Kim Ki-Young, Cho Syung-Eun, Yu Byung-Soo
Dept. of Pathology, Medical School and Professional Graduate School of Oriental Medicine, Korea.
22Bionanochemistry Division, Natural Science College, Wonkwang University, 344-2 IKsan City, Jeonbook, Korea.
Toxicol Res. 2009 Jun;25(2):71-78. doi: 10.5487/TR.2009.25.2.071. Epub 2009 Jun 1.
We investigated the pathogenesis of liver tissue damage during the lipid peroxidation and fibrogenesis with the observation of correlations between the parameters of collagen synthesis (and deposition) and lipid peroxidation in liver fibrosis (cirrhosis) rats. Rats were randomly divided into two groups, normal and CCl-thiopental sod. intoxicated group. And the one group was treated intragastrically with the mixture of CCl-thiopental sod. 3 times per week for 3 weeks. The liver tissue and sera were used for the measurement of hydroxyproline (HYP), malonedialdehyde (MDA) and superoxide dismutase (SOD). Biochemical parameters such as aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), total-bilirubin and blood urea nitrogen (BUN) were measured. Additionally, the expression of collagen α1(III) and β-actin mRNA was observed by RT-PCR. The histological change in liver tissue was also observed by Masson's trichrome and H&E staining. Correlation analysis was carried by Spearman's rho method. All biochemical parameters except total-bilirubin were significantly higher in the CCl-thiopental sod. treated group than that of the normal group ( < 0.01). In the CCl-thiopental sod. treated group, Hyp as a parameter of collagen synthesis (deposition) and MDA as a metabolite of lipid peroxidation, were significantly elevated by 1.98 and 2.11 times higher than that of the normal group (> 0.001) respectively. The activity of SOD in the CCl-thiopental sod. treated group is decreased significantly by 44.8% (> 0.001). And collagen α1(III) mRNA was more expressed in the CCl-thiopental sod. treated group than that of the normal group. However, the expression of β-actin mRNA is showed similar in both of groups. A good correlation was observed between the content of hyp and MDA concentration (r = 0.70, n = 40) in the two groups. And the correlation between the levels of hyp and SOD (r = -0.71, n = 25) is also reliable. However, no correlation were observed between MDA concentration and SOD (r = -0.40, n = 25) in the two groups. Elevated levels of MDA in CCl-thiopental sod. treated rats indicated enhancement of lipid peroxidation, which is accompanied by a decrease in SOD activity. Moreover, we could confirm that the parameters of collagen synthesis (and deposition) is in good correlation with the metabolite of lipid peroxidation (MDA) and the lipid peroxidation antagonizing enzyme (SOD). Hence, we propose that lipid peroxidation and collagen synthesis (and deposition) could be enhanced by intragastrically application of CCl-thiopental sod. during a short terms. And ➁ the intoxication of CCl-thiopental sod. could be used for monitoring of lipid peroxidation and collagen synthesis (and depositon) for test of antioxidant and antifibrotic agent.
我们通过观察肝纤维化（肝硬化）大鼠胶原合成（及沉积）参数与脂质过氧化之间的相关性，研究脂质过氧化和纤维化形成过程中肝组织损伤的发病机制。大鼠随机分为两组，正常组和四氯化碳 - 硫喷妥钠中毒组。其中一组每周经胃内给予四氯化碳 - 硫喷妥钠混合物3次，共3周。采集肝组织和血清用于测定羟脯氨酸（HYP）、丙二醛（MDA）和超氧化物歧化酶（SOD）。检测生化参数如天冬氨酸转氨酶（AST）、丙氨酸转氨酶（ALT）、碱性磷酸酶（ALP）、总胆红素和血尿素氮（BUN）。此外，通过RT-PCR观察胶原α1(III)和β-肌动蛋白mRNA的表达。采用Masson三色染色和苏木精 - 伊红染色观察肝组织的组织学变化。采用Spearman等级相关法进行相关性分析。除总胆红素外，四氯化碳 - 硫喷妥钠处理组的所有生化参数均显著高于正常组（<0.01）。在四氯化碳 - 硫喷妥钠处理组中，作为胶原合成（沉积）参数的Hyp和作为脂质过氧化代谢产物的MDA分别比正常组显著升高1.98倍和2.11倍（>0.001）。四氯化碳 - 硫喷妥钠处理组的SOD活性显著降低44.8%（>0.001）。并且胶原α1(III)mRNA在四氯化碳 - 硫喷妥钠处理组中的表达高于正常组。然而，β-肌动蛋白mRNA在两组中的表达相似。两组中Hyp含量与MDA浓度之间存在良好的相关性（r = 0.70，n = 40）。Hyp水平与SOD之间的相关性（r = -0.71，n = 25）也可靠。然而，两组中MDA浓度与SOD之间未观察到相关性（r = -0.40，n = 25）。四氯化碳 - 硫喷妥钠处理的大鼠中MDA水平升高表明脂质过氧化增强，同时伴有SOD活性降低。此外，我们可以确认胶原合成（及沉积）参数与脂质过氧化代谢产物（MDA）和脂质过氧化拮抗酶（SOD）具有良好的相关性。因此，我们提出，短期内经胃内给予四氯化碳 - 硫喷妥钠可增强脂质过氧化和胶原合成（及沉积）。并且，四氯化碳 - 硫喷妥钠中毒可用于监测脂质过氧化和胶原合成（及沉积），以测试抗氧化剂和抗纤维化药物。
