Objective and background： Although p21 ras has been reported to be upregulated in hepatocellular carcinoma complicating chronic hepatitis C type I, p21 ras has a different role in advanced stages, as it has been foun...Objective and background： Although p21 ras has been reported to be upregulated in hepatocellular carcinoma complicating chronic hepatitis C type I, p21 ras has a different role in advanced stages, as it has been found to be downregulated. The goal of this study was to investigate the status of p21 ras in early-stage/low-grade and late-stage/high-grade hepatocellular carcinoma and its possible link to apoptosis. Material and methods： Thirty-five cases each of chronic HCV hepatitis type 4 （group I） and cirrhosis with hepatocellular carcinoma （HCC） complicating chronic HCV hepatitis （groups Ⅱ and Ⅲ） were immunohistochemically evaluated using a p21 ras polyclonal antibody. The apoptotic index was determined in histologic sections using the terminal deoxynncleotidyl transferase-mediated d-UTP biotin nick end labeling （TUNEL） assay. Results： Significant differences （P=0.001） were detected in p21 ras protein expression between the three groups. A near 2-fold increase in p21 ras staining was observed in the cirrhotic cases compared to the hepatitis cases, and p21 ras expression was decreased in the HCC group, p21 ras expression correlated with stage （r=0.64, P--0.001） and grade （r=-0.65, P=0.001） in the HCC group and grade in the HCV group （r=0.44, P=0.008）. Both p21 ras expression and TUNEL-LI were significantly lower in large HCCs compared to small HCCs （P=0.01 each）. The TUNEL values were negatively correlated with stage in the HCC group （r=-0.85, P=0.001）. The TUNEL values were also negatively correlated with grade in both the HCV and HCC groups （r=0.89, P=0.001 and r=0.53, P=0.001, respectively）. The p21 ras scores were significantly correlated with the TUNEL-LI values in the HCC group （r=0.63, P=0.001） and HCV group （r=0.88, P=0.001）. Conclusions： p21 ras acts as an initiator in HCC complicating type 4 chronic HCV and is downregulated with HCC progression, which most likely promotes tumor cell survival because it facilitates the downregulation of apoptosis with tumor progression.展开更多
目的:探讨梗阻性胆管炎大鼠细胞免疫功能降低的发生机制及清热通下中药锦红片的影响.方法:♂SD大鼠24只建立急性梗阻性胆管炎模型,随机分为模型组(n=8)、锦红片治疗组(n=8)和单纯胆管梗阻组(n=8),检测血浆IL-2,CD_3^+,CD_4^+,CD_8^+,内...目的:探讨梗阻性胆管炎大鼠细胞免疫功能降低的发生机制及清热通下中药锦红片的影响.方法:♂SD大鼠24只建立急性梗阻性胆管炎模型,随机分为模型组(n=8)、锦红片治疗组(n=8)和单纯胆管梗阻组(n=8),检测血浆IL-2,CD_3^+,CD_4^+,CD_8^+,内毒素,胸腺指数,胸腺细胞凋亡指数及电镜下观察胸腺的超微结构及凋亡.结果:模型组IL-2,CD_3^+,CD_4^+和胸腺指数显著低于治疗组和单纯胆管梗阻组(IL-2:28.5±3.0 ng/L vs 33.9±3.6 ng/L,39.6±2.2 ng/L,P<0.05,P<0.01;CD_3^+:54.5%±5.5% vs 70.7%±4.8%,66.3%±7.1%,均P<0.01;CD_4^+:34.5%±8.3% vs 44.2%±3.3%,44.5%±4.2%,均P<0.01:胸腺指数:0.89±0.18 vs 1.10±0.13.1.12±0.24,均P<0.05),CD_8^+3组间没有统计学差异,血浆内毒素和凋亡指数明显高于治疗组和梗阻组(内毒素:0.85±0.14 Eu/mL vs 0.53±0.10 EU/mL,0.49±0.11 EU/mL,均P<0.01;凋亡指数:25.7±5.1 vs 15.8±5.5.9.0±3.1.P<0.05,P<0.01),模型组胸腺可见较多典型的凋亡细胞,结果显示经中药干预治疗后,免疫功能、内毒素血症和胸腺细胞凋亡有所改善,接近单纯胆管梗阻组水平.结论:梗阻性胆管炎大鼠存在免疫功能降低,胸腺细胞异常凋亡.锦红片对维持免疫机能的稳定有积极的意义.展开更多
目的:研究缺血再灌流时异丙酚对肠上皮细胞凋亡的影响及可能机制.方法:96只成年♂Wistar大鼠,随机分为假手术组、缺血再灌流+生理盐水组(I/R+NS)和I/R+异丙酚组(I/R+Pr).采用夹闭肠系膜上动脉(SMA)的方法制作肠缺血再灌流模型.以上各组...目的:研究缺血再灌流时异丙酚对肠上皮细胞凋亡的影响及可能机制.方法:96只成年♂Wistar大鼠,随机分为假手术组、缺血再灌流+生理盐水组(I/R+NS)和I/R+异丙酚组(I/R+Pr).采用夹闭肠系膜上动脉(SMA)的方法制作肠缺血再灌流模型.以上各组分别在再灌流后0,30,60,120和240 min(每时间点8只)处死动物取肠袋组织.采用病理学方法观察肠上皮细胞损伤指数;原位DNA末端标记法(TUNEL)检测肠上皮细胞凋亡率的变化;免疫组化法检测肠上皮细胞Caspase-3,bcl-2表达的变化.结果:I/R+Pr组与I/R+NS组相比,肠上皮细胞病理变化较轻,肠上皮细胞的凋亡率明显下降(P<0.01),再灌流后0,30,60,120和240 min肠上皮细胞中Caspase-3阳性细胞数明显减少(104.4±5.3 vs 146.4±7.6;97.4±6.2 vs 130.4±7.4;134.4±5.1 vs 170.4±8.1;125.4±6.2 vs 160.4±9.5:101±5.8 vs 120.4±8.2,均P<0.01),而bcl-2阳性细胞数明显增加(13.34±4.12 vs 6.72±2.59;14.96±4.85 vs 8.24±3.13;15.29±5.28 vs 9.63±2.89;10.39±3.61 vs 9.63±2.89;10.39±3.61 vs 5.96±1.93;11.08±4.83 vs 6.87±2.43,均P<0.01).结论:异丙酚能抑制缺血再灌流时肠上皮细胞Caspase-3表达,而增加bcl-2表达,减少肠上皮细胞的凋亡.展开更多
目的:观察大鼠酒精性肝病组织病理形态学改变,探讨细胞凋亡与细胞色素P4502E1的表达以及和氧化应激的关系.方法:用酒精灌胃法制备酒精性肝病大鼠模型,模型组给予酒精8 g/kg,每天分2次灌胃连续8 wk,对照组给予等量的生理盐水灌胃.实验8 w...目的:观察大鼠酒精性肝病组织病理形态学改变,探讨细胞凋亡与细胞色素P4502E1的表达以及和氧化应激的关系.方法:用酒精灌胃法制备酒精性肝病大鼠模型,模型组给予酒精8 g/kg,每天分2次灌胃连续8 wk,对照组给予等量的生理盐水灌胃.实验8 wk末,观察肝组织的病理形态学改变,用原位末端标记法(TUNEL)检测肝细胞凋亡,用免疫组化法检测肝组织中Caspase-3蛋白表达,用全自动生化仪检测ALT和AST的含量,用PCR法测定肝细胞色素P4502E1的基因表达,分别用硫代巴比妥酸法(TBA法)和黄嘌呤氧化酶法测定肝组织丙二醛(MDA)的含量和超氧化物歧化酶(SOD)的活力.结果:模型组凋亡的肝细胞明显增多,主要分布在中央静脉周围、点状和灶状坏死区;Caspase-3主要分布于中央静脉及肝细胞坏死灶周围细胞的胞质中.模型组肝细胞凋亡指数(AI)和Caspase-3蛋白表达强度明显高于对照组(AI:6.2%±1.7% vs 1.7%±0.8%;Caspase-3:66.7% vs 9.5%,P<0.05,P<0.01).CYP2E1表达:对照组c1基因频率为91.6%,c2基因频率为8.4%;模型组c1基因频率为53.4%,c2基因频率为46.6%,均有显著性差异(P<0.05).长期酒精摄入大鼠血清MDA含量增加(41.53±7.43μmol/L vs 15.72±2.06μmol/L,P<0.05),SOD活力下降(353.12±61.02 kU/L vs 636.82±138.60 kU/L,P<0.05),与酒精性肝病肝细胞凋亡程度有相关性(r=0.644,r=-0.511).结论:长期酒精摄入可引起大鼠酒精性肝病及及肝功能损伤,肝细胞凋亡明显增加.CYP2E1基因PstⅠ及RsaⅠRFLPs与酒精性肝病有关,其中c2基因可能与大鼠酒精性肝病的发生有关.MDA含量和SOD活力在酒精性肝病的肝细胞凋亡过程及脂质过氧化反应中发挥重要作用.展开更多
目的了解新生期注射己烯雌酚(diethylstilbestrol,DES)对雌性BALB/c小鼠生后不同时期脾细胞凋亡及Bcl-2、Bax的影响。方法新生雌性BALB/c小鼠,在生后24h内颈背部皮下注射DES40μg,每隔24h注射一次,共连续5次,对照组平行注射等量无菌豆...目的了解新生期注射己烯雌酚(diethylstilbestrol,DES)对雌性BALB/c小鼠生后不同时期脾细胞凋亡及Bcl-2、Bax的影响。方法新生雌性BALB/c小鼠,在生后24h内颈背部皮下注射DES40μg,每隔24h注射一次,共连续5次,对照组平行注射等量无菌豆油。分别在生后7、14、21、35和49d将小鼠处死,取其脾进行常规石蜡包埋和切片,进行TUNEL(TdT-mediated dUTP nick endlabeling,)检测以及Bcl-2、Bax的免疫组织化学检测。结果与对照组相比,DES组雌性BALB/c小鼠脾细胞的TUNEL阳性细胞数目在生后各点均有显著性增加。Bcl-2在各时间点DES组的表达均弱于相应的对照组。Bax在生后7d的对照组和DES组中的表达差异无统计学意义,而在生后14、21、35和49d DES组的阳性表达明显多于对照组。结论新生期注射DES可以导致雌性BALB/c小鼠脾细胞凋亡明显增加。细胞凋亡在对照组和DES组不同时间点的动态变化以及在两组间相同时间点的差异可能与Bcl-2表达的减少以及Bax表达的增加有关。展开更多
文摘Objective and background： Although p21 ras has been reported to be upregulated in hepatocellular carcinoma complicating chronic hepatitis C type I, p21 ras has a different role in advanced stages, as it has been found to be downregulated. The goal of this study was to investigate the status of p21 ras in early-stage/low-grade and late-stage/high-grade hepatocellular carcinoma and its possible link to apoptosis. Material and methods： Thirty-five cases each of chronic HCV hepatitis type 4 （group I） and cirrhosis with hepatocellular carcinoma （HCC） complicating chronic HCV hepatitis （groups Ⅱ and Ⅲ） were immunohistochemically evaluated using a p21 ras polyclonal antibody. The apoptotic index was determined in histologic sections using the terminal deoxynncleotidyl transferase-mediated d-UTP biotin nick end labeling （TUNEL） assay. Results： Significant differences （P=0.001） were detected in p21 ras protein expression between the three groups. A near 2-fold increase in p21 ras staining was observed in the cirrhotic cases compared to the hepatitis cases, and p21 ras expression was decreased in the HCC group, p21 ras expression correlated with stage （r=0.64, P--0.001） and grade （r=-0.65, P=0.001） in the HCC group and grade in the HCV group （r=0.44, P=0.008）. Both p21 ras expression and TUNEL-LI were significantly lower in large HCCs compared to small HCCs （P=0.01 each）. The TUNEL values were negatively correlated with stage in the HCC group （r=-0.85, P=0.001）. The TUNEL values were also negatively correlated with grade in both the HCV and HCC groups （r=0.89, P=0.001 and r=0.53, P=0.001, respectively）. The p21 ras scores were significantly correlated with the TUNEL-LI values in the HCC group （r=0.63, P=0.001） and HCV group （r=0.88, P=0.001）. Conclusions： p21 ras acts as an initiator in HCC complicating type 4 chronic HCV and is downregulated with HCC progression, which most likely promotes tumor cell survival because it facilitates the downregulation of apoptosis with tumor progression.
文摘目的:探讨梗阻性胆管炎大鼠细胞免疫功能降低的发生机制及清热通下中药锦红片的影响.方法:♂SD大鼠24只建立急性梗阻性胆管炎模型,随机分为模型组(n=8)、锦红片治疗组(n=8)和单纯胆管梗阻组(n=8),检测血浆IL-2,CD_3^+,CD_4^+,CD_8^+,内毒素,胸腺指数,胸腺细胞凋亡指数及电镜下观察胸腺的超微结构及凋亡.结果:模型组IL-2,CD_3^+,CD_4^+和胸腺指数显著低于治疗组和单纯胆管梗阻组(IL-2:28.5±3.0 ng/L vs 33.9±3.6 ng/L,39.6±2.2 ng/L,P<0.05,P<0.01;CD_3^+:54.5%±5.5% vs 70.7%±4.8%,66.3%±7.1%,均P<0.01;CD_4^+:34.5%±8.3% vs 44.2%±3.3%,44.5%±4.2%,均P<0.01:胸腺指数:0.89±0.18 vs 1.10±0.13.1.12±0.24,均P<0.05),CD_8^+3组间没有统计学差异,血浆内毒素和凋亡指数明显高于治疗组和梗阻组(内毒素:0.85±0.14 Eu/mL vs 0.53±0.10 EU/mL,0.49±0.11 EU/mL,均P<0.01;凋亡指数:25.7±5.1 vs 15.8±5.5.9.0±3.1.P<0.05,P<0.01),模型组胸腺可见较多典型的凋亡细胞,结果显示经中药干预治疗后,免疫功能、内毒素血症和胸腺细胞凋亡有所改善,接近单纯胆管梗阻组水平.结论:梗阻性胆管炎大鼠存在免疫功能降低,胸腺细胞异常凋亡.锦红片对维持免疫机能的稳定有积极的意义.
文摘目的:研究缺血再灌流时异丙酚对肠上皮细胞凋亡的影响及可能机制.方法:96只成年♂Wistar大鼠,随机分为假手术组、缺血再灌流+生理盐水组(I/R+NS)和I/R+异丙酚组(I/R+Pr).采用夹闭肠系膜上动脉(SMA)的方法制作肠缺血再灌流模型.以上各组分别在再灌流后0,30,60,120和240 min(每时间点8只)处死动物取肠袋组织.采用病理学方法观察肠上皮细胞损伤指数;原位DNA末端标记法(TUNEL)检测肠上皮细胞凋亡率的变化;免疫组化法检测肠上皮细胞Caspase-3,bcl-2表达的变化.结果:I/R+Pr组与I/R+NS组相比,肠上皮细胞病理变化较轻,肠上皮细胞的凋亡率明显下降(P<0.01),再灌流后0,30,60,120和240 min肠上皮细胞中Caspase-3阳性细胞数明显减少(104.4±5.3 vs 146.4±7.6;97.4±6.2 vs 130.4±7.4;134.4±5.1 vs 170.4±8.1;125.4±6.2 vs 160.4±9.5:101±5.8 vs 120.4±8.2,均P<0.01),而bcl-2阳性细胞数明显增加(13.34±4.12 vs 6.72±2.59;14.96±4.85 vs 8.24±3.13;15.29±5.28 vs 9.63±2.89;10.39±3.61 vs 9.63±2.89;10.39±3.61 vs 5.96±1.93;11.08±4.83 vs 6.87±2.43,均P<0.01).结论:异丙酚能抑制缺血再灌流时肠上皮细胞Caspase-3表达,而增加bcl-2表达,减少肠上皮细胞的凋亡.
文摘目的:观察大鼠酒精性肝病组织病理形态学改变,探讨细胞凋亡与细胞色素P4502E1的表达以及和氧化应激的关系.方法:用酒精灌胃法制备酒精性肝病大鼠模型,模型组给予酒精8 g/kg,每天分2次灌胃连续8 wk,对照组给予等量的生理盐水灌胃.实验8 wk末,观察肝组织的病理形态学改变,用原位末端标记法(TUNEL)检测肝细胞凋亡,用免疫组化法检测肝组织中Caspase-3蛋白表达,用全自动生化仪检测ALT和AST的含量,用PCR法测定肝细胞色素P4502E1的基因表达,分别用硫代巴比妥酸法(TBA法)和黄嘌呤氧化酶法测定肝组织丙二醛(MDA)的含量和超氧化物歧化酶(SOD)的活力.结果:模型组凋亡的肝细胞明显增多,主要分布在中央静脉周围、点状和灶状坏死区;Caspase-3主要分布于中央静脉及肝细胞坏死灶周围细胞的胞质中.模型组肝细胞凋亡指数(AI)和Caspase-3蛋白表达强度明显高于对照组(AI:6.2%±1.7% vs 1.7%±0.8%;Caspase-3:66.7% vs 9.5%,P<0.05,P<0.01).CYP2E1表达:对照组c1基因频率为91.6%,c2基因频率为8.4%;模型组c1基因频率为53.4%,c2基因频率为46.6%,均有显著性差异(P<0.05).长期酒精摄入大鼠血清MDA含量增加(41.53±7.43μmol/L vs 15.72±2.06μmol/L,P<0.05),SOD活力下降(353.12±61.02 kU/L vs 636.82±138.60 kU/L,P<0.05),与酒精性肝病肝细胞凋亡程度有相关性(r=0.644,r=-0.511).结论:长期酒精摄入可引起大鼠酒精性肝病及及肝功能损伤,肝细胞凋亡明显增加.CYP2E1基因PstⅠ及RsaⅠRFLPs与酒精性肝病有关,其中c2基因可能与大鼠酒精性肝病的发生有关.MDA含量和SOD活力在酒精性肝病的肝细胞凋亡过程及脂质过氧化反应中发挥重要作用.
文摘目的了解新生期注射己烯雌酚(diethylstilbestrol,DES)对雌性BALB/c小鼠生后不同时期脾细胞凋亡及Bcl-2、Bax的影响。方法新生雌性BALB/c小鼠,在生后24h内颈背部皮下注射DES40μg,每隔24h注射一次,共连续5次,对照组平行注射等量无菌豆油。分别在生后7、14、21、35和49d将小鼠处死,取其脾进行常规石蜡包埋和切片,进行TUNEL(TdT-mediated dUTP nick endlabeling,)检测以及Bcl-2、Bax的免疫组织化学检测。结果与对照组相比,DES组雌性BALB/c小鼠脾细胞的TUNEL阳性细胞数目在生后各点均有显著性增加。Bcl-2在各时间点DES组的表达均弱于相应的对照组。Bax在生后7d的对照组和DES组中的表达差异无统计学意义,而在生后14、21、35和49d DES组的阳性表达明显多于对照组。结论新生期注射DES可以导致雌性BALB/c小鼠脾细胞凋亡明显增加。细胞凋亡在对照组和DES组不同时间点的动态变化以及在两组间相同时间点的差异可能与Bcl-2表达的减少以及Bax表达的增加有关。
