粉防己碱在硫代乙酰胺和胆道接扎诱导大鼠肝纤维化中的作用

论文摘要

目的:观察粉防已碱诱导的肝星状细胞凋亡以及检测粉防已碱在硫代乙酰胺和胆道结扎模型中抗肝纤维化的作用。 方法:在体外实验中,我们观察了对T-HSC/C1-6的凋亡作用;在体内实验中,我们建立了硫代乙酰胺和胆道结扎两种肝纤维化模型,粉防已碱口服给药5,10,20mg/kg剂量,阳性对照组采用γ-干扰素进行对照,给药28天后,采血测量血清学丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）、总胆红素（T-Bil）、羟脯氨酸（Hyp）、透明质酸（HA）和层粘连蛋白（LN）指标的变化,同时观察组织病理学的变化。结果:在体外实验研究中,显示T-HSC/C1-6上处理0.5-25μM粉防已碱后随着浓度的增加生存力逐渐降低。在体内实验中,粉防已碱和γ-干扰素可以明显降低硫代乙酰胺和胆道结扎导致的血清学指数的升高,其组织病理切片反映粉防已碱可以改善肝纤维化程度,其中高剂量组20mg/kg与γ-干扰素组比较效果更佳,但无统计学意义。 结论:体外试验表明粉防已碱可诱导肝星状细胞凋亡;体内试验表明粉防已碱能够改善硫代乙酰胺和胆道结扎导致的肝纤维化程度。

论文目录

• Abstract
• Abstract （Chinese）
• Abstract （English）
• Introduction
• Chapter 1. Induction of Apoptosis by tetrandrine in activated hepatic stellate cells
• 1. Reagents and Apparatus
• 2. Methods
• 2.1 Isolation of hepatic stellate cells and establishment of T-HSC/C1-6 cell line
• 2.2 Cell viability assay
• 2.3 Morphology observation
• 2.4 DNA fragmentation analysis
• 2.5 Statistical analysis
• 3. Results
• 3.1 Effect of tetrandrine on cell viability
• 3.2 Effect of tetrandrine on morphology and DNA fragmentation of T-HSC/C1-6
• 4. Discussion
• Chapter 2. Antifibrotic effect of tetrandrine on hepatic fibrosis induced by thioacetamide
• 1 Materials
• 1.1 plant material
• 1.2 Reagents and Apparatus
• 1.3 Animals
• 2. Methods
• 2.1 Experimental model design
• 2.2 Determination of serum biochemical parameters
• 2.3 Determination of hydroxyproline contents in liver
• 2.4 Determination of HA and LN contents in liver
• 2.5 Histology and immunohistochemical examination of liver
• 2.5.1 Morphological studies
• 2.5.2 Sirius red staining
• 2.5.3 a-SMA
• 2.6 Statistical analysis
• 3. Results
• 3.1 Liver/weight index
• 3.2 The effect of the tetrandrine suppressed rats serum biochemical parameters After TAA administration
• 3.3 The effect of the tetrandrine suppressed rats hydroxyproline matrix contents after TAA administration
• 3.4 Extracellular matrix content of HA and LN in liver
• 3.5 The effects of tetrandrine on histological changes in the rats liver
• 4. Discussion
• Chapter 3. Antifibrotic effect of tetrandrine in fibrotic induced by Bile Duct Ligation and Scission in Rats
• 1. Materials
• 1.1 plant material
• 1.2 Reagents and Apparatus
• 1.3 Animals
• 2. Methods
• 2.1 Experimental model design
• 2.2 Determination of serum biochemical parameters
• 2.3 Determination of hydroxyproline contents in liver
• 2.4 Determination of HA and LN contents in liver
• 2.5 Light microscopy
• 2.6 Statistical analysis
• 3. Results
• 3.1 Liver/weight index
• 3.2 The effect of the tetrandrine suppressed rats serum biochemical parameters after Bile duct ligation
• 3.3 The effect of the tetrandrine suppressed rats hydroxyproline matrix contents after BDL administration
• 3.4 Extracellular matrix content of HA and LN in liver
• 3.5 The effects of tetrandrine on histological changes in the rats liver induced by BDL
• 4. Discussion
• 5. Conclusion
• References
• Review
• Acknowledgment