ABSTRACT
Objective:To characterize the structure of polysaccharide isolated from Linggui Zhugan Tang（LGZGT）,including monosaccharide composition and functional group detection, investigate the difference of the antioxidant activities of crude polysaccharide（CP） and pure polysaccharide（PP）, and provide the basis for the quality evaluation of LGZGT by in vitro bioassay. Method:The average molecular weight of CP was analyzed by high performance gel chromatography（HPGPC）. Gas chromatography-mass spectrometry（GC-MS） and fourier transform infrared（FT-IR） were employed to determine the structure of the polysaccharide. The antioxidant activities of CP and PP samples were evaluated on the basis of 1-diphenyl-2-picrylhydrazyl（DPPH） radical scavenging activity and OH radical scavenging activity. Result:The total polysaccharide was composed of single peaks, with a molecular weight of 3 689 Da. It was mainly composed of arabinose, mannose, glucose, galactose and fructose with a molar ratio of 6.85∶1.00∶109.21∶1.04∶21.82. Among them，glucose and fructose were the predominant components. In addition， IR study indicated the presence of pyranose and anomeric configurations in glycan structure, with two stereoisomers of glycosidic bond (α-glycosidic bond and β-glycosidic bond). It was found that the total polysaccharide had the ability of scavenging DPPH and hydroxyl radicals, and the activity of crude polysaccharide was better than that of refined polysaccharide. It was found in antioxidant research that the total polysaccharide had the ability of scavenging DPPH and hydroxyl radicals, and the activity of CP was better than that of PP. Furthermore, LC-Q-TOF-MS was used to qualitatively analyze the other components in CP, which indicated that it was related to the adsorption of pentacyclic triterpenoids in Glycyrrhiza uralensis. Conclusion:The polysaccharides and pentacyclic triterpenoids in LGZGT are the material basis for the antioxidative effect of LGZGT. The antioxidative activity determined by in vitro bioassay can be used as an evaluation index for the overall quality control of LGZGT.
ABSTRACT
Linggui Zhugan Tang(LGZG), a representative of warming spleen to resolve fluid retention, is widely used in various diseases caused by spleen deficiency and fluid retention (SDFR). Ancient doctors mostly focused on the theme of SDFR to analysis the composition of LGZG. Modern research has now found that most of the components of LGZG contain anti-inflammatory, anti-tumor and immune-modulating effects. At present, the pharmacological research of LGZG mainly involves clearance of β-amyloid (Aβ) and anti-neuroinflammation, regulation of lipid metabolism and insulin resistance, protection of myocardial cells and regulation of liquid metabolism as follow. LGZG can promote the clearance of Aβ by regulating the expression of Aβ transporters, including low-density lipoprotein receptor-associated protein-1 and terminal glycosylation product receptors, as well as reduce the inflammation of nerve cells by inhibiting the secretion of pro-inflammatory factors such as interleukin(IL) -1β,IL-6 and tumor necrosis factor-α(TNF-α) by microglia. The prescription can also affect the lipid metabolism and insulin resistance by regulating the levels of adiponectin, insulin and leptin, thus producing therapeutic effects on metabolic syndrome and non-alcoholic fatty liver disease. The protective mechanism of LGZG on myocardial cells mainly contains inhibition of the over-activation of nuclear transcription factor-κB(NF-κB) signaling pathway,up-regulation of Smad7 expression and down-regulation of Smad3 expression, regulation of inhibitor of nuclear factor kappa-B kinase (IKK)/NF-κB inhibitor (IκB)/NF-κB signaling pathway, inhibition of inflammatory factor production, and down-regulation of Caspase-3 and Caspase-8 expression. In addition,LGZG can also regulate aquaporins to affect water metabolism. And it also has significant antioxidant effects. As is known to all,the functional mechanism of traditional Chinese medicine (TCM) compound is closely related to the network regulation of TCM disease syndromes. Considered from this angle, it is helpful to carry out the "disease-syndrome-formula" research by means of network pharmacology characterized by "disease-target-drug", thus organically exploring the relationship between LGZG and SDFR from the microscopic perspective,and helping to explain the scientific connotation of SDFR and the biological mechanism of warming spleen to resolve fluid retention.
ABSTRACT
Objective:To observe the effect of Linggui Zhugan Tang on gut microbiota in ob/ob mice, in order to explore whether its bone protective effect is related to its regulatory effect of gut microbiota. Method:The ob/ob mice were divided into control group, model group, Linggui Zhugan Tang group, and Atorvastatin group. After administration for 4 weeks, morphological changes in intestine and bone were observed by htoxylin eosin(HE) staining. The protein expression of alkaline phosphatase(ALP) was detected by Western blot in bone. The expression of farniol X receptor(FXR) was detected by in immunofluorescence staining. And the content of interleukin-6(IL-6) in serum was analyzed by the enzyme-linked immunosorbent assay(ELISA) method. Gut microbiota was analyzed by 16S rRNA. Result:Compared with control group, the trabecular bone of the femur became thinner, the bone marrow cavity was infiltrated with fat, and the epithelium of the small intestine was damaged in the model group. The protein expressions of ALP and FXR decreased, while the content of IL-6 in serum was increased in the model group. Compared with model group, the protein expressions of ALP and FXR increased, while the content of IL-6 in serum was decreased in the Linggui Zhugan Tang group. At the genus level, compared with control group, the abundance of Lachnospiraceae_UCG-001 increased (PBacteroidales_S24-7_group decreased in model group (PLachnospiraceae_UCG-001 was positively correlated with the weight of body (r=0.661), while norank_f__Bacteroidales_S24-7_group was negatively correlated with the weight of body (r=-0.622). Conclusion:Linggui Zhugan Tang has a protective effect on bone injury in ob/ob mice, and its mechanism may be related to its regulation of gut microbiota.