猪苓多糖对四氯化碳诱导的建鲤肝细胞损伤中生化指标及CYP3A表达的影响

Effects of polyporus polysaccharides on biochemical indexes and CYP3A expression of carbon tetrachloride injured primary hepatocytes of Jiancarp

采用胰酶消化法分离原代建鲤肝细胞并进行培养,用四氯化碳(CCl4)构建建鲤肝细胞损伤模型,以3种不同浓度的猪苓多糖进行干预,检测肝细胞培养液中丙氨酸转氨酶(GPT)、天冬氨酸转氨酶(GOT)、乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)、丙二醛(MDA)的含量及肝细胞存活率;收集肝细胞进行RNA提取,并用RT-PCR法测定CYP3A基因表达情况。结果表明:以前处理组中质量浓度为0.8mg/mL的猪苓多糖效果最好,与CCl4组相比,显著降低了GOT、GPT、MDA在肝细胞中的释放;极显著降低了LDH在肝细胞中的释放;显著升高了肝细胞中SOD活性值;显著提高了肝细胞的存活率;显著诱导了CYP3A mRNA的表达。这说明猪苓多糖能有效抑制CCl4所造成的建鲤肝细胞损伤。

In order to study the protect effects of polyporus polysaccharide on hepatocytes injured by carbon tetrachloride（CCl4）,the primary hepatocytes in Jian carp were isolated by trypsin digestion method,cultured in vitro and induced by CCl4.The hepatocytes were treated with polyporus polysaccharide of different concentrations,the culture medium of hepatocytes were then collected,the content of alanine aminotransferase（GPT）,aspartate aminotransferase（GOT）,superoxide dismutase（SOD）,malondialdehyde（MDA）,lactic acid dehydrogenase（LDH）,and cell livability of primary cultured hepatocytes were determinated.Total RNA in primary cultured hepatocytes was extracted with trizol reagent and the mRNA levels of CYP3A were determined by RT-PCR.The results showed that in the pre-treatment group at 0.8mg/mL of polyporus polysaccharide,the release of GOT,GPT,MDA were significantly decreased,the activity of SOD was increased,the release of LDH was decreased,the cell livability and the expression of CYP3Aat mRNA level were increased,indicating best protect effect of this group.It can be concluded from the data obtained that polyporus polysaccharides could effectively protect the primary cultured hepatocytes against CCl4induced injury.