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基于免疫应激模型,研究何首乌经“九蒸九晒”炮制后对特异质肝损伤大鼠的代谢作用机制。将何首乌和制首乌的50%乙醇提取物单独灌胃或联合无毒剂量的脂多糖(LPS)给予SD大鼠,检测血清丙氨酸氨基转氨酶(ALT)、天冬氨酸氨基转氨酶(AST),HE染色观察肝脏病理学改变,对比考察何首乌炮制前后单用或联合LPS的毒性差异。通过UPLC-QTOF/MS分析不同组别大鼠血浆样品的代谢轮廓谱,结合偏最小二乘判别法(OPLS-DA)的分析方法,研究何首乌经“九蒸九晒”炮制前后对特异质肝损伤大鼠血浆中内源性代谢物的影响,寻找何首乌炮制减毒的潜在生物标志物,将鉴定到的生物标志物输入KEGG数据库中构建代谢通路。结果显示,何首乌炮制前后单次灌胃(5.4 g·kg~(-1))对大鼠ALT和AST无显著影响,肝脏切片未见明显病理学改变;而相同剂量的何首乌和制首乌联合LPS给药后,何首乌组ALT和AST均显著升高(P<0.01),肝脏切片可见明显病理学改变,而制首乌组未出现肝损伤。主成分分析(PCA)结果显示,正常对照组、LPS组、LPS/制首乌组和LPS/何首乌组血清代谢物谱得到明显分离,发现并鉴定了10个与肝损伤相关的潜在生物标志物。推测这些生物标志物可能与鞘脂代谢、亚油酸代谢、甾类激素生物合成、半乳糖代谢、类固醇生物合成、细胞色素P450外源性物质代谢、嘧啶代谢、不饱和脂肪酸合成、初级胆汁酸合成,以及牛磺酸与亚牛磺酸代谢等10个代谢通路有关。故何首乌炮制减毒作用机制可能与调节这些代谢途径相关,以期为何首乌炮制减毒提供科学依据。
Based on the model of immune stress, the mechanism of Metabolic mechanism of HeLuo Wu Jing (JiuShuiJiuYanJiu) after its preparation was studied. Polygonum multiflorum and 50% ethanol extract of Radix Polygonum multiflorum were administered alone or in combination with non-toxic dose of lipopolysaccharide (LPS) to SD rats. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase ), The pathological changes of liver were observed by HE staining, and the toxicity differences of Polygonum multiflorum Thunb. The metabolic profiles of plasma samples of different groups were analyzed by UPLC-QTOF / MS. Combined with partial least-squares discriminant analysis (OPLS-DA) To investigate the effect of endogenous metabolites in the plasma of rat with heterogeneous liver injury and to find out the potential biomarkers of Polygonum multiflorum Thunb., Which are attenuated, and to import the identified biomarkers into the KEGG database to construct metabolic pathways. The results showed that there was no significant change of ALT and AST in rats before and after the single gavage of Radix polygoni multiflori (5.4 g · kg -1), no obvious pathological changes were found in the liver slices, while the same dose of Polygonum multiflorum After administration of LPS, the ALT and AST of Polygonum multiflorum Thunb. Group were significantly increased (P <0.01), and obvious liver pathological changes were observed in the liver slices. Principal component analysis (PCA) results showed that the serum metabolites profiles of normal control group, LPS group, LPS / Shouwu group and LPS / Polygonum multiflorum were significantly separated, and 10 potential biomarkers associated with liver injury were found and identified . It is speculated that these biomarkers may be related to sphingolipid metabolism, linoleic acid metabolism, steroid hormone biosynthesis, galactose metabolism, steroid biosynthesis, cytochrome P450 exogenous substance metabolism, pyrimidine metabolism, unsaturated fatty acid synthesis, primary bile acid Synthesis, and taurine and hypotaurine metabolism and other 10 metabolic pathways. So Polygonum multiflorum engineered attenuating mechanism may be related to regulating these metabolic pathways, in order to provide a scientific basis for why Radix Polygoni multiflorum processing attenuated.