胞存活率的作用.
목적 탐토불동제량아신산납대체외배양인림파세포금속류단백(MT)기인표체적영향급MT표체여세포존활솔적상호관계.방법 무균추취건강인혈액,분리출림파세포,분별용0(대조)、2、5、10、15、30、60μmol/L아신산납작용우림파세포,우24、48、72 h후,이사새서람비색법측정세포존활솔;염독72 h후채용RT-PCR법검측MT-1、MT-2기인표체정황.결과 2、5μmol/L염독조세포존활솔[(115.50±11.80)%、(130.49±8.28)%]고우대조조[(100.00±0.00)%,P균<0.05];10、15、30、60μmol/L염독조세포존활솔[(78.12±9.33)%、(71.62±10.82)%、(52.06±3.05)%、(40.98±5.41)%]수염독제량증고이축점강저,여대조조급2、5μmol/L조비교차이유통계학의의(P균<0.05).불동제량아신산납작용72 h,대조조급2、5、10、15、30、60μmol/L염독조MT-1기인표체(0.925±0.123、1.082±0.504、1.103±0.170、0.927±0.056、0.730±0.307、0.604±0.173、0.540±0.075)조간비교차이무통계학의의(P균>0.05);2、5 μmol/L염독조(1.503±0.212、1.557±0.377)여대조조(0.762±0.112)비교MT-2표체상조(P균<0.05),차교기타각조표체증가(P균<0.05),이10、15、30、60μmoL/L염독조MT-2표체(0.814±0.139、0.679±0.201、0.607±0.229、0.533±0.102)정하강추세,단조간비교차이무통계학의의(P균>0.05).MT-1、MT-2표체수평여세포존활솔지간존재정상관관계(r치분별위0.955、0.909,P균<0.05).결론 10 μmol/L이상제량적신가능대림파세포MT표체유억제작용,이저제량(2、5μmol/L)신가능자격림파세포적MT-2표체상조,제고세포MT수평,기도보호세포、제고세포존활솔적작용.
Objective To explore the effects of sodium arsenie on Metallothionein(MT) isoforms genes expression,and to study the relevance between the MT expression and cell survival percentage. Methods Healthy persons blood was extracted aseptically and the lymphocytes were separated. The lymphocytes were treated by 0 (control) ,2,5,10,15,30,60 μmol/L sodium arsenites, respectively. The cell survival percentage was determined by methyl thiazolyl tetrazolium(MTT) reduction assay at 24,48,72 h intervals, while the expression of MT-1 and MT-2 were examined by RT-PCR in 72 h. Results The cell survival percentage in 2,5μmoL/L groups were (115.50± 11.80)% and (130.49±8.28)%,which were all higher than those in the control group [(100.00±0.00)%,all P < 0.05]at any intervals. In 10,15,30,60 μmol/L groups,the cell survival percentage[(78.12±9.33)%,(71.62± 10.82)%,(52.06±3.05)%,(40.98±5.41)%]increased along with the decrease of concentration,which showed a significant difference between these groups and the control group and 2,5μmol/L groups(all P < 0.05). There was no significant difference on MT-1 expression after exposed to different concentration sodium arsenites (0,2,5,10,15,30,60 μmol/L) for72 h(0.925±0.123,1.082±0.504,1.103±0.170,0.927±0.056,0.730±0.307,0.604± 0.173,0.540±0.075,all P > 0.05). The expression of MT-2 in 2,5μmol/L groups(1.503±0.212,1.557±0.377) was up regulated compared with that in control group(0.702±0.112) and MT-2 also expressed more than that in other groups(all P < 0.05). However,the expression of MT-2 declined when the concentraion was 10,15,30,60 μmol/L(0.814±0.139,0.679±0.201,0.607±0.229,0.533±0.102). There was no significant difference among the groups (all P > 0.05). The expression level oE MT-1,MT-2 was positively correlated with the cell survival pereentage(r = 0.955,0.909,all P < 0.05). Conclusions The sodium arsenite at concentration of 10 μmoL/L might inhibit the expression of MT of lymphocytes and low concentration sodium arsenite (2,5 μmol/L) might stimulate the lymphocytes to regulate the expression of MT-2 to higher levels,which can increase the cell survival percentage and exert the function of protecting cells.
