中华航海医学与高气压医学杂志
中華航海醫學與高氣壓醫學雜誌
중화항해의학여고기압의학잡지
CHINESE JOURNAL OF NAUTICAL MEDICINE AND HYPERBARIC MEDICINE
2014年
4期
242-246
,共5页
郑洁%吴刚%胡国信%彭雁忠%熊晓佳
鄭潔%吳剛%鬍國信%彭雁忠%熊曉佳
정길%오강%호국신%팽안충%웅효가
异甘草酸镁%肝细胞%缺血再灌注
異甘草痠鎂%肝細胞%缺血再灌註
이감초산미%간세포%결혈재관주
Magnesium isoglycyrrhizinate%Hepatocyte%Hypoxia,reperfusion
目的 观察异甘草酸镁(magnesium isoglycyrrhizinate,MgIG)对大鼠肝细胞模拟缺血再灌注损伤的细胞凋亡影响及其机制.方法 分离、培养、鉴定大鼠肝细胞,将对数生长期肝细胞,按照数字表法随机分为模拟缺血再灌注损伤模型组和MgIG低浓度组(0.1 g/L)、中浓度组(1g/L)、高浓度组(10 g/L),预处理24 h后,模拟缺氧6h、复氧4h,进行以下实验观察:(1)应用光镜观察细胞的形态改变;(2)应用四甲基偶氮唑盐(MTT)法检测MgIG对肝细胞活力的影响;(3)应用吖啶橙/嗅化乙啶(AO/EB)染色观察细胞的凋亡情况,计算细胞凋亡率;(4)应用RT-qPCR法检测各组bcl-2、bax基因的表达量.结果 (1)MTT法结果表明,一定浓度的MgIG (0.10、1、10 g/L)能提高缺氧-复氧损伤的肝细胞A值(0.285 ±0.017、0.320±0.008、0.341 ±0.019),与模型组比较,差异有统计学意义(P＜0.01);(2)AO/EB染色结果显示,低、中、高浓度组细胞凋亡率(0.5763±0.0405、0.2866±0.0619、0.1261±0.0439)分别低于对照组(0.7124 ±0.0581),差异有统计学意义(P＜0.05);(3)RT-qPCR结果显示低、中、高浓度组bcl-2基因mRNA的表达量(3.350±0.151、9.279±0.350、15.290±0.756)分别高于对照组(0.928±0.068),差异有统计学意义(P＜0.01),低、中、高浓度组Bax mRNA的表达量(0.743±0.035、0.265 ±0.065、0.119 ±0.019)低于对照组(0.945±0.063)差异有统计学意义(P＜0.01).结论 MgIG能提高缺血再灌注损伤的肝细胞活力和降低肝细胞凋亡,作用机制可能与提高bcl-2的mRNA表达、降低bax mRNA的表达有关.
目的 觀察異甘草痠鎂(magnesium isoglycyrrhizinate,MgIG)對大鼠肝細胞模擬缺血再灌註損傷的細胞凋亡影響及其機製.方法 分離、培養、鑒定大鼠肝細胞,將對數生長期肝細胞,按照數字錶法隨機分為模擬缺血再灌註損傷模型組和MgIG低濃度組(0.1 g/L)、中濃度組(1g/L)、高濃度組(10 g/L),預處理24 h後,模擬缺氧6h、複氧4h,進行以下實驗觀察:(1)應用光鏡觀察細胞的形態改變;(2)應用四甲基偶氮唑鹽(MTT)法檢測MgIG對肝細胞活力的影響;(3)應用吖啶橙/嗅化乙啶(AO/EB)染色觀察細胞的凋亡情況,計算細胞凋亡率;(4)應用RT-qPCR法檢測各組bcl-2、bax基因的錶達量.結果 (1)MTT法結果錶明,一定濃度的MgIG (0.10、1、10 g/L)能提高缺氧-複氧損傷的肝細胞A值(0.285 ±0.017、0.320±0.008、0.341 ±0.019),與模型組比較,差異有統計學意義(P＜0.01);(2)AO/EB染色結果顯示,低、中、高濃度組細胞凋亡率(0.5763±0.0405、0.2866±0.0619、0.1261±0.0439)分彆低于對照組(0.7124 ±0.0581),差異有統計學意義(P＜0.05);(3)RT-qPCR結果顯示低、中、高濃度組bcl-2基因mRNA的錶達量(3.350±0.151、9.279±0.350、15.290±0.756)分彆高于對照組(0.928±0.068),差異有統計學意義(P＜0.01),低、中、高濃度組Bax mRNA的錶達量(0.743±0.035、0.265 ±0.065、0.119 ±0.019)低于對照組(0.945±0.063)差異有統計學意義(P＜0.01).結論 MgIG能提高缺血再灌註損傷的肝細胞活力和降低肝細胞凋亡,作用機製可能與提高bcl-2的mRNA錶達、降低bax mRNA的錶達有關.
목적 관찰이감초산미(magnesium isoglycyrrhizinate,MgIG)대대서간세포모의결혈재관주손상적세포조망영향급기궤제.방법 분리、배양、감정대서간세포,장대수생장기간세포,안조수자표법수궤분위모의결혈재관주손상모형조화MgIG저농도조(0.1 g/L)、중농도조(1g/L)、고농도조(10 g/L),예처리24 h후,모의결양6h、복양4h,진행이하실험관찰:(1)응용광경관찰세포적형태개변;(2)응용사갑기우담서염(MTT)법검측MgIG대간세포활력적영향;(3)응용아정등/후화을정(AO/EB)염색관찰세포적조망정황,계산세포조망솔;(4)응용RT-qPCR법검측각조bcl-2、bax기인적표체량.결과 (1)MTT법결과표명,일정농도적MgIG (0.10、1、10 g/L)능제고결양-복양손상적간세포A치(0.285 ±0.017、0.320±0.008、0.341 ±0.019),여모형조비교,차이유통계학의의(P＜0.01);(2)AO/EB염색결과현시,저、중、고농도조세포조망솔(0.5763±0.0405、0.2866±0.0619、0.1261±0.0439)분별저우대조조(0.7124 ±0.0581),차이유통계학의의(P＜0.05);(3)RT-qPCR결과현시저、중、고농도조bcl-2기인mRNA적표체량(3.350±0.151、9.279±0.350、15.290±0.756)분별고우대조조(0.928±0.068),차이유통계학의의(P＜0.01),저、중、고농도조Bax mRNA적표체량(0.743±0.035、0.265 ±0.065、0.119 ±0.019)저우대조조(0.945±0.063)차이유통계학의의(P＜0.01).결론 MgIG능제고결혈재관주손상적간세포활력화강저간세포조망,작용궤제가능여제고bcl-2적mRNA표체、강저bax mRNA적표체유관.
Objective To observe the effects of magnesium isoglycyrrhizinate (MgIG) on the apoptosis of hepatocytes induced by simulated hypoxia reperfusion in rats and related mechanism.Methods Hepatocytes of the rats were isolated,cultured and identified,then,the liver cells in logarithmic growth phase were randomly divided into 4 groups:the hypoxia reperfusion injury model group,the low MgIG group (0.1 g/L),the moderate MgIG group (1 g/L) and the high MgIG group(10 g/L).Twenty-four hours after pre-treatment (exposure to simulated hypoxia for 6 h and re-perfusion for 4 h),following measurements were performed.① Changes in cell morphology were observed by optical microscopy.② Effects of MgIG on the vitality of hepatocytes were detected by MTT.③Apoptosis and rate of apoptosis were detected and calculated by AO/EB.④The expression levels of bcl-2 and Bax in all the groups were monitored by real time quantitative fluorescence PCR (RT-qPCR).Then,the detected data were analyzed by SPSS17.0.T test was applied for the comparison of the two mean sample lots,and single-factor ANOVA was used for the comparison of multiple samples.Results ①MTT assay indicated that MgIG at some concentrations (0.1,1 and 10 g/L)could improve the A value of the damaged hepatocytes induced by ischemia and re-perfusion(0.285 ± 0.017,0.320 ± 0.008 and 0.341 ± 0.019),as compared with that of the model group,with statistical significance (P ＜ 0.01).②AO/EB staining revealed that apoptosis rates in the low,moderate and high MgIG groups were(0.5763 ± 0.0405,0.2866 ±0.0619 and 0.1261 ±0.0439),which were all lower than that of the control group (0.7124 ± 0.0581),also with statistical significance (P ＜ 0.05).③RT-qPCR monitoring indicated that the expression levels of bcl-2 mRNA in the low,moderate and high MgIG groups (3.350 ± 0.151,9.279 ± 0.350 and 15.290±0.756) were all higher than that of the control group (0.928 ± 0.068),also with statistical significance(P ＜0.01).The expression levels of Bax mRNA in the low,moderate and high MgIG groups (0.743 ± 0.035,0.265 ± 0.065 and 0.119 ± 0.019)were all lower than that of the control group (0.945 ± 0.063),with statistical significance as well (P ＜ 0.01)Conclusions MgIG could increase the vitality of damaged hepatocytes induced by hypoxia-reperfusion and decrease rate of apoptosis,the mechanism of which might be associated with higher expression of bcl-2 mRNA and lower expression of Bax mRNA.
