苍术多糖与正丁醇部位对脾虚大鼠线粒体自噬的影响  

Effects of polysaccharides and n-butanol fractions of Atractylodis Rhizoma on mitophagy in rats with spleen deficiency

作　　者：徐慧颖 丁奕辉 董昊 赵雪岑 柏阳 才谦[1] XU Hui-ying;DING Yi-hui;DONG Hao;ZHAO Xue-cen;BAI Yang;CAI Qian(College of Pharmacy,Liaoning University of Traditional Chinese Medicine,Dalian 116600,China)

机构地区：[1]辽宁中医药大学药学院,大连116600

出　　处：《天然产物研究与开发》2023年第8期1372-1379,共8页Natural Product Research and Development

基　　金：国家自然科学基金(81973478);兴辽英才计划(XLYC2002004)。

摘　　要：多糖与正丁醇部位是苍术的健脾活性部位,本文从线粒体自噬的角度探讨苍术活性部位对脾虚大鼠的调节作用,进而阐释其健脾机制。采用透射电镜观察大鼠胃窦组织线粒体超微结构;采用qRT-PCR和Western blot法检测大鼠胃窦组织线粒体PINK1、Parkin、LC3和P62 mRNA及蛋白表达水平;采用比色法检测大鼠胃窦组织三磷酸腺苷(adenosine-triphosphate,ATP)、H_(2)O_(2)水平,JC-1荧光探针法检测线粒体膜电位(MMP)水平。结果表明,与模型组相比,麸炒苍术多糖(FD)、麸炒苍术正丁醇部位(FZ)、生苍术多糖(SD)与生苍术正丁醇部位(SZ)均不同程度改善了脾虚大鼠胃窦组织线粒体结构损伤,且自噬溶酶体数量增多;FD、FZ、SD与SZ均能显著下调PINK1、Parkin、p62 mRNA与蛋白表达,上调LC3II蛋白表达、LC3II/I值;FD与FZ组ATP、MMP水平显著上升,H_(2)O_(2)水平显著下降,SZ组ATP、MMP水平显著上升,SD组H_(2)O_(2)水平显著下降。综上所述,苍术多糖与正丁醇部位可能是通过改善自噬障碍,恢复线粒体功能,从而调节脾虚,且麸炒后多糖与正丁醇部位效果明显优于生品。Polysaccharides and n-butanol fractions are effective fractions in Atractylodis Rhizoma(AR)for treating spleen deficiency.The study is to explore the mechanism of the effective fractions in AR on treating spleen deficiency from the perspective of mitophagy.The ultrastructure of mitochondria in gastric antrum of rats was observed by transmission electron microscope.The mRNA and protein expression levels of autophagy-related proteins PINK1,Parkin,LC3 and p62 were detected by qRT-PCR and Western blot.The adenosine triphosphate(ATP)and H_(2)O_(2) levels in gastric antrum of rats were detected by biochemical analysis,the level of mitochondrial membrane potential(MMP)were detected by JC-1.The results showed that compared with the model group,the polysaccharides of bran-processed AR(FD),n-butanol fractions of bran-processed AR(FZ),polysaccharides of crude AR(SD)and n-butanol fractions of crude AR(SZ)improved the mitochondrial structure injury and increased the number of autophagic lysosomes in different degrees.FD,FZ,SD and SZ significantly down-regulated the expression of PINK1,Parkin and p62 mRNA and protein,and up-regulated the expression of LC3II protein and LC3II/I value;the levels of ATP and MMP in FD and FZ groups increased significantly,while the levels of H_(2)O_(2) decreased significantly,the levels of ATP and MMP in SZ group increased significantly,while the levels of H_(2)O_(2) in SD group decreased significantly.In summary,the polysaccharides and n-butanol fractions of AR may treat spleen deficiency by promoting mitophagy and improving mitochondrial function of gastric antrum,and the effect of polysaccharides and n-butanol fractions of bran-processed AR were obviously superior to that of crude AR.

关 键 词：苍术 麸炒 脾虚 多糖 正丁醇部位 

分 类 号：R285[医药卫生—中药学] R932[医药卫生—中医学]