Gu J J, Yao M, Cai Y, Fang M, Wang L, Zheng W J, Yao D B, Dong Z Z, Yao D F
Medical School of Nantong University, Jiangsu 226001, China.
Affiliated Hospital of Nantong University, Jiangsu 226001, China.
Zhonghua Gan Zang Bing Za Zhi. 2017 Apr 20;25(4):279-284. doi: 10.3760/cma.j.issn.1007-3418.2017.04.009.
To investigate the dynamic expression of hepatic carnitine palmitoyltransferase-II (CPT-II) in the mitochondrial inner membrane during hepatocyte malignant transformation induced by lipid accumulation. Male Sprague-Dawley rats were divided randomly into control, fatty liver, and induced cancer groups, which were fed with normal, high-fat (HF), and HF containing 2-fluorenylacetamide (0.05%, 2-FAA) diets, respectively, for 14 weeks. One rat from each group was sacrificed every two weeks and the blood and liver samples were collected. Liver morphological changes were evaluated with hematoxylin and eosin staining, and the liver tissue samples were divided into control, fatty liver, degeneration, precancerous, and cancerous groups accordingly. Hepatic lipids were dyed by the oil red O method. The CPT-II expression was measured by immunohistochemistry and compared with the specific CPT-II concentration (ng/mg liver protein, ng/mg P) among different groups. Serum levels of circulating total cholesterol (Tch), triglyceride (TG), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) were quantitatively analyzed. Massive lipid accumulation hepatocytes was seen in rats on HF and HF containing 2-FAA diets. The lipid levels in the control group were significantly lower than those in the fatty liver ( = -11.556, < 0.001), degeneration ( = -4.847, = 0.04), precancerous ( = -13.652, = 0.005), and cancerous groups ( = -10.896, = 0.008). The serum TG and Tch levels in the degeneration, precancerous, and cancerous groups were 2-3 times higher than those in the control group ( < 0.05). After 2-FAA treatment, the morphological changes of rat hepatocytes showed the progression from degeneration and precancerosis to cancerosis, with hepatocyte injury. The serum AST and ALT levels in the degeneration, precancerous, and cancerous groups were significantly higher (4-8 times) than those in the control group ( < 0.05). The specific concentration of liver CPT-II expression was significantly reduced during hepatocyte malignant transformation, as confirmed by immunohistochemistry, with the CPT-II levels significantly lower in the cancerous group than in any of other groups ( < 0.05). Low hepatic CPT-II expression might lead to abnormal lipid accumulation in hepatocytes, which should promote the malignant transformation of hepatocytes.
为研究脂质蓄积诱导肝细胞恶性转化过程中线粒体内膜上肝肉碱棕榈酰转移酶-II(CPT-II)的动态表达。将雄性Sprague-Dawley大鼠随机分为对照组、脂肪肝组和诱癌组,分别给予正常饮食、高脂(HF)饮食以及含2-芴基乙酰胺(0.05%,2-FAA)的高脂饮食,持续14周。每隔两周处死每组中的1只大鼠并采集血液和肝脏样本。用苏木精-伊红染色评估肝脏形态变化,并据此将肝组织样本分为对照组、脂肪肝组、变性组、癌前组和癌组。采用油红O法对肝脏脂质进行染色。通过免疫组织化学检测CPT-II表达,并比较不同组间CPT-II的特异性浓度(ng/mg肝脏蛋白,ng/mg P)。对血清中总胆固醇(Tch)、甘油三酯(TG)、丙氨酸氨基转移酶(ALT)和天冬氨酸氨基转移酶(AST)水平进行定量分析。在给予HF饮食和含2-FAA的HF饮食的大鼠中可见大量脂质蓄积的肝细胞。对照组的脂质水平显著低于脂肪肝组(t = -11.556,P < 0.001)、变性组(t = -4.847,P = 0.04)、癌前组(t = -13.652,P = 0.005)和癌组(t = -10.896,P = 0.008)。变性组、癌前组和癌组的血清TG和Tch水平比对照组高2 - 3倍(P < 0.05)。经2-FAA处理后,大鼠肝细胞的形态变化显示从变性和癌前病变发展为癌变,并伴有肝细胞损伤。变性组、癌前组和癌组的血清AST和ALT水平显著高于对照组(4 - 8倍)(P < 0.05)。免疫组织化学证实,在肝细胞恶性转化过程中肝脏CPT-II表达的特异性浓度显著降低,癌组的CPT-II水平显著低于其他任何组(P < 0.05)。肝脏CPT-II低表达可能导致肝细胞脂质蓄积异常,进而促进肝细胞恶性转化。