紫杉醇脂质复合体对大鼠肝移植后血管病变的抑制作用
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摘要
目的:免疫反应是导致移植物血管病变(allograft vasculopathy AV)的核心因素,急性排斥反应所致移植物血管损伤常常是细胞免疫和体液免疫共同致病的结果,其作用机理相当复杂,治疗比较棘手。此项研究在于通过纳米生物技术探讨紫杉醇脂质复合体在大鼠肝移植动物模型中治疗急性排斥反应所致移植肝血管损伤的价值。
方法:本研究内容分为二大部分:第一部分为紫杉醇脂质复合体的制备,并对紫杉醇脂质复合体药代动力学及生物安全性进行检测。第二部分通过建立大鼠肝移植模型,利用酶联免疫吸附试验(ELISA),分三组(紫杉醇脂质复合体组、紫杉醇组和环孢素组)检测大鼠肝移植术后2周血清血管内皮生长因子(VEGF)、肿瘤坏死因子-α(TNF-α)、白介素-2(IL-2)含量及大鼠移植肝脏组织标本中细胞周期素依赖性激酶2(CDK2)和细胞周期素蛋白E(Cyclin E)的表达,利用末端脱氧核糖核苷酸转移酶介导的缺口末端标记法(TUNEL)检测血管平滑肌细胞(VSMC)的凋亡率,探讨紫杉醇纳米脂质体抑制大鼠肝移植急性排斥反应所致血管损伤的机制。对紫杉醇脂质复合体在抑制急性排异所致移植肝血管病变方面作出评价。
结果:紫杉醇脂质复合体的平均粒径为115.1nm,包封率为65.3%。具有明显的肝靶向性、良好的细胞和组织相容性。紫杉醇脂质复合体能显著降低肝血管平滑肌细胞(VSMC)中血管细胞周期素依赖性激酶2(CDK2)和细胞周期素蛋白E(Cyclin E)的表达,使血管平滑肌细胞(VSMC)凋亡明显增加,抑制VSMC增殖;并能减少大鼠肝移植后免疫反应过程中肿瘤坏死因子-α(TNF-α)、白介素-2(IL-2)、血管内皮生长因子(VEGF)的产生,其效果明显优于注射纯紫杉醇和环孢素组。
结论:紫杉醇脂质复合体具有明显的肝靶向性,能减轻大鼠肝移植后急性排斥反应所致血管的损伤,紫杉醇脂质复合体作为一种新型辅助免疫抑制剂在减轻大鼠肝移植后急性血管损害、减轻免疫排斥反应方面有潜在应用价值。
Purpose:Immunoreaction trigger the main injuries of allograft vasculopathy.Cellular immunity and humoral immunity cause the injuries of allograft vasculopathy in acute rejection.The mechanisn of this injuries is very complex.The effect of routine antirejection therapy is troublesome. This experiment is to explore the value of paclitaxel-liposome compound and its effect of therapy on allograft vasculopathy in acute rejection after liver transplantion in rats by using nano-biotechnology.
Methods:The contents of this paper include two parts.The first part is the preparaty of paclitaxel-liposome compound and its pharmacokinetics and biologic security.The second part is to describe the proess and methods in rats liver tranplantion models.Sprague-Dawley rats were randomly divided into three groups(CsA group、paclitaxel group、paclitaxel-liposome compound group).Two week late,enzyme linked immunosorbent assay(ELISA)were performed for testing the levels of serum VEGF、TNF-a、IL-2 and the expressions of CDK2、Cyclin E protein.VSMC apoptosis in hepatic tissues was evaluated by using the technique of terminal deoxynucleotidyl transferasebiotin nick end-labeling(TUNEL). The purpose of this paper is to investigate inhibitory effect of the paclitaxel-liposome compound on vasculopathy damage in acute rejection after liver transplation in rats.
Result:The result showed the mean diameter and drug envelopment rate of paclitaxel-liposome compound were 115.1nm,65.3%respectively.The paclitaxel-liposome compound targeted evidently to liver and was very biocompatible.The paclitaxel-liposome compound could significantly down-regulate the express of CDK2 and Cyclin E protein,and increase VSMC apoptosis,also inhibite the vasularization,also could reduce the produce of VEGF、TNF-a、IL-2.The inhibitory effect of the paclitaxel-liposome compound group on allograft vasculopathy significantlly surpassed that of paclitaxel group and CsA group.
Conclution:The paclitaxel-liposome compound argeted evidently to liver and was very biocompatible.It could lighten the injuiris of allograft vasculopathy in acute rejection after liver transplation in rats.As a new aided immunity inhibitory preparation.The paclitaxel-liposome compound has its potention value for lessening the allograft vasculopathy injuiris in acute rejection after liver transplation.
方法:本研究内容分为二大部分:第一部分为紫杉醇脂质复合体的制备,并对紫杉醇脂质复合体药代动力学及生物安全性进行检测。第二部分通过建立大鼠肝移植模型,利用酶联免疫吸附试验(ELISA),分三组(紫杉醇脂质复合体组、紫杉醇组和环孢素组)检测大鼠肝移植术后2周血清血管内皮生长因子(VEGF)、肿瘤坏死因子-α(TNF-α)、白介素-2(IL-2)含量及大鼠移植肝脏组织标本中细胞周期素依赖性激酶2(CDK2)和细胞周期素蛋白E(Cyclin E)的表达,利用末端脱氧核糖核苷酸转移酶介导的缺口末端标记法(TUNEL)检测血管平滑肌细胞(VSMC)的凋亡率,探讨紫杉醇纳米脂质体抑制大鼠肝移植急性排斥反应所致血管损伤的机制。对紫杉醇脂质复合体在抑制急性排异所致移植肝血管病变方面作出评价。
结果:紫杉醇脂质复合体的平均粒径为115.1nm,包封率为65.3%。具有明显的肝靶向性、良好的细胞和组织相容性。紫杉醇脂质复合体能显著降低肝血管平滑肌细胞(VSMC)中血管细胞周期素依赖性激酶2(CDK2)和细胞周期素蛋白E(Cyclin E)的表达,使血管平滑肌细胞(VSMC)凋亡明显增加,抑制VSMC增殖;并能减少大鼠肝移植后免疫反应过程中肿瘤坏死因子-α(TNF-α)、白介素-2(IL-2)、血管内皮生长因子(VEGF)的产生,其效果明显优于注射纯紫杉醇和环孢素组。
结论:紫杉醇脂质复合体具有明显的肝靶向性,能减轻大鼠肝移植后急性排斥反应所致血管的损伤,紫杉醇脂质复合体作为一种新型辅助免疫抑制剂在减轻大鼠肝移植后急性血管损害、减轻免疫排斥反应方面有潜在应用价值。
Purpose:Immunoreaction trigger the main injuries of allograft vasculopathy.Cellular immunity and humoral immunity cause the injuries of allograft vasculopathy in acute rejection.The mechanisn of this injuries is very complex.The effect of routine antirejection therapy is troublesome. This experiment is to explore the value of paclitaxel-liposome compound and its effect of therapy on allograft vasculopathy in acute rejection after liver transplantion in rats by using nano-biotechnology.
Methods:The contents of this paper include two parts.The first part is the preparaty of paclitaxel-liposome compound and its pharmacokinetics and biologic security.The second part is to describe the proess and methods in rats liver tranplantion models.Sprague-Dawley rats were randomly divided into three groups(CsA group、paclitaxel group、paclitaxel-liposome compound group).Two week late,enzyme linked immunosorbent assay(ELISA)were performed for testing the levels of serum VEGF、TNF-a、IL-2 and the expressions of CDK2、Cyclin E protein.VSMC apoptosis in hepatic tissues was evaluated by using the technique of terminal deoxynucleotidyl transferasebiotin nick end-labeling(TUNEL). The purpose of this paper is to investigate inhibitory effect of the paclitaxel-liposome compound on vasculopathy damage in acute rejection after liver transplation in rats.
Result:The result showed the mean diameter and drug envelopment rate of paclitaxel-liposome compound were 115.1nm,65.3%respectively.The paclitaxel-liposome compound targeted evidently to liver and was very biocompatible.The paclitaxel-liposome compound could significantly down-regulate the express of CDK2 and Cyclin E protein,and increase VSMC apoptosis,also inhibite the vasularization,also could reduce the produce of VEGF、TNF-a、IL-2.The inhibitory effect of the paclitaxel-liposome compound group on allograft vasculopathy significantlly surpassed that of paclitaxel group and CsA group.
Conclution:The paclitaxel-liposome compound argeted evidently to liver and was very biocompatible.It could lighten the injuiris of allograft vasculopathy in acute rejection after liver transplation in rats.As a new aided immunity inhibitory preparation.The paclitaxel-liposome compound has its potention value for lessening the allograft vasculopathy injuiris in acute rejection after liver transplation.
引文
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