ABSTRACT
Objective:To detect the toxicity of water-eluted fraction from Siegesbeckiae Herba （SWEF） at different concentrations against MRC-5 human embryonic lung fibroblasts and its impacts on the expression of <italic>α</italic>7 nicotinic acetylcholine receptor （<italic>α</italic>7nAChR） and inflammatory factors， so as to figure out the active components responsible for toxicity and efficacy. Method:The toxicities of SWEF at 1， 6， 10， 20， and 50 g·L<sup>-1</sup> against MRC-5 cells were determined by cell counting kit-8 （CCK-8） assay combined with flow cytometry and Trypan blue staining. The changes in <italic>α</italic>7nAChR expression and inflammatory factor levels before and after <italic>α</italic>7nAChR gene silencing were detected to reveal the pharmacodynamic effect of SWEF on MRC-5 cells. Result:SWEF （≥6 g·L<sup>-1</sup>） obviously inhibited the viability of MRC-5 cells （<italic>P</italic><0.01） and promoted their apoptosis and necrosis （<italic>P</italic><0.01）， with the half-maximal inhibitory concentration （IC<sub>50</sub>） being 6.03 g·L<sup>-1</sup>. The determination of <italic>α</italic>7nAChR expression and inflammatory factor levels in MRC-5 cells showed that SWEF contained <italic>α</italic>7nAChR agonist-like substance， which enhanced <italic>α</italic>7nAChR mRNA and protein expression （<italic>P</italic><0.05， <italic>P</italic><0.01） and decreased the inflammatory factor levels （<italic>P</italic><0.05， <italic>P</italic><0.01）. SWEF down-regulated the inflammatory factors possibly by re-regulating <italic>α</italic>7nAChR mRNA expression， exhibiting a negative correlation between them （<italic>P</italic><0.01）. Conclusion:SWEF （≥6 g·L<sup>-1</sup>） is highly toxic to MRC-5 cells. Pharmacodynamic studies have confirmed that <italic>α</italic>7nAChR agonist-like substance contained in SWEF was responsible for the elevated <italic>α</italic>7nAChR expression and reduced inflammatory cytokines. It is inferred that excessive <italic>α</italic>7nAChR agonist-like substance may trigger the toxicity of<italic> </italic>Siegesbeckiae Herba.
ABSTRACT
Objective:Metabolomics was used to analyze the dynamic changes of endogenous metabolites in serum and lung tissue of rats treated with water elution section of Siegesbeckiae Herba (SWES), and to explore the possible mechanism of lung injury and to search for the sensitive markers in serum and lung tissue. Method:SD rats were randomly divided into three groups, namely the normal group (normal saline), SWES high-dose group and SWES low-dose group (0.500, 0.125 g·kg-1·d-1). SWES was given for 4 weeks and stopped for 2 weeks. The weight and pathological examination were regarded as observation parameters. Serum and lung tissue samples were detected by nuclear magnetic resonance hydrogen spectrum (1H-NMR). The metabolites in rats were analyzed by partial least squares-discriminant analysis (PLS-DA) and orthogonal partial least squares-discriminant analysis (OPLS-DA). Result:Lung inflammation was shown in SWES high-dose group and returned to normal after withdrawal for 2 weeks. The metabolic spectrum of SWES high-dose group was significantly different from the normal group. There were 11 metabolites were identified by 1H-NMR metabolomics, four differential metabolites were identified in serum [acetate, trimethylamine oxide (TMAO), glycine, myo-inositol] and nine differential metabolites were identified in lung tissue [lactate, acetate, phosphatidylcholine (PC), pyruvate, dimethylamine, glutamate, glycerophosphocholine (GPC), glycine, xanthine]. Lung injury was related to the disorder of pyruvate metabolism, glutamate metabolism and other pathways. Most of the metabolites in lung tissue had obviously levels of callback after drug withdrawal, which coincided with the pathological examination. Conclusion:The lung is one of the damaged organs caused by SWES, and the lung injury is reversible and may be related to energy metabolism and oxidative stress.
ABSTRACT
Xixiancao( Siegesbeckiae Herba) has the effect of treating ischemic stroke( IS),however,the mechanism has not been fully elucidated. In this study,combined with Lipinski's five principles and Veber oral bioavailability rules,68 chemical components of Xixiancao were obtained by database and literature search. Based on the reverse targeting,248 potential targets were obtained and mapped it to the ischemic stroke target set,47 potential targets for the treatment of ischemic stroke were obtained. Molecular docking technique was used to verify that the Xixiancao component has good binding activity to potential targets. GO enrichment analysis and pathway analysis were performed on potential targets using Clue GO. GO enrichment analysis showed that Xixiancao was mainly involved in life processes such as neuronal apoptosis,cholesterol storage and blood pressure regulation. Pathway analysis showed that Xixiancao may promote vascular repairing and regeneration by regulating the expression of ADAMTS1,FLT1 and KDR in VEGFA-VEGFR2 signaling pathway,activate cell survival signals and inhibit neuronal apoptosis by regulating the expression of CAMK2 AA,MDM2,MAPK1,MAPK3,CDK5 and MAPK10 in brain-derived neurotrophic factor signaling pathway and PI3 K-Akt signaling pathway. Lipid homeostasis and inflammation may also be regulated by Xixiancao through regulating the expression of ESR1,NR1 H3,PPARA,PPARG in the nuclear receptor signaling pathway. In addition,Xixiancao could also prevent platelet aggregation by regulating the expression of ITGA2 B,F2,F10,and ALB,and play an antithrombotic role. The results of this study indicate that Xixiancao plays an important role in the treatment of ischemic stroke mainly through anti-thrombosis,promoting angiogenesis,protecting neurons,anti-inflammatory and regulating blood pressure and lipids.
Subject(s)
Humans , Asteraceae , Chemistry , Brain Ischemia , Drug Therapy , Drugs, Chinese Herbal , Therapeutic Uses , Molecular Docking Simulation , Signal Transduction , Stroke , Drug TherapyABSTRACT
OBJECTIVE:To explore the effects of extracts from Siegesbeckiae herba (HS) on doxorubicin (DOX)-induced chronic myocardial injury in mice. METHODS:48 mice were randomly divided into 3 groups. Mice in blank control(Con)group received distilled water once every day,ig,and normal saline(2 mL/100 g)once every day,ip,for 8 weeks;mice in DOX mice received distilled water(2 mL/100 g)once every day,ig,and DOX(3 mg/kg)once every week,ip,for 8 weeks;mice in DOX+HS group received HS(340 mg/kg)once every day,ig,and DOX(3 mg/kg)once every week,ip,for 8 weeks. After administra-tion,body mass,heart coefficient,cardiac function changes,serum biochemical index levels [alanine aminotransferase(ALT),as-partate aminotransferase (AST), creatine phosphokinase (CK), creatine kinase isoenzyme (CK-MB), lactate dehydrogenase (LDH)and total cholesterol(TC)] of mice were determined. RESULTS:Compared with Con group,body mass of mice in DOX group was decreased(P<0.01);heart coefficient was increased(P<0.05);heart rate slowed down,R-wave was increased(P<0.05 or P<0.01);serum biochemical indexed were increased,there was significant difference in AST(P<0.01). Compared with DOX group,heart coefficient of mice in DOX+HS group was decreased (P<0.01);heart rate was increased (P<0.01);serum biochemical indexes were decreased,there was significant differences in CK,LDH,TC(P<0.01). CONCLUSIONS:HS has cer-tain protective effects on DOX-induced chronic myocardial injury in mice.