A feasible,economical,and accurate analytical method for simultaneous determination of six alkaloid markers in Aconiti Lateralis Radix Praeparata from different manufacturing sources and processing ways

Aconiti Lateralis Radix Praeparata(Fuzi) is a commonly used traditional Chinese medicine in clinic for its potency in restoring yang and rescuing from collapse. Aconiti alkaloids, mainly including monoester-diterpenoidaconitines(MDAs) and diester-diterpenoidaconitines(DDAs), are considered to act as both bioactive and toxic constituents. In the present study, a feasible, economical, and accurate HPLC method for simultaneous determination of six alkaloid markers using the Single Standard for Determination of Multi-Components(SSDMC) method was developed and fully validated. Benzoylmesaconine was used as the unique reference standard. This method was proven as accurate(recovery varying between 97.5%-101.8%, RSD < 3%), precise(RSD 0.63%-2.05%), and linear(R > 0.999 9) over the concentration ranges, and subsequently applied to quantitative evaluation of 62 batches of samples, among which 45 batches were from good manufacturing practice(GMP) facilities and 17 batches from the drug market. The contents were then analyzed by principal component analysis(PCA) and homogeneity test. The present study provided valuable information for improving the quality standard of Aconiti Lateralis Radix Praeparata. The developed method also has the potential in analysis of other Aconitum species, such as Aconitum carmichaelii(prepared parent root) and Aconitum kusnezoffii(prepared root).

Study on Hot Property of Aconiti Lateralis Radix Praeparata and Its Compatibility with Zingiberis Rhizoma Based on Animal Temperature Tropism

Objective To establish an objective method for evaluating the intrinsic characteristics between hot and cold properties of herbal drugs and to study the hot property of Aconiti Lateralis Radix Praeparata(ALRP) and its compatibility with Zingiberis Rhizoma(ZR) on animal temperature tropism.Methods The equipment with cold/hot pads was used to investigate the variety of temperature tropism among mice treated by ALRP and its compatibility with ZR.Meanwhile,the activities of adenosine triphosphatase(ATPase),total anti-oxidant capability(T-AOC),and total superoxide dismutase(T-SOD) were measured.Results Compared with the homologous ALRP group,the compatibility between ALRP and ZR(AZ) had a stronger action against the decreasing of remaining rate on hot pad(P < 0.05),suggesting an enhancement of cold tropism.Meanwhile,the internal indexes of Na+-K+-ATPase activity,Ca2+-Mg2+-ATPase activity,and oxygen consuming volume increased significantly(P < 0.05),suggesting an enhancement of energy metabolism.The changes of T-AOC and T-SOD activity suggested that AZ reinforced the anti-oxidative capability of mice.In addition,it could be inferred from cluster analysis that the activity of AZ was concretely stronger than that of homologous ALRP.Conclusion ALRP and ZR are both herbs with hot property,but AZ has an enhanced trend of hot property.This external behavior of cold tropism and the internal activity of energy metabolism and anti-oxidant might reflect the internal hot property in an intuitive and objective way.The changes of ATPase activity of liver tissue might be the mechanism of drug action.

Research Progress in Toxicity and Toxicity-reducing and Efficacy-enhancing Compatibility of Radix Aconiti Lateralis Praeparata

Radix Aconiti Lateralis Praeparata contains alkaloids,flavonoids,polysaccharides,organic acids and organic alkalis,among which diester alkaloids are important effective ingredients,which can be used for treating heart failure,rheumatic heart disease,coronary heart disease,hypotension,shock and other diseases.However,it also contains the main ingredients that produce toxic effects and are highly toxic to the cardiovascular system,nervous system,digestive system,reproductive system,embryos,and livers and kidneys.In this study,we analyzed the toxicity-reducing and efficacy-enhancing effects of the compatibility of Radix Aconiti Lateralis Praeparata with with single herbs Glycyrrhiza uralensis Fisch.,Panax ginseng C.A.Meyer,Zingiber ojjicinale Rosc.and Rheum palmatum L.,aiming to provide references for further exploring the appropriate compatibility conditions of traditional Chinese medicine in the future and improving the safety of the clinical application of Radix Aconiti Lateralis Praeparata.

Synergetic effects of aqueous extracts of Fuzi(Radix Aconiti Lateralis Preparata) and Tubeimu(Rhizoma Bolbostemmatis) on MDA-MB-231 and SKBR3 cells

OBJECTIVE： To test the synergistic effects of the aqueous extract of Tubeimu （Rhizoma Bolbosternmatis） and Fuzi （Radix Aconiti Lateralis Preparata） on MDA-MB-231 and SKBR3 breast cancer cells.METHODS： A combined index was created for the effects of Tubeimu （Rhizoma Bolbostemmatis） and Fuzi （Radix Aconiti Lateralis Preparata） extracts. Cell proliferation was performed by trypan blue exclusion and 3-（4,5-dimethylthiazol-2-yl）-5-（3- carboxy- methoxyphenyl）-2-（4-sulfophenyl）-2H-tetrazolium （MTS） assays. Flow cytometry was used to assess cell cycle distribution and apoptosis. Cell migration was determined by wound-healing and transwell assays. Confocal microscopy was used to detect E-cadherin and actin filaments. RESULTS： The aqueous extract from Tubeimu （Rhizoma Bolbostemmatis）and Fuzi （Radix Aconiti lateralis Preparata） exerted synergetic effects on the growth of MDA-MB-231 cells and G1 phase arrest. When exposed to extracts at concentrations of 62.5 ：62.5 and 62.5：31.3 μg/mL, the combination index was 0.83 and 0.74, respectively. Interestingly, 62.5： 31.3 μg/mL of combined drugs enhanced the inhibitory effect of Tubeimu （Rhizoma Bolbostemmatis） on the migration of SKBR3 cells and reduced the stimulative effect of Fuzi （Radix Aconiti Lateralis Preparata） （P 〈 0.01）, in which cells showed an increased expression of E-cadherin and reorganization of actin filaments （P 〈 0.001）. 62.5：62.5 μg/mL extract also synergistically induced apoptosis of MDA-MB-231 cells （P 〈 0.05 and P 〈 0.001）. Acting as the main active ingredients in the extract, tubeimoside I and acetylbenzoylaconine at 10：10μg/ mL and 5：2.5 μg/mL also produced inhibitory effects on the proliferation and migration of cells （P 〈 0.01）. CONCLUSION： Tubeimu （Rhizoma Bolbostemmatis） and Fuzi （Radix Aconiti Lateralis Preparata） extracts had synergic effects on MDA-MB-231 and SKBR3 cells.

Effects of Radix Aconiti Lateralis Preparata and Rhizoma Zingiberis on Energy Metabolism and Expression of the Genes Related to Metabolism in Rats

Objective： To study the influence of Radix aconiti lateralis preparata and Rhizoma zingiberis, two species of Chinese medicinal herbs with hot property, on energy metabolism and gene expression spectrum, and to analyze the possible mechanism of their effects. Methods： Forty-eight specific pathogen free Wistar rats were randomly divided into a Radix aconiti lateralis preparata group, a Rhizoma zingiberis group, and a control group. They were intragastrically treated with concentrated decoction of Radix aconiti lateralis preparata, Rhizoma zingiberis and normal saline respectively for 20 days. Toe temperature （TT）, energy intake （El）, digestible energy （DE）, and metabolizable energy （ME） were measured. The content of adenosine triphosphate （ATP） and energy charge （EC） in hepatic tissue were measured with high performance liquid chromatography （HPLC）. The activity of ATPase and succinate dehydrogenase （SDH） in the liver were detected with chemical colorimetry. The gene expression in the liver was detected with Illumina＇s rat Ref-12 gene array. The differential expression genes were selected, annotated and classified based on Gene Ontology （GO）. Real-time quantitative reverse-transcriptase PCR （Q-RT-PCR） was used to test the accuracy of results. Results： Compared with the control group, the -I-r on the 10^th day after the beginning of administration and ATP in the Radix aconiti lateralis preparata and Rhizoma zingiberis groups increased significantly （P〈0.05）. El/body mass （BM）, DE/BM, ME/BM, the hepatic EC and the activity of Na^＋-K^＋-ATPase, Ca^2＋-Mg^2＋-ATPase and SDH of liver increased significantly only in the Radix aconiti lateralis preparata group （P〈0.05）. There were 592 differential expression genes in the Radix aconiti/ateralis preparata group and 1 159 in the Rhizoma zingiberis group compared with the control group. Among the differential expression genes, genes related to metabolic processes were the most significant based on GO analysis. There were 337 strips of gene differential expression in common in both Radix aconiti lateralis preparata and Rhizoma zingiberis groups compared with the control group. Conclusions： Herbs with hot property such as Radix aconiti lateralis preparata and Rhizoma zingiberis could improve the energy metabolism in rats, through influencing the metabolic process of sugar, lipid, and amino acid. It could also promote the production, storage, and utilization of energy by regulating the gene expression related to metabolism, which may be the main molecular mechanism of warming yang and dispelling cold for the treatment of the cold syndrome according to Chinese medicine theory.

Efficacy of Renshen(Radix Ginseng) plus Fuzi(Radix Aconiti Lateralis Preparata) on myocardial infarction by enhancing autophagy in rats

OBJECTIVE:To elucidate the protective effects of Renshen(Radix Ginseng)and Fuzi(Radix Aconiti Lateralis Preparata)on myocardial infarction(MI)through regulating myocardial autophagy.METHODS:Thirty-one male Sprague-Dawley rats were randomized into five groups(n=6 or 7 for each).After treatment for 3 weeks,electrocardiogram(ECG)and cardiac function were recorded.Hematoxylin and eosin(HE)staining was used to detect pathological changes in the heart.Enzyme-linked immunosorbent assay(ELISA)was used to detect serum B-type brain natriuretic peptide(BNP),cardiac troponin T(c Tn T),tumor necrosis factor-α(TNF-α),and serum inflammatory cytokines.Metabolomic analysis was used to identify differential biomarkers of MI in rats.Immunohistochemistry and western blotting were used to detect BNP,cTnT,TNF-α,LC3B,Beclin-1,p62,and adenosine monophosphate activated protein kinase(AMPK)expression in cardiac tissue.RESULTS:Fuzi(Radix Aconiti Lateralis Preparata)alone or Renshen(Radix Ginseng)plus Fuzi(Radix Aconiti Lateralis Preparata)markedly ameliorated cardiac dysfunction and abnormal ECGs,demonstrated by decreases in the heart weight/body weight ratio,BNP,and c Tn T.Pro-inflammation cytokine interleukin(IL)-1αsignificantly decreased and anti-inflammatory cytokine IL-10 significantly increased in Renshen(Radix Ginseng)single or Renshen(Radix Ginseng)plus Fuzi(Radix Aconiti Lateralis Preparata)groups compared with the control group.HE results suggested that co-treatment produced a greater reduction in cardiomyocyte cross-sectional area than Renshen(Radix Ginseng)or Fuzi(Radix Aconiti Lateralis Preparata)alone.Renshen(Radix Ginseng)plus Fuzi(Radix Aconiti Lateralis Preparata)reversed these changes to different degrees in MI rats.Furthermore,Renshen(Radix Ginseng)plus Fuzi(Radix Aconiti Lateralis Preparata)down-regulated LC3 B,Beclin-1,and AMPK expression in cardiac tissue and upregulated p62 expression.CONCLUSIONS:Renshen(Radix Ginseng)plus Fuzi(Radix Aconiti Lateralis Preparata)may have a greater effect on heart injury induced by MI in rats than Fuzi(Radix Aconiti Lateralis Preparata)treatment alone,and the underlying mechanism may be associated with the regulation of myocardial autophagy and anti-inflammation effects.These results provide fresh insight into the mechanism of co-treatment with Renshen(Radix Ginseng)and Fuzi(Radix Aconiti Lateralis Preparata)for MI.

Inhibitory effect of Sumu(Lignum Sappan) plus Fuzi(Radix Aconiti Lateralis Praeparata) on a lung carcinoma model

OBJECTIVE: To investigate the inhibitory effect of Sumu(Lignum Sappan) plus Fuzi(Radix Aconiti Lateralis Praeparata)(SF) on the growth and metastasis of Lewis lung carcinoma.METHODS: A lung carcinoma model was established by subcutaneously inoculating Lewis lung carcinoma cells into C57 BL/6 mice. The mice were randomly divided into four groups of 13 mice(control, low-dose, moderate-dose, and high-dose), and gavaged once-daily for 21 consecutive days. The rates of tumor inhibition, metastasis, and metastasis inhibition were observed. The differential expressions of s P-selectin and vascular endothelial growth factor C(VEGFC) were compared between the treatment groups and the control group.RESULTS: The tumor weights differed significantly between the treatment groups and the control group(P < 0.05). Administration of SF in the moderate-dosage and low-dose groups significantly inhibited the expression of s P-selectin and VEGFC(both P < 0.05), suggesting anti-tumor activity.CONCLUSION: SF can inhibit the growth and metastasis of Lewis lung carcinoma.

Comparative study of anti-inflammatory effects of different processed products through the COX-2/PGE2 signaling pathway: based on network pharmacology and molecular docking

Background:Radix Aconiti Lateralis Preparata(Fu-zi)is a traditional Chinese medicinal herb,which has been widely used in the clinic and has potent anti-inflammatory activities.we aimed to explore the mechanisms of extract containing alkaloids from different Fu-zi Processed Products(FPP)in treating inflammation,especially rheumatoid arthritis(RA).Methods:Firstly,using network pharmacology technology,the ingredients,and targets of Fu-zi were obtained by searching and screening,the targets involving RA were acquired,the intersection targets were constructed a"component-target-pathway"network.A comprehensive investigation was conducted on the anti-rheumatoid arthritis mechanisms of 5 FPPs in lipopolysaccharide(LPS)induced RAW264.7 cells,which serve as a model for RA.The production of NO and inflammatory cytokines were measured by ELISA kit.Quantitative Real-time PCR(qRT-PCR)was utilized to measure the mRNA levels.COX-2/PGE2 signaling pathway-associated proteins were determined by western blot.Results:According to a network pharmacological study,16 chemical components and 43 common targets were found in Fu-zi and 6 key targets including PTGS2 were closely related to the mechanism of Fu-zi in treating RA.The in vitro study revealed that the levels of NO,TNF-α,and IL-1βwere substantially decreased by the 5 FPPs.The 5 FPPs significantly suppressed the expression of proteins COX-2,iNOS,and NF-κB,with particularly notable effects observed for PFZ and XFZ.Conclusion:Altogether,these results demonstrated that the 5 PPS containing alkaloids have a good anti-RA-related inflammatory effect,and the mechanism may be related to COX-2/PGE2 signaling pathway,particularly,Fu-zi prepared utilizing a traditional Chinese technique.

Relationship between cardiotonic activity of Fuzi(Radix Aconiti Lateralis Preparata)and its fingerprint determined by liquid chromatography-mass spectrometry

OBJECTIVE:To investigate the relationship between the cardiotonic activity of Fuzi(Radix Aconiti Lateralis Preparata,RALP)and its fingerprint determined by liquid chromatography-mass spectrometry(LC-MS).METHODS:First,the fingerprints of six processed products of RALP were established by high performance liquid chromatography quadrupole time-of-flight mass spectrometry(HPLC-Q-TOF-MS)followed by analysis of the principal component of the relative peak area of its common peaks.Next,the scores of the first five principal components were used as input for an artificial neural network(ANN).Additionally,the therapeutic effect of RALP was assessed by measuring the hemodynamic indexes of heart failure model rats.Subsequently,fluorescence semi-quantitative polymerase chain reaction and an enzyme-linked immunosorbent assay kit were used to determine the effects of RALP-processed products on the serum levels of noradrenaline(NA),angiotensin-Ⅰ(Ang-Ⅰ),and the expression ofβ-norepinephrine receptor m RNA(β-NRm)in the rat cardiac tissues.P<0.05 was used as the output of the ANN.Finally,a network was constructed to display the relationship between the LC-MS fingerprints and the cardiotonic activity of the RALP-processed products.RESULTS:Several types of RALPs can improve diastolic function,systolic function and heart rate.On the basis of the findings from the principal component analysis(PCA)of 16 common peaks of fingerprints of six RALP-processed products,it was revealed that the first five principal components may include 100%of the information of the original data.As observed from the multilayer perceptron neural network analysis,principal component 4 presented with the strongest effects on serum levels of NA and Ang-Ⅰin rats,while principal component1 exerted the greatest effect onβ-NRm expression in cardiac tissue.CONCLUSION:The key findings obtained from this study indicated that the network constructed by the PCA-ANN may predict pharmacodynamic effects of the main ingredients of Traditional Chinese Medicine(TCM).This method may serve as a new approach to identify the relationship between LC-MS fingerprints and the pharmacodynamic effects of TCM ingredients.

Metabolomics of Fuzi-Gancao in CCl4 induced acute liver injury and its regulatory effect on bile acid profile in rats

BACKGROUND Fuzi(Radix aconiti lateralis)-Gancao(Radix glycyrrhizae)is one of the most classical drug pairs of traditional Chinese medicine.In clinical practice,decoctions containing Fuzi-Gancao(F-G)are often used in the treatment of liver diseases such as hepatitis and liver failure.AIM To investigate the metabolomics of F-G in CCl4 induced acute liver injury in rats and its regulatory effect on the bile acid profile.METHODS The pharmacodynamic effect of F-G on CCl4 induced acute liver injury in rats was evaluated,and an ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method for the simultaneous determination of 92 metabolites from multiple pathways was established to explore the protective metabolic mechanism of F-G in serum on the liver.RESULTS Twenty-four differential metabolites were identified in serum samples.The primary bile acid biosynthetic metabolic pathway was the major common pathway in the model group and F-G group.Subsequently,a UPLC-MS/MS method for simultaneous determination of 11 bile acids,including cholic acid,ursodeoxycholic acid,glycochenodeoxycholic acid,glycochenodeoxycholic acid,taurocholic acid,glycocholic acid,chenodeoxycholic acid,deoxycholic acid,taurochenodeoxycholic acid,taurocholic acid,and glycinic acid,was established to analyze the regulatory mechanism of F-G in serum.F-G decreased the contents of these 11 bile acids in serum in a dose-dependent manner compared with those in the model control group.CONCLUSION F-G could protect hepatocytes by promoting the binding of free bile acids to glycine and taurine,and reducing the accumulation of free bile acids in the liver.F-G could also regulate the compensatory degree of taurine,decreasing the content of taurine-conjugated bile acids to protect hepatocytes.

Pharmacological effects of Chinese herb aconite(Fuzi) on cardiovascular system

Fuzi(aconite,Radix Aconiti praeparata),a widely used Chinese herb,plays a significant role in the cardiovascular system.This is mainly reflected by Fuzi's cardiotonic effect,its protective effect on myocardial cells,and its effect on heart rate and rhythm,blood pressure,and hemodynamics.In this article,the pharmacological effects and the corresponding mechanisms of Fuzi(aconite) and its active components on cardiovascular system are reviewed.