摘要
目的：目前认为肝纤维化尚存在可逆性。实验以光镜和细胞图像胶原半定量手段进一步验证大豆磷脂干预复合病因诱导肝纤维化大鼠模型肝细胞的组织学变化特点。方法：实验于2004-10/2007-05在辽宁医学院科学实验中心完成。①实验材料及分组：健康雄性SD大鼠24只，体质量180～220g。随机分为3组，即对照组、模型组、大豆磷脂组，每组8只。②实验过程：模型组采用复合病因诱导大鼠肝纤维化（给预高脂低蛋白食物和饮乙醇，皮下注射四氯化碳），大豆磷脂组给予造模饮食水，同时给予大豆磷脂300mg/（kg·d）灌胃，正常对照组给予大鼠正常饮食水。42d后麻醉下断颈处死大鼠。③实验评估：验证模型是否成功：采用光镜观察肝组织病理学改变；并用细胞图像分析仪对肝纤维化进行半定量分析；检测各组血清谷丙转氨酶、谷草转氨酶、单胺氧化酶活性。实验过程对动物的处置符合动物伦理学标准。结果：①造模结果：6周末验证大鼠肝纤维化模型造模成功。②光镜观察组织学变化：苏木精-伊红染色可见模型组正常肝小叶已经被破坏，坏死区中央或周围有炎性细胞浸润。肝窦多受压变窄或闭塞。门脉区有大量成纤维细胞、纤维细胞和增生的胶原纤维，纤维组织相互环绕形成大量假小叶。MASSON染色可见模型组肝组织形成了较宽的以胶原纤维为主要成分的纤维间隔（绿色），胶原纤维分隔、包绕肝小叶，多数形成假小叶。大豆磷脂组上述变化较轻。③细胞图像胶原半定量测定结果：模型组、大豆磷脂组肝组织胶原纤维面积密度显著高于正常对照组（P〈0．01），而大豆磷脂组的指标显著低于模型组（P〈0．01）。④血清谷丙转氨酶、谷草转氨酶及单胺氧化酶活性：模型组和大豆磷脂组高于正常对照组（P〈0．01），但大豆磷脂组的指标低于模型组（P〈0．01）。结论：大豆磷脂干预肝纤维化大鼠后肝模型血清酶活性比模型组明显下降，胶原积累程度半定量分析也验证了干预后胶原纤维密度低于模型组，说明干预修复了受损的肝细胞。
AIM： It is believed that liver fibrosis is reversible. With light microscope and a semiquantitative collagen analysis using imaging, this experiment observed the preventive effects of soybean phospholipid on histological changes of liver cells in multiple factors-induced liver fibrosis in rats.
METHODS： Experiments were performed at the Center of Scientific Experiments of Liaoning Medical University from October 2004 to May 2007. There were 24 healthy male rats ranged 180-220 g in weight. Rats were randomly divided into three groups： control （n=8）, model （n=8） and soybean phospholipid （n=8） groups. Multiple factors-induced liver fibrosis models were established in rats from the model group （receiving high-lipid low-protein food and alcohol, carbon tetrachloride via subcutaneous injection）. Rats in soybean phospholipid group were treated with soybean phospholipid （300 mg/kg per day） and water. Rats of the normal control group were treated with water. After 42 days of multiple factors, rats were sacrificed. After model verification, the pathology change of hepatic tissue was observed by light microscope; Liver fibrosis was analyzed with image analytic system by semiquantitative method. The activities of serum glutamic pyruvic transaminase, aspartate transaminase and monoamine oxidase were measured. Animal intervention in the experiment met the animal ethical standards. RESULTS： Rat models of liver fibrosis were successfully established 6 weeks later. Histological changes by light microscope： Hematoxylin-eosin staining showed that hepatic lobules were damaged in the model group, and inflammatory cell infiltration was present in the central or surrounding zone of necrosis. Sinus hepaticus was compressed or occluded. A mass of fibroblasts, fibrocytes and proliferative collagenous fibers were seen in portal vein. Many false folioles surrounded fibrous tissues. MASSON staining showed that multiple collagen fibrillar septation and pseud-lobulus was formed in hepatic tissue of model group. The extent of liver damages was less in the soybean phospholipid group than in the model group. Semiquantitative collagen analysis using imaging： A area of density of collagen fibers in hepatic tissue was significantly more in the model and soybean phospholipid groupe than in the normal control group （P 〈 0.01）, whereas it was significantly lower in the soybean phospholipid group than in the model group （P 〈 0.01）. The activities of serum glutamic pyruvic transaminase, aspartate transaminase and monoamine oxidase were higher in the model and soybean phospholipid groups than in the normal control group （P 〈 0.01）, whereas it was significantly lower in the soybean phospholipid group than in the model group （P 〈 0.01）.
CONCLUSION： Activity of serum enzyme is significantly reduced in liver fibrosis rats after intervening with soybean phospholipid compared to the model group. Semiquantitative collagen analysis using imaging verified that the density of collagenous fibers was lower after intervention than the model group. It is concluded that the intervention repairs the damaged liver cells.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2008年第20期3914-3917,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
