腹腔引流灌洗时机对大鼠重症急性胰腺炎的影响及作用机制

目的 拟在大鼠重症急性胰腺炎（SAP）模型中探讨腹腔引流灌洗时机对病程转归的影响并探究其作用机制。方法 清洁级SD大鼠,随机分为SAP模型组、早期引流组、延期引流组、早期灌洗组和延期灌洗组。比较各组24 h后大鼠生存率,检测大鼠血清和腹水中IL-1β、IL-6、IL-8、IL-10和TNF-α的浓度。结果 ①与模型组比较,早期引流组和早期灌洗组生存率显著提高（P＜0.01 ）,而延期引流组和延期灌洗组与模型组差异无统计学意义（P＞0.05）。②与模型组比较,早期引流组血清TNF-α浓度显著下降（P＜0.05）;与早期引流组比较,早期灌洗组血清IL-8浓度下降、IL-10浓度升高（P＜0.05）。③与模型组比较,早期引流组和早期灌洗组腹水中IL-1β、IL-6、和TNF-α水平显著降低（P＜0.05）,且早期灌洗组腹水中IL-10浓度升高（P＜0.05）;延期引流组中仅腹水TNF-α水平显著降低（P＜0.0）,但延期灌洗组中细胞因子未见明显变化（P＞0.05）。与早期引流组相比,延期引流组的IL-1β、IL-8浓度升高,IL-10水平下降（P＜0.05）,而早期灌洗组的IL-1β、IL-8浓度降低,IL-10水平升高（P＜0.05）。早期灌洗组与延期灌洗组的腹水中IL-1β、IL-6、IL-8、IL-10和TNF-α浓度均有显著差异（P＜0.05）。结论 在SAP病程早期进行腹腔置管引流灌洗可以提高大鼠生存率。与血清中细胞因子浓度变化相比,腹水中细胞因子浓度的改变更为明显。早期引流灌洗后降低腹水中TNF-α、IL-1β、IL-6和IL-8浓度,并使IL-10浓度升高可能是提高大鼠生存率的关键因素。     

大鼠重症急性胰腺炎腹腔置管引流灌洗时机对病程转归及胰腺病理改变的影响

目的:重症急性胰腺炎(SAP)起病急遽、进展迅速、死亡率高。早期腹腔置管引流对部分病人可缓解病情。本研究拟在大鼠SAP模型中探讨腹腔引流的时机和方式,观察对病程转归和胰腺病理学形态的影响。方法:清洁级SD大鼠,随机分为SAP模型组、早期引流组、延期引流组、早期灌洗组、延期灌洗组和手术清创引流组。SAP大鼠模型采用胰胆管顺行注射牛磺胆酸钠法建立。比较各组24 h后大鼠存活率,观察胰腺病理学改变。结果:SAP大鼠模型组和清创引流组24 h后存活率差异无统计学意义。早期引流组存活率显著提高(P0.05)。采用5 mL/h生理盐水行早期灌注24 h存活率与模型组无统计学差异(P>0.05),但是采用2 mL/h生理盐水行早期灌注24 h存活率高于模型组(P     

短发卡RNA稳定沉默FOXM1对肝癌细胞体外生长的影响

目的 探讨短发夹RNA(short hairpin RNA,shRNA)表达载体稳定沉默叉头框M1(forkhead box M1,FOXM1)基因对人肝癌细胞体外生长的影响.方法 构建针对人类FOXM1mRNA的不同干扰靶点的4个shRNA表达载体,选择干扰效果最佳的表达载体和阴性对照质粒转染人肝癌细胞QGY-7703,用新霉素(G418)筛选稳定转染的克隆.用四甲基偶氮唑盐(MTT)比色法和平板克隆形成实验检测FOXM1基因沉默前后肝癌细胞体外生长能力的变化.用Annexin V-APC/PI双染法检测细胞凋亡.结果 不同人肝癌细胞株中普遍表达FOXM1蛋白.4个shRNA表达载体中,shRNA-1026表达载体的干扰效果最佳,对FOXM1 mRNA和蛋白表达水平的抑制率分别为38.5%和53.2%.用shRNA-1026稳定沉默FOXM1基因后,QGY-7703细胞增殖受到抑制,培养48、72和96 h后,沉默组的吸光值均显著低于对照组(分别t=10.830,3.578,5.734,均P＜0.05);沉默组的克隆形成能力较对照组显著降低(t=5.336,P＜0.05),而细胞凋亡较对照组显著增加(t=6.827.P＜0.05).结论 shRNA表达载体稳定沉默FOXM1基因抑制肝癌细胞的生长.

Abstract：

Objective To evaluate the effect of sustained silencing Forkhead box Ml (F0XM1) gene by short-hairpin RNA (shRNA) expression vector on cell growth of hepatocelluar carcinoma (HCC) in vitro.Methods Four shRNA expression vectors targeting different sequences of human F0XM1 mRNA were constructed.The expression vector with the best interfering effect and the negative control plasmid were used to transfect HCC cell line QGY-7703, stably transfected cell clones were selected by neomycin (G418).Cell growth was evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay and colony formation was assessed by clonogenic assay.Cell apoptosis was detected by double staining with APC conjugated Annexin V and PI.Results F0XM1 protein was detected with different levels in all these studied human cell lines.The expression vector shRNA-1026 exhibited excellent interference effect after transient transfection, which showed 38.5% and 53.2% reduction of FOXM1 mRNA and protein level respectively.The growth of QGY-7703 cells was inhibited after stable inhibition of FOXM1 expression by shRNA-1026, which was indicated by decreased absorbance value of the test group after culture for 48, 72 and 96 h compared to control group (t = 10.830,3.578 and 5.734 respectively, P ＜ 0.05).Stable inhibition of F0XM1 also led to reduced colony formation ( t = 5.336, P ＜ 0.05 ) and increased apoptosis of QGY-7703 cells in comparison to control cells (t = 6.827, P ＜ 0.05 ).Conclusions Stable silencing F0XM1 gene by shRNA suppresses the growth of HCC cells in vitro.