目的探讨聚合人脐带血红蛋白氧载体(PolyCHb)对人乳腺癌MCF-7细胞裸鼠皮下移植瘤化疗敏感性的影响及其机制。方法收集处于指数生长期的MCF-7细胞,制成密^度为5×10^(7)个/mL的悬浮细胞,按0.2 mL/只接种于18只BALB/c-nu裸鼠右肢皮下...目的探讨聚合人脐带血红蛋白氧载体(PolyCHb)对人乳腺癌MCF-7细胞裸鼠皮下移植瘤化疗敏感性的影响及其机制。方法收集处于指数生长期的MCF-7细胞,制成密^度为5×10^(7)个/mL的悬浮细胞,按0.2 mL/只接种于18只BALB/c-nu裸鼠右肢皮下,建立裸鼠移植瘤模型,待肿瘤体积达到100 mm^(3)左右时,随机均分为化疗组:多柔比星(DOX)5 mg·kg^(-1),1次/周;化疗+PolyCHb治疗组:除给药(DOX)(同化疗组)外,PolyCHb 600 mg·kg^(-1),3次/周;对照组:生理盐水90 mg·kg^(-1),1次/周;均为经尾静脉连续注射4周。自注射当天(d0)起,每3 d 1次测量各组裸鼠肿瘤体积,据此绘制其各自(组)肿瘤生长曲线。38 d结束肿瘤生长观察,剥瘤并称取瘤重,计算抑瘤率;HE染色观察肿瘤组织病理变化,免疫组化检测肿瘤组织中HIF-1α表达,采用TUNEL法检测肿瘤细胞凋亡情况,荧光染色测定各组肿瘤组织活性氧(ROS)的含量。结果化疗+PolyCHb组、化疗组和对照组至d38时的肿瘤体积(mm^(3))分别为:196.35±103.45 vs 316.29±62.88 vs 519.42±177.33(P<0.05);化疗+PolyCHb组与化疗组抑瘤率(%)分别为62.20 vs 39.11;HE染色和TUNEL检测:化疗+PolyCHb组肿瘤组织生长区域细胞坏死和凋亡增多;免疫组化:化疗+PolyCHb组HIF-1α表达水平降低;荧光染色:化疗+PolyCHb组[活性氧(ROS)]含量增多。结论PolyCHb增加了乳腺癌裸鼠皮下移植瘤化疗的敏感性,其作用机制可能与其提高肿瘤组织内ROS含量,促进肿瘤细胞的凋亡有关。展开更多
Various mouse models to study dengue have been described by different authors, some of them using immunodeficient or some using humanized mice. Our group reported previously a deadly murine model, which used the intra...Various mouse models to study dengue have been described by different authors, some of them using immunodeficient or some using humanized mice. Our group reported previously a deadly murine model, which used the intracranial inoculum of highly virulent Dengue virus (DENV) in immune competent mouse. Here we present a model of immune competent mouse (C57BL/6), infected subcutaneously by the same highly virulent DENV (DENV3 genotype I). In this immunocompetent systemic mice model, the cytokine levels and hematological parameters such as total and differential leukocyte and platelets counts, together with weight loss, were considered important monitoring parameters, allowing a better understanding of the systemic human disease. Mice were inoculated subcutaneously and evaluated by the percentage weight variation as well as the clinical signs. Hematological parameters and cytokines levels were measured and viral titration in brain tissue or serum neutralization was performed to confirm mice infection. The subcutaneously DENV inoculated mice showed weight loss after infection, but they did not show any other clinical signs. The leukocytes and platelets decreased after subcutaneous inoculation. The cytokines TNF alpha and IFN gamma increased after infection in mice. The subcutaneous model provided scope for improved understanding of the dengue pathogenesis, as well as possible mechanism for protection to subsequent mouse infected by intracranial route in mice. This model could be used to study the vertebrate immune response and evaluation of drugs or vaccine against dengue virus.展开更多
文摘目的探讨聚合人脐带血红蛋白氧载体(PolyCHb)对人乳腺癌MCF-7细胞裸鼠皮下移植瘤化疗敏感性的影响及其机制。方法收集处于指数生长期的MCF-7细胞,制成密^度为5×10^(7)个/mL的悬浮细胞,按0.2 mL/只接种于18只BALB/c-nu裸鼠右肢皮下,建立裸鼠移植瘤模型,待肿瘤体积达到100 mm^(3)左右时,随机均分为化疗组:多柔比星(DOX)5 mg·kg^(-1),1次/周;化疗+PolyCHb治疗组:除给药(DOX)(同化疗组)外,PolyCHb 600 mg·kg^(-1),3次/周;对照组:生理盐水90 mg·kg^(-1),1次/周;均为经尾静脉连续注射4周。自注射当天(d0)起,每3 d 1次测量各组裸鼠肿瘤体积,据此绘制其各自(组)肿瘤生长曲线。38 d结束肿瘤生长观察,剥瘤并称取瘤重,计算抑瘤率;HE染色观察肿瘤组织病理变化,免疫组化检测肿瘤组织中HIF-1α表达,采用TUNEL法检测肿瘤细胞凋亡情况,荧光染色测定各组肿瘤组织活性氧(ROS)的含量。结果化疗+PolyCHb组、化疗组和对照组至d38时的肿瘤体积(mm^(3))分别为:196.35±103.45 vs 316.29±62.88 vs 519.42±177.33(P<0.05);化疗+PolyCHb组与化疗组抑瘤率(%)分别为62.20 vs 39.11;HE染色和TUNEL检测:化疗+PolyCHb组肿瘤组织生长区域细胞坏死和凋亡增多;免疫组化:化疗+PolyCHb组HIF-1α表达水平降低;荧光染色:化疗+PolyCHb组[活性氧(ROS)]含量增多。结论PolyCHb增加了乳腺癌裸鼠皮下移植瘤化疗的敏感性,其作用机制可能与其提高肿瘤组织内ROS含量,促进肿瘤细胞的凋亡有关。
文摘Various mouse models to study dengue have been described by different authors, some of them using immunodeficient or some using humanized mice. Our group reported previously a deadly murine model, which used the intracranial inoculum of highly virulent Dengue virus (DENV) in immune competent mouse. Here we present a model of immune competent mouse (C57BL/6), infected subcutaneously by the same highly virulent DENV (DENV3 genotype I). In this immunocompetent systemic mice model, the cytokine levels and hematological parameters such as total and differential leukocyte and platelets counts, together with weight loss, were considered important monitoring parameters, allowing a better understanding of the systemic human disease. Mice were inoculated subcutaneously and evaluated by the percentage weight variation as well as the clinical signs. Hematological parameters and cytokines levels were measured and viral titration in brain tissue or serum neutralization was performed to confirm mice infection. The subcutaneously DENV inoculated mice showed weight loss after infection, but they did not show any other clinical signs. The leukocytes and platelets decreased after subcutaneous inoculation. The cytokines TNF alpha and IFN gamma increased after infection in mice. The subcutaneous model provided scope for improved understanding of the dengue pathogenesis, as well as possible mechanism for protection to subsequent mouse infected by intracranial route in mice. This model could be used to study the vertebrate immune response and evaluation of drugs or vaccine against dengue virus.
