大黄灵仙胶囊调控小鼠胆囊结石形成机制中BSEP、NTCP基因表达的研究

Dahuang Lingxian Capsule in regulation of BSEP and NTCP gene expressions in the gallstone formation mechanism of mice

目的研究大黄灵仙胶囊调控小鼠肝胆管侧膜细胞上胆盐输出泵(BSEP)、钠离子-牛磺胆酸共转运蛋白(NTCP)的蛋白及mRNA表达水平,干预胆囊结石形成的作用机制。方法将60只SPF级C57BL/6雄性小鼠随机分为普食组(P组)、致石组(Z组)、致石+灌水组(Z+S组)、致石+大黄灵仙胶囊组(Z+D组)、致石+熊去氧胆酸组(Z+X组),每组各12只,喂养8周,1天/次。采取小鼠胆囊组织观察造模情况,运用Western Blotting及RT-PCR技术对肝脏组织中BSEP、NTCP的蛋白及mRNA表达量进行分析并对比其差异。结果Z组结石形成数多于P组,差异有统计学意义(P<0.001)。Western Blotting:Z+D组和Z+X组BSEP、NTCP蛋白表达水平无明显差异(P>0.05),Z+D组高于Z+S组,差异具有统计学意义(P<0.05)。RT-PCR:Z+S组、Z+D组与Z+X组BSEP、NTCP mRNA的转录水平存在差异,Z+S组<Z+D组、Z+S组<Z+X组,差异有统计学意义(P<0.05)。结论致石性饲料喂养可致小鼠胆囊结石形成,大黄灵仙胶囊可以增强小鼠肝胆管侧膜细胞上BSEP、NTCP的蛋白及mRNA表达水平干预胆囊结石形成。

Objective To study the mechanism of how Dahuang Lingxian Capsules regulate the protein and mRNA expressions of bile salt export pump(BSEP)and Na+-taurocholate cotransporting polypeptide(NTCP)in the lateral membrane cells of the hepatic bile ducts of mice for intervening the formation of gallstone.Methods Sixty healthy male SPF C57BL/6 mice were randomly divided into 5 groups:general feeding group(group P),lithogenic diet group(group Z),lithogenic diet+water injecting group(Z+S group),lithogenic diet+Dahuang Lingxian Capsule injecting group(Z+D group),lithogenic diet+ursodeoxycholic acid injecting group(Z+X group),with 12 mice in each group,fed once daily for 8 weeks.The gallbladder tissue of mice was collected to observe the modeling.Western blotting and real-time PCR were used to detect and analyze the protein and mRNA expressions of BSEP,NTCP in liver tissue,which would also be compared.Results The stones formed in group Z were more than those in group P and the difference was statistically significant(P<0.05).Western Blotting showed that there was no significant difference in protein expressions of BSEP and NTCP between Z+D group and Z+X group(P>0.05).The protein expressions of BSEP and NTCP in Z+D group were higher than those in the Z+S group and the difference was statistically significant(P<0.05).RT-PCR showed that the mRNA transcription levels of BSEP and NTCP in Z+S group,Z+D group and Z+X group were significantly different.The mRNA levels of the Z+S group were lower than those of the Z+D group,while those of the Z+S group were lower than those of the Z+X group,which showed statistically significant differences(P<0.05).Conclusion Lithogenic feeding can cause gallstone formation in mice.Dahuang Lingxian Capsules can enhance the protein and mRNA expressions of BSEP and NTCP in the lateral membrane cells of hepatic bile ducts to intervene the formation of gallstone in mice.