吗啡对HIV-1在MT2细胞和巨噬细胞中复制的影响
마배대HIV-1재MT2세포화거서세포중복제적영향
Effect of Morphine on the replication of HIV-1 in MT2 and macrophage
  • 万方数据
    中华检验医学杂志 중화검험의학잡지
    CHINESE JOURNAL OF LABORATORY MEDICINE
    2011年 7期 650-655 ,共6页

    梁冰玉%李敬云%庄道民%苏齐鉴%刘思扬%蒋俊俊%肖信%岑平%陈晖%梁浩

    량빙옥%리경운%장도민%소제감%류사양%장준준%초신%잠평%진휘%량호

    吗啡%HIV-1%HIV核心蛋白质p24%巨噬细胞%纳洛酮%病毒复制 嗎啡%HIV-1%HIV覈心蛋白質p24%巨噬細胞%納洛酮%病毒複製
    마배%HIV-1%HIV핵심단백질p24%거서세포%납락동%병독복제
    Morphine%HIV-1%HIV core protein p24%Macrophages%Naloxone%Virus replication
    目的 附测定双抗体夹心法检测p24抗原表达.用Student-t检验或方差分析(ANOVA)比较不同细胞处理组间p24抗原表达差异;采用重复测量数据的方差分析比较不同时间吗啡处理组比对照组的p24抗原表达增加或减少倍数的变化趋势.结果 HIV-1感染MT2细胞后第3、4、5和6天,3个浓度的吗啡处理的(1)组的p24抗原表达[第3天:(4.44±0.30)、(5.59±0.25)和(4.60±0.24) ng/ml;第4天:(24.30±2.66)、(31.73±1.17)和(26.02±0.37) ng/ml;第5天:(56.30±1.26)、(81.77±2.49)和(63.66±2.57) ng/ml;第6天:(150.70±8.97)、(243.09±8.93)和(173.72±7.73) ng/ml]均高于(4)组[第3~6天分别为(1.93±0.05)、(8.03±0.09)、(15.30±0.91)、(41.01±0.84) ng/ml],差异有统计学意义(第3天:t值分别为14.15、24.74和19.14,P均<0.01;第4天:t值分别为10.59、34.92和81.2,P均<0.01;第5天:t值分别为45.83、43.51和30.07,P均<0.01;第6天:t值分别为20.09、39.02和29.55,P均<0.01).3个浓度的吗啡处理的(1)组的p24抗原表达比(4)组增加的倍数,随HIV-1感染MT2细胞的时间的延长具有上升的趋势,时间效应具有统计学意义(F=842.18,P<0.01).HIV-1感染巨噬细胞组后第4、6、8和10天,3个浓度的吗啡处理的(5)组的p24抗原表达[第4天:(0.68±0.15)、(0.87±0.41)和(0.75±0.09) ng/ml;第6天:(1.64±0.57)、(2.07±0.12)和(1.75±0.17) ng/ml;第8天:(6.31±0.17)、(8.81±0.34)和(7.19±0.11) ng/ml;第10天:(32.30±17.55)、(50.74±17.55)和(39.74±0.56) ng/ml]均高于(8)组[第4、6、8、10天分别为(0.60±0.01)、(1.16±0.07)、(3.84±0.45)、(17.55±0.86) ng/ml],差异有统计学意义(第4天:t值分别为7.27、11.06和3.02,P均<0.05;第6天:t值分别为8.93、11.3和5.45,P均<0.01;第8天:t值分别为8.83、15.11和12.42,P均<0.01;第10天:t值分别为13.65、17.84和36.69,P均<0.01).3个浓度的吗啡处理的(5)组的p24抗原表达比(8)组增加的倍数,随HIV-1感染时间的延长具有上升的趋势,时间效应具有统计学意义(F=135.58,P<0.01).结论 吗啡能够促进HIV-1在MT2细胞和巨噬细胞内的复制,并且随着感染时间的延长而增加;吗啡促进HIV-1复制的作用可被纳洛酮阻断.
    목적 부측정쌍항체협심법검측p24항원표체.용Student-t검험혹방차분석(ANOVA)비교불동세포처리조간p24항원표체차이;채용중복측량수거적방차분석비교불동시간마배처리조비대조조적p24항원표체증가혹감소배수적변화추세.결과 HIV-1감염MT2세포후제3、4、5화6천,3개농도적마배처리적(1)조적p24항원표체[제3천:(4.44±0.30)、(5.59±0.25)화(4.60±0.24) ng/ml;제4천:(24.30±2.66)、(31.73±1.17)화(26.02±0.37) ng/ml;제5천:(56.30±1.26)、(81.77±2.49)화(63.66±2.57) ng/ml;제6천:(150.70±8.97)、(243.09±8.93)화(173.72±7.73) ng/ml]균고우(4)조[제3~6천분별위(1.93±0.05)、(8.03±0.09)、(15.30±0.91)、(41.01±0.84) ng/ml],차이유통계학의의(제3천:t치분별위14.15、24.74화19.14,P균<0.01;제4천:t치분별위10.59、34.92화81.2,P균<0.01;제5천:t치분별위45.83、43.51화30.07,P균<0.01;제6천:t치분별위20.09、39.02화29.55,P균<0.01).3개농도적마배처리적(1)조적p24항원표체비(4)조증가적배수,수HIV-1감염MT2세포적시간적연장구유상승적추세,시간효응구유통계학의의(F=842.18,P<0.01).HIV-1감염거서세포조후제4、6、8화10천,3개농도적마배처리적(5)조적p24항원표체[제4천:(0.68±0.15)、(0.87±0.41)화(0.75±0.09) ng/ml;제6천:(1.64±0.57)、(2.07±0.12)화(1.75±0.17) ng/ml;제8천:(6.31±0.17)、(8.81±0.34)화(7.19±0.11) ng/ml;제10천:(32.30±17.55)、(50.74±17.55)화(39.74±0.56) ng/ml]균고우(8)조[제4、6、8、10천분별위(0.60±0.01)、(1.16±0.07)、(3.84±0.45)、(17.55±0.86) ng/ml],차이유통계학의의(제4천:t치분별위7.27、11.06화3.02,P균<0.05;제6천:t치분별위8.93、11.3화5.45,P균<0.01;제8천:t치분별위8.83、15.11화12.42,P균<0.01;제10천:t치분별위13.65、17.84화36.69,P균<0.01).3개농도적마배처리적(5)조적p24항원표체비(8)조증가적배수,수HIV-1감염시간적연장구유상승적추세,시간효응구유통계학의의(F=135.58,P<0.01).결론 마배능구촉진HIV-1재MT2세포화거서세포내적복제,병차수착감염시간적연장이증가;마배촉진HIV-1복제적작용가피납락동조단.
    Objective To determine whether Morphine has the ability to enhance HIV-1 replication in MT2 and Macrophage in vitro and assess the influence of Naloxone on Morphine2s effect.Methods MT2 cells were randomly assigned into 4 groups: (1) Morphine treatment for MT2 group, (2) Morphine+Naloxone co-treatment for MT2 group, (3) Naloxone treatment for MT2 group and (4) MT2 Control;Macrophages were also randomly assigned into 4 groups: (5) Morphine treatment for Macrophage group, (6) Morphine+Naloxone co-treatment for Macrophage group, (7) Naloxone treatment for Macrophage group and (8) Macrophage Control. Group (2), (3), (6) and (7) were pre-treated with 10-8 mol/L Naloxone for 0.5 h, and then group (1) and (2) were treated with 10-12, 10-10 and 10-8 mol/L Morphine for 24 h;group (5) and (6) were disposed of 10-10 mol/L Morphine for 24 h.All 8 groups were added in HIV-1 viral strain with 50% tissue culture infective dose(TCID50).P24 antigen in MT2 cells culture supernatant at day 3, 4, 5 and 6, and in Macrophages culture supernatant at day 4, 6, 8, 10 and 12 after infection were determined with ELISA.Student2s t-test and ANOVA were used to compare the differential expression in different groups, and repeated measures ANOVA was used to compare the increasing or decreasing expression of p24 antigen in morphine treatment groups than that in the control group at different time points.Results On the 3rd day of infection with HIV-1 in MT2 cells, the expression of p24 antigen in 10-12, 10-10 and 10-8mol/L dose of group (1) were (4.44?.30), (5.59?.25) and (4.60?.24) ng/ml respectively, compared to control[(1.93?.05) ng/ml, t= 14.15, 24.74 and 19.14, all P<0.01].On the 4th day, 10-12, 10-10 and 10-8mol/L dose of group (1) resulted in a significant increase of p24 antigen expression [(24.30?.66), (31.73?.17) and (26.02?.37) ng/ml]in culture supernatants compared to control[(8.03?.09) ng/ml, t=10.59, 34.92 and 81.2, all P<0.01].On the 5th day, the expression of p24 antigen in 10-12, 10-10 and 10-8 mol/L dose of group (1) were (56.30?.26), (81.77?.49) and (63.66?.57) ng/ml respectively, compared to control [(15.30?.91) ng/ml, t= 45.83, 43.51 and 30.07, all P<0.01].On the 6th day, the expression of p24 antigen in 10-12, 10-10 and 10-8 mol/L dose of group (1) were (150.70?.97), (243.09?.93) and (173.72?.73) ng/ml respectively, compared to control [(41.01?.84) ng/ml, t= 21.09, 39.02 and 29.55, all P<0.01].The enhanced multiple of p24 antigen expression in three doses of morphine treatment group compared to control increased with HIV-1 infected MT2 cells time, trend analysis of repeated measurements showed statistically significant time effect (F=842.18, P<0.01). On the 4th day of infection with HIV-1 in Macrophage cells, the expression of p24 antigen in 10-12, 10-10 and 10-8 mol/L dose of group (5) were (0.68?.15), (0.87?.41) and (0.75?.09) ng/ml respectively, compared to control [(0.60?.01) ng/ml, t= 7.27, 11.06 and 3.02, all P<0.05]. On the 6th day, 10-12, 10-10 and 10-8 mol/L dose of group (5) resulted in a significant increase of p24 antigen expression[(1.64?.57) , (2.07?.12 ) and (1.75?.17) ng/ml]in culture supernatants compared to control [(1.16?.07) ng/ml, t=8.93, 11.3 and 5.45, all P<0.01].On the 8th day, the expression of p24 antigen in 10-12, 10-10 and 10-8 mol/L dose of group (5) were (6.31?.17), (8.81?.34) and (7.19?.11) ng/ml respectively, compared to control [(3.84?.45) ng/ml, t=8.83, 15.11 and 12.42, all P<0.01]. On the 10th day, the expression of p24 antigen in 10-12, 10-10 and 10-8 mol/L dose of Morphine treated group were (32.30?7.55), (50.74?7.55) and (39.74?.56) ng/ml respectively, compared to control [(17.55?.86) ng/ml, t= 13.65, 17.84 and 36.69, all P<0.01].The enhanced multiple of p24 antigen expression in three doses of group (5) compared to control increased with HIV-1 infected Macrophage cells time, trend analysis of repeated measurements showed statistically significant time effect (F=135.58, P<0.01).Conclusions Morphine has the ability to enhance HIV-1 replication in MT2 cell and Macrophage. This Morphine-mediated increase of p24 antigen expression can be blocked by Naloxone.
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