AIM： To examine the effects of anti-high mobility group box 1 （HIGB1） neutralizing antibody in experimental severe acute pancreatitis （SAP）. METHODS： SAP was induced by creating closed duodenal loop inC3H/HeN mi...AIM： To examine the effects of anti-high mobility group box 1 （HIGB1） neutralizing antibody in experimental severe acute pancreatitis （SAP）. METHODS： SAP was induced by creating closed duodenal loop inC3H/HeN mice. SAP was induced immediately after intrapedtoneal injection of anti-HMGB1 neutralizing antibody （200 pg）. Sevedty of pancreatitis, organ injury （liver, kidney and lung）, and bacterial translocation to pancreas was examined 12 h after induction of SAP. RESULTS： Anti-HHGB1 neutralizing antibody significantly improved the elevation of the serum amylase level and the histological alterations of pancreas and lung in SAR Anti-HHGB1 antibody also significantly ameliorated the elevations of serum alanine aminotransferase and creatinine in SAR However, anti-HHGB1 antibody worsened the bacterial translocation to pancreas. CONCLUSION： Blockade of HHGB1 attenuated the development of SAP and associated organ dysfunction, suggesting that HHGB1 may act as a key mediator for inflammatory response and organ injury in SAR展开更多
目的:探讨高迁移率族蛋白B1(high mobility group box-1,HMGB1)对宫颈癌细胞化学治疗敏感性的影响及其机制。方法:采用Western印迹法检测宫颈癌细胞HeLa和CaSki经不同药物浓度顺铂处理后LC3,Beclin1及P62的表达水平,检测使用自噬抑制剂...目的:探讨高迁移率族蛋白B1(high mobility group box-1,HMGB1)对宫颈癌细胞化学治疗敏感性的影响及其机制。方法:采用Western印迹法检测宫颈癌细胞HeLa和CaSki经不同药物浓度顺铂处理后LC3,Beclin1及P62的表达水平,检测使用自噬抑制剂和/或顺铂处理后宫颈癌细胞HeLa和CaSki中LC3,Beclin1及P62的表达水平;采用细胞计数试剂盒8(cell counting kit-8,CCK-8)检测细胞增殖水平。构建HeLa-sh HMGB1,CaSkish HMGB1,HeLa-CTR及CaSki-CTR的稳定细胞系。以CCK-8检测上述细胞系顺铂半数抑制浓度(half maximum inhibitory concentration,IC50)水平;Western印迹法检测上述细胞系中HMGB1,LC3,Beclin1及P62的表达水平。结果:在一定浓度范围内,随着顺铂药物浓度的增加,宫颈癌细胞HeLa和CaSki中LC3及Beclin1的表达增加,P62表达降低。与单用顺铂组相比,顺铂联合自噬抑制剂组细胞存活率更低(P<0.05)。在宫颈癌细胞中,HMGB1的表达与顺铂药物敏感性有关(P<0.05),与LC3,Beclin1表达呈正相关,与P62表达呈负相关。结论:HMGB1可能通过调控宫颈癌细胞内自噬的水平,影响其对顺铂的敏感性。铂类药物结合自噬抑制剂可能成为宫颈癌治疗的新策略。HMGB1可能成为预测化学治疗药物敏感性的分子标志物。展开更多
目的探讨乌司他丁对机械通气相关性肺损伤(VILI)的保护作用及相关机制。方法 24只SD大鼠,随机分为对照组、VILI组和乌司他丁治疗组(VILI+乌司他丁组)。检测肺泡灌洗液中高迁移率族蛋白1(high mobility group box-1,HMGB-1)、肿瘤坏死因...目的探讨乌司他丁对机械通气相关性肺损伤(VILI)的保护作用及相关机制。方法 24只SD大鼠,随机分为对照组、VILI组和乌司他丁治疗组(VILI+乌司他丁组)。检测肺泡灌洗液中高迁移率族蛋白1(high mobility group box-1,HMGB-1)、肿瘤坏死因子(tumor necrosis factor,TNF)-α、白细胞介素(IL)-6,肺组织中Toll样受体4(toll like receptor-4,TLR4)表达情况,肺湿干重比及肺病理损伤情况。结果 VILI组肺泡灌洗液中HMGB-1、TNF-α、IL-6水平、肺组织中TLR4表达、肺干湿重比和肺病理损伤评分均高于对照组,且差异有统计学意义(P<0.05);VILI+乌司他丁组肺泡灌洗液中HMGB-1、TNF-α、IL-6水平、肺组织中TLR4表达、肺干湿重比和肺病理损伤评分均低于VILI组,且差异有统计学意义(P<0.05)。结论乌司他丁可能通过减少HMGB-1分泌,继而减少其通过TLR4刺激肺炎症反应产生水平,达到保护VILI所致肺损伤。展开更多
高迁移率族蛋白B1(high mobility group box-1,HMGB1)是一种在哺乳动物中广泛存在、进化中高度保守的蛋白质。细胞核定位序列的存在使得生理状态下的HMGB1主要存在于细胞核内,但其在细胞内外的不同位置具有不同的生物学功能。细胞外的HM...高迁移率族蛋白B1(high mobility group box-1,HMGB1)是一种在哺乳动物中广泛存在、进化中高度保守的蛋白质。细胞核定位序列的存在使得生理状态下的HMGB1主要存在于细胞核内,但其在细胞内外的不同位置具有不同的生物学功能。细胞外的HMGB1与脓毒症、肿瘤、风湿免疫、动脉粥样硬化及缺血再灌注损伤等多种疾病密切相关。HMGB1由细胞核内迁移至细胞质再释放到细胞外的过程,涉及核定位序列的翻译后修饰、主动分泌或被动释放等机制。展开更多
OBJECTIVE In this study, RNA interference was used to evaluate the effects of HMGB1 expression on cell cycle and proliferation of the human cervical cancer cell line HeLa.METHODS We had previously constructed and scre...OBJECTIVE In this study, RNA interference was used to evaluate the effects of HMGB1 expression on cell cycle and proliferation of the human cervical cancer cell line HeLa.METHODS We had previously constructed and screened effective eukaryotic expression vectors carrying PGCsi3.0-1/ HMGB1 siRNA and PGCsi3.0-3/HMGB1 siRNA, then the vectors were transfected into HeLa cells. The expression of HMGB1 before and after transfection in HeLa cells were detected by RT-PCR and Western blot. The cell viability and proliferating activity was tested by Trypan blue dye test and MTT, and the cell cycle was determined bv flow cvtometry.RESULTS The introduction of PGCsi3.0-1/HMGB1 siRNA and PGCsi3.0-3/HMGB1 siRNA inhibited the expression of HMGB1 mRNA and protein efficiently and specifically, there was a significant difference between the siRNA groups and the control groups （P 〈 0.05）. The proliferation speed of PGCsi3.0-1 group and PGC si3.0-3 group were obviously slower than those of PGCsi3.0- Neg group and non-transfected group. Flow cytometry showed that the content of DNA in G2 phase in PGCsi3.0-1 group and PGCsi3.0-3 group were obviously more than those in PGCsi3.0- Neg group and non-transfected group, but the content in S phase was less （P 〈 0.01）. The progression of cell cycle was arrested from G2 to S phase.CONCLUSION PGCsi3.0-1/HMGB1 siRNA and PGCsi3.0-3 /HMGB1 siRNA could specially suppress the expression of HMGB1 gene, inhibit the proliferation speed of HeLa cells effectively, and arrest the progression of cell cycle from G2 to S phase. RNAi provides a new approach to the bio-therapy of cervical cancer.展开更多
AIM： To investigate the effect of high mobility group protein box-1 （HMGB1） siRNA on proliferation and apoptosis of retinoblastoma （Rb） cells.METHODS： The expression of HMGB1 in Rb cells were detected by real-ti...AIM： To investigate the effect of high mobility group protein box-1 （HMGB1） siRNA on proliferation and apoptosis of retinoblastoma （Rb） cells.METHODS： The expression of HMGB1 in Rb cells were detected by real-time polymerase chain reaction （RT-PCR） and Western blot. Chemically synthesized HMGB1 siRNA was transfected into Y79 cells. The inhibitory rate was also examined by RT-PCR and Western blot. After HMGB1 siRNA transfection, the cell proliferation was analyzed by MTT, and cell apoptosis was detected by Caspase-3 active detection kit. Cell cycle distribution and apoptosis were detected by flow cytometry. RESULTS： The expression of HMGB1 significantly elevated in Rb cells （P〈0.01）. After transfected by siRNA, the HMGB1 protein level of Y79 cells was significantly reduced （P〈0.01）. After siRNA interference HMGB1, the proportion of proliferating cells reduced, and the proportion of quiescent cells increased （P〈0.05）. In addition, apoptosis rate of Y79 cells increased from 2.03% to 9.10% after interfering with HMGB1 siRNA （P〈0.05）.CONCLUSION： Specific HMGB1 siRNA can inhibit the expression of HMGB1. The effect may be attributed to inhibit the proliferation and promote cell apoptosis.展开更多
基金Supported by Grants-in-Aid for Scientific Research from the Ministry of Education, Science, Sports and Culture of Japan and from the Ministry of Health, Labor and Welfare of Japan
文摘AIM： To examine the effects of anti-high mobility group box 1 （HIGB1） neutralizing antibody in experimental severe acute pancreatitis （SAP）. METHODS： SAP was induced by creating closed duodenal loop inC3H/HeN mice. SAP was induced immediately after intrapedtoneal injection of anti-HMGB1 neutralizing antibody （200 pg）. Sevedty of pancreatitis, organ injury （liver, kidney and lung）, and bacterial translocation to pancreas was examined 12 h after induction of SAP. RESULTS： Anti-HHGB1 neutralizing antibody significantly improved the elevation of the serum amylase level and the histological alterations of pancreas and lung in SAR Anti-HHGB1 antibody also significantly ameliorated the elevations of serum alanine aminotransferase and creatinine in SAR However, anti-HHGB1 antibody worsened the bacterial translocation to pancreas. CONCLUSION： Blockade of HHGB1 attenuated the development of SAP and associated organ dysfunction, suggesting that HHGB1 may act as a key mediator for inflammatory response and organ injury in SAR
文摘目的探讨乌司他丁对机械通气相关性肺损伤(VILI)的保护作用及相关机制。方法 24只SD大鼠,随机分为对照组、VILI组和乌司他丁治疗组(VILI+乌司他丁组)。检测肺泡灌洗液中高迁移率族蛋白1(high mobility group box-1,HMGB-1)、肿瘤坏死因子(tumor necrosis factor,TNF)-α、白细胞介素(IL)-6,肺组织中Toll样受体4(toll like receptor-4,TLR4)表达情况,肺湿干重比及肺病理损伤情况。结果 VILI组肺泡灌洗液中HMGB-1、TNF-α、IL-6水平、肺组织中TLR4表达、肺干湿重比和肺病理损伤评分均高于对照组,且差异有统计学意义(P<0.05);VILI+乌司他丁组肺泡灌洗液中HMGB-1、TNF-α、IL-6水平、肺组织中TLR4表达、肺干湿重比和肺病理损伤评分均低于VILI组,且差异有统计学意义(P<0.05)。结论乌司他丁可能通过减少HMGB-1分泌,继而减少其通过TLR4刺激肺炎症反应产生水平,达到保护VILI所致肺损伤。
文摘高迁移率族蛋白B1(high mobility group box-1,HMGB1)是一种在哺乳动物中广泛存在、进化中高度保守的蛋白质。细胞核定位序列的存在使得生理状态下的HMGB1主要存在于细胞核内,但其在细胞内外的不同位置具有不同的生物学功能。细胞外的HMGB1与脓毒症、肿瘤、风湿免疫、动脉粥样硬化及缺血再灌注损伤等多种疾病密切相关。HMGB1由细胞核内迁移至细胞质再释放到细胞外的过程,涉及核定位序列的翻译后修饰、主动分泌或被动释放等机制。
文摘OBJECTIVE In this study, RNA interference was used to evaluate the effects of HMGB1 expression on cell cycle and proliferation of the human cervical cancer cell line HeLa.METHODS We had previously constructed and screened effective eukaryotic expression vectors carrying PGCsi3.0-1/ HMGB1 siRNA and PGCsi3.0-3/HMGB1 siRNA, then the vectors were transfected into HeLa cells. The expression of HMGB1 before and after transfection in HeLa cells were detected by RT-PCR and Western blot. The cell viability and proliferating activity was tested by Trypan blue dye test and MTT, and the cell cycle was determined bv flow cvtometry.RESULTS The introduction of PGCsi3.0-1/HMGB1 siRNA and PGCsi3.0-3/HMGB1 siRNA inhibited the expression of HMGB1 mRNA and protein efficiently and specifically, there was a significant difference between the siRNA groups and the control groups （P 〈 0.05）. The proliferation speed of PGCsi3.0-1 group and PGC si3.0-3 group were obviously slower than those of PGCsi3.0- Neg group and non-transfected group. Flow cytometry showed that the content of DNA in G2 phase in PGCsi3.0-1 group and PGCsi3.0-3 group were obviously more than those in PGCsi3.0- Neg group and non-transfected group, but the content in S phase was less （P 〈 0.01）. The progression of cell cycle was arrested from G2 to S phase.CONCLUSION PGCsi3.0-1/HMGB1 siRNA and PGCsi3.0-3 /HMGB1 siRNA could specially suppress the expression of HMGB1 gene, inhibit the proliferation speed of HeLa cells effectively, and arrest the progression of cell cycle from G2 to S phase. RNAi provides a new approach to the bio-therapy of cervical cancer.
文摘AIM： To investigate the effect of high mobility group protein box-1 （HMGB1） siRNA on proliferation and apoptosis of retinoblastoma （Rb） cells.METHODS： The expression of HMGB1 in Rb cells were detected by real-time polymerase chain reaction （RT-PCR） and Western blot. Chemically synthesized HMGB1 siRNA was transfected into Y79 cells. The inhibitory rate was also examined by RT-PCR and Western blot. After HMGB1 siRNA transfection, the cell proliferation was analyzed by MTT, and cell apoptosis was detected by Caspase-3 active detection kit. Cell cycle distribution and apoptosis were detected by flow cytometry. RESULTS： The expression of HMGB1 significantly elevated in Rb cells （P〈0.01）. After transfected by siRNA, the HMGB1 protein level of Y79 cells was significantly reduced （P〈0.01）. After siRNA interference HMGB1, the proportion of proliferating cells reduced, and the proportion of quiescent cells increased （P〈0.05）. In addition, apoptosis rate of Y79 cells increased from 2.03% to 9.10% after interfering with HMGB1 siRNA （P〈0.05）.CONCLUSION： Specific HMGB1 siRNA can inhibit the expression of HMGB1. The effect may be attributed to inhibit the proliferation and promote cell apoptosis.
