Yanling Zhao

Comparative pharmacokinetic study of paeoniflorin and albiflorin after oral administration of Radix Paeoniae Rubra in normal rats and the acute cholestasis hepatitis rats.

A comparative study was designed and conducted to compare the pharmacokinetic difference of paeoniflorin and albiflorin after oral administration of Radix Paeoniae Rubra to normal rats and the acute cholestasis hepatitis rats induced by alpha-naphthylisothiocyanate (ANIT). UPLC-ESI-MS/MS method was employed to determine the level of paeoniflorin and albiflorin in rat plasma using geniposide as the internal standard (IS). Unpaired Students t-test was used for the statistical comparison. The investigation showed that there were significant differences between the normal rats and the acute cholestasis hepatitis rat groups in calculated parameters, such as AUC(0-t), AUC(0-∞), T(max) and CLz/F. The results indicated that acute liver injury in rats could alter the pharmacokinetics of drug. Since patients are the final users of the drug, it is essential to investigate the pharmacokinetics of the drug in disease status. Therefore, we used normal rats and the acute cholestasis hepatitis rats to study pharmacokinetics of Radix Paeoniae Rubra with the purpose of examining the influence of disease on the metabolic course.

Paeoniflorin alleviates liver fibrosis by inhibiting HIF-1α through mTOR-dependent pathway.

HIF-1α/mTOR signaling pathway is considered to play a crucial role in genesis and progress of tissue fibrosis. The elevation of HIF-1α and mTOR is relevant to CCl4 induced liver fibrotic rats. Paeoniflorin has been consistently shown to exhibit multiple pharmacological effects in liver disease. However, so far, no research demonstrates the relationship between paeoniflorin and HIF-1α/mTOR fibrogenesis pathway in liver fibrosis. In this study, the liver fibrosis was performed by CCl4 rats and HSC-T6 cell line. The data demonstrated that paeoniflorin treatment could attenuate liver fibrosis and inhibit the activation of HSC. Moreover, paeoniflorin significantly enhanced hepatic function by decreasing serum level of ALT, AST and ALP, and increasing level of ALB, TP. Meanwhile, ECM degradation was modulated by paeoniflorin treated rats with a remarkable reduce of α-SMA and collagen III mRNA expression. Moreover, the alleviation effect of liver fibrosis was relevant to inhibiting HIF-1α and phosphor-mTOR. Our data indicate that paeoniflorin alleviates liver fibrosis by inhibiting HIF-1α expression partly through mTOR pathway and paeoniflorin may be a potential therapeutic agent for liver fibrosis.

Paeonia lactiflora Pall. protects against ANIT-induced cholestasis by activating Nrf2 via PI3K/Akt signaling pathway

Background Paeonia lactiflora Pall. (PLP), a traditional Chinese herbal medicine, has been used for hepatic disease treatment over thousands of years. In our previous study, PLP was shown to demonstrate therapeutic effect on hepatitis with severe cholestasis. The aim of this study was to evaluate the antioxidative effect of PLP on alpha-naphthylisothiocyanate (ANIT)-induced cholestasis by activating NF-E2-related factor 2 (Nrf2) via phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway. Materials and methods Liquid chromatography-mass spectrometry (LC-MS) was performed to identify the main compounds present in PLP. The mechanism of action of PLP and its therapeutic effect on cholestasis, induced by ANIT, were further investigated. Serum indices such as total bilirubin (TBIL), direct bilirubin (DBIL), aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), γ-glutamyl transpeptidase (γ-GT), and total bile acid (TBA) were measured, and histopathology of liver was also performed to determine the efficacy of treatment with PLP. Moreover, in order to illustrate the underlying signaling pathway, liver glutathione (GSH) content and mRNA or protein levels of glutamate-cysteine ligase catalytic subunit (GCLc), glutamate-cysteine ligase modulatory subunit (GCLm), Akt, heme oxygenase-1 (HO-1), NAD(P)H/quinone oxidoreductase 1 (Nqo1), and Nrf2 were further analyzed. In addition, validation of PLP putative target network was also performed in silico. Results Four major compounds including paeoniflorin, albiflorin, oxypaeoniflorin, and benzoylpaeoniflorin were identified by LC-MS analysis in water extract of PLP. Moreover, PLP could remarkably downregulate serum levels of TBIL, DBIL, AST, ALT, ALP, γ-GT, and TBA, and alleviate the histological damage of liver tissue caused by ANIT. It enhanced antioxidative system by activating PI3K/Akt/Nrf2 pathway through increasing Akt, Nrf2, HO-1, Nqo1, GCLc, and GCLm expression. The putative targets network validation also confirmed the important role of PLP in activating Akt expression. Conclusion The potential mechanism of PLP in alleviating ANIT-induced cholestasis could to be related to the induction of GSH synthesis by activating Nrf2 through PI3K/Akt-dependent pathway. This indicates that PLP might be a potential therapeutic agent for cholestasis.

Serum Metabolomic Profiling in a Rat Model Reveals Protective Function of Paeoniflorin Against ANIT Induced Cholestasis.

Cholestasis is a leading cause of hepatic accumulation of bile acids resulting in liver injury, fibrosis, and liver failure. Paeoniflorin displays bright prospects in liver protective effect. However, its molecular mechanism has not been well‐explored. This study was designed to assess the effects and possible mechanisms of paeoniflorin against alpha‐naphthylisothiocyanate‐induced liver injury. Ultraperformance liquid chromatography coupled with quadrupole time‐of‐flight combined with principle component analysis and partial least squares discriminant analysis were integrated to obtain differentiating metabolites for the pathways and clarify mechanisms of disease. The results indicated that paeoniflorin could remarkably downregulate serum biochemical indexes and alleviate the histological damage of liver tissue. Different expression of 14 metabolites demonstrated that paeoniflorin mainly regulated the dysfunctions of glycerophospholipid metabolism and primary bile acid biosynthesis. Moreover, several pathways such as arginine and proline metabolism, ether lipid metabolism, and arachidonic acid metabolism were also related to the efficacy. In conclusion, paeoniflorin has indicated favorable pharmacological effect on serum biochemical indexes and pathological observation on cholestatic model. And metabolomics is a promising approach to unraveling hepatoprotective effects by partially regulating the perturbed pathways, which provide insights into mechanisms of cholestasis. Copyright

Paeoniflorin ameliorates ANIT-induced cholestasis by activating Nrf2 through an PI3K/Akt-dependent pathway in rats.

Cholestasis causes hepatic accumulation of bile acids leading to liver injury, fibrosis and liver failure. Paeoniflorin, the major active compound isolated from the roots of Paeonia lactiflora pall and Paeonia veitchii Lynch, is extensively used for liver diseases treatment in China. However, the mechanism of paeoniflorins hepatoprotective effect on cholestasis has not been investigated yet. In this study, we administered paeoniflorin to rats for 3u2009days prior to alpha‐naphthylisothiocyanate (ANIT) administration for once, then went on administering paeoniflorin to rats for 3u2009days. The data demonstrated that paeoniflorin significantly prevented ANIT‐induced change in serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphates (ALP), serum total bilirubin (TBIL), direct bilirubin (DBIL), total bile acid (TBA) and gamma‐glutamyl transpeptidase (γ‐GT). Histology examination revealed that paeoniflorin treatment rats relieved more liver injury and bile duct proliferation than ANIT‐administered rats. Moreover, our data indicated that paeoniflorin could restore glutathione (GSH) and its related synthase glutamate‐cysteine ligase catalytic subunit (GCLc) and glutamate‐cysteine ligase modifier subunit (GCLm) in ANIT‐treated group. In addition, the RNA and protein expression of Akt and nuclear factor‐E2‐related factor‐2 (Nrf2) were also activated by paeoniflorin in ANIT‐induced rats. These findings indicated that paeoniflorin protected ANIT‐induced cholestasis and increased GSH synthesis by activating Nrf2 through PI3K/Akt‐dependent pathway. Therefore, paeoniflorin might be a potential therapeutic agent for cholestasis. Copyright

Microcalorimetric investigation of the antibacterial activity of curcumin on Staphylococcus aureus coupled with multivariate analysis

The antibacterial effect of Curcumin on Staphylococcus aureus growth was evaluated by microcalorimetry. The heat flow power–time curves and nine quantitative parameters of the S. aureus growth were applied to investigate the inhibitory effect with Curcumin. By analyzing these curves and some quantitative parameters using multivariate analytical methods, similarity analysis and principal component analysis, the antibacterial activity of Curcumin on S. aureus could be accurately evaluated from the change of the two main parameters, the second exponential growth rate constant k2 and the maximum heat flow power Pm2. The main two thermal parameters played more important role in the evaluation: at low concentration (0–10.5xa0μgxa0mL−1), Curcumin hardly influence the growth of S. aureus, while at high concentration (10.5–43.4xa0μgxa0mL−1) it could notably inhibit the growth. All these illustrated that the antibacterial activity of Curcumin on S. aureus was enhanced with the increase of the concentration of this compound. This study might provide an useful method and idea accurately evaluate the antibacterial effects of Curcumin, which provides some useful methods for evaluate the nature antibacterial agents.

Microcalorimetric investigation of five Aconitum L. plants on the metabolic activity of mitochondria isolated from rat liver

The fact that mitochondria still have metabolic activity and can live for a week with appropriate nutrients after isolated from the rat liver provides us a useful means to investigate the metabolic process of mitochondria influenced by chemicals. Using the TAM air isothermal microcalorimeter, the HP-time curves of the metabolic activity of mitochondria were measured. The effects of five Aconitum L. plants, Radix Aconiti Lateralis Preparata (RALP), Radix Aconiti (RA), Radix Aconiti Kusnezoffii (RAK), Radix Aconiti Brachypodi (RAB), and Radix Aconiti Singularis (RAS) on the metabolic activity of mitochondria were investigated, respectively. The quantitative information, such as k, Q, Pmax, Tmax, Tlag, and Pav obtained from the HP-time curves, were accumulated for analysis. With the help of PCA method, Pav was selected as the standard for comparing the different effects of five Aconitum L. plants on metabolic activity of mitochondria. The potential sequence of efficiency was RASxa0>xa0RAKxa0>xa0RABxa0>xa0RAxa0>xa0RALP, and the EC50 were separately 1.43, 2.31, 1.10, 1.61, and 2.56xa0mgxa0mL−1. The results demonstrated that the five Aconitum L. plants had different bioeffects on the mitochondria metabolism. Meanwhile, this study indicated that microcalorimetry was a powerful tool to evaluate the drugs’ efficiency on living system, providing some useful references for the application of five Aconitum L. plants in practice.

Microcalorimetry and turbidimetry to investigate the anti-bacterial activities of five fractions from the leaves of Dracontomelon dao on P. aeruginosa

As a healthcare-associated pathogen with the resistance to anti-bacterial agent, P. aeruginosa has caused prevalent public burden and should not be ignored. Facing the realistic situation, developing novel anti-P. aeruginosa agents is urgent. Leaves of Dracontomelon dao were extensively used in southern China to treat various infectious diseases 1000xa0years ago. And in the study, the heat flow power–time (HFP–time) curves generated by P. aeruginosa which were disturbed by the five fractions (PE, CHCl3, EtOAc, n-BuOH and Vestiges fraction) from them were investigated through microcalorimetry, and then, some thermal kinetic parameters were obtained from the curves to characterize the metabolism of P. aeruginosa. The parameters were analyzed by principal component analysis (PCA), and the anti-P. aeruginosa activities of the five fractions were systematically compared and evaluated. The results showed that five fractions all expressed respective anti-P. aeruginosa effects in a dose-dependent manner and especially the performance of EtOAc fraction with half-inhibitory concentration (IC50) of 18.06xa0μgxa0mL−1. Simultaneously, the prospective performance of EtOAc fraction was confirmed by turbidimetry. Based on the promising anti-P. aeruginosa activities of EtOAc fraction, it could be developed as a novel anti-bacterial agent used in practice for treating certain infectious diseases.

Study on Drug Property Differences of Shexiang (Moschus) and Bingpian (Borneolum Synthcticum) Based on Analysis of Biothermodynamics

OBJECTIVEnTo study the drug property differences of Shexiang (Moschus) and Bingpian (Borneolum Synthcticum) with biothermodynamics, and to verify the objectivity and authenticity of the drug property.nnnMETHODSnThe growth-thermogram curves of Escherichia coli (E. coli) affected by Shexiang (Moschus) and Bingpian (Borneolum Synthcticum) at different concentrations were determined with microcalorimetry, and the power-time curves (thermogram curves) of E. coli metabolism and characteristic parameters, such as growth rate constant (k), maximum output power (P(m)), peak time (t(p)), total heat-output (Q(t)), etc. were analyzed with the principal component analysis (PCA) to find the close correlative parameters, so as to objectively reflect the drug property differences of Shexiang (Moschus) and Bingpian (Borneolum Synthcticum).nnnRESULTSnThe values of P2 in the second exponential growth phase increased with the increase of the concentrations of Shexiang (Moschus) and Bingpian (Borneolum Synthcticum), and the P2 of Shexiang (Moschus) was larger than that of Bingpian (Borneolum Synthcticum); Q2 increased with the increase of the concentrations of Shexiang (Moschus), but for Bingpian (Borneolum Synthcticum) it was opposite. It is indicated that they have different effects on P2 and Q2 of E. coli in the second exponential growth phase, and have differences in warm and cold natures.nnnCONCLUSIONnThe microcalorimetry can accurately and objectively appraise differences of the drug property of Shexiang (Moschus) and Bingpian (Borneolum Synthcticum) and verify the objectivity and authenticity of the drug property, so as to provide a new and useful method for studies of the drug property of Chinese drugs.

Microcalorimetry coupled with principal component analysis for investigating the anti-Staphylococcus aureus effects of different extracted fractions from Dracontomelon dao

With the prevalence resistance of Staphylococcus aureus to antibacterial agents, developing novel antibacterial agents is urgent. Recently, plant extracts have got more focus. In this study, the power-time curves produced by S. aureus under the action of the four extracted fractions (PE, CHCl3, EtOAc, and n-BuOH fractions) from the leaves of Dracontomelon dao were determined by microcalorimetry, and then some quantitative parameters, such as growth rate constant k, total heat output Qt, maximum heat-output power Pm, and the appearance time tm were obtained. By analyzing the parameters using principal component analysis, the anti-S. aureus effects of the four fractions were systematically evaluated and compared. Meanwhile, the total flavonoid contents in these fractions were analyzed. The results have evidenced that different fractions using various extraction solvents expressed various anti-S. aureus effects, and the inhibitory effects were presented in a flavonoid content-dependent manner. The EtOAc fraction with the highest total flavonoid content (41.86xa0%) expressed the strongest anti-S. aureus effect with half-inhibitory concentration (IC50) of 83.93xa0μgxa0mL−1, which might be applied as a novel antibacterial agent in practice for some infectious diseases. In addition, the microcalorimetric method should be strongly suggested in screening for novel antibacterial agents for fighting against pathogenic bacteria.