Radix Paeoniae Alba attenuates Radix Bupleuri-induced hepatotoxicity by modulating gut microbiota to alleviate the inhibition of saikosaponins on glutathione synthetase / 药物分析学报

Radix Bupleuri(RB)is commonly used to treat depression,but it can also lead to hepatotoxicity after long-term use.In many anti-depression prescriptions,RB is often used in combination with Radix Paeoniae Alba(RPA)as an herb pair.However,whether RPA can alleviate RB-induced hepatotoxicity remain unclear.In this work,the results confirmed that RB had a dose-dependent antidepressant effect,but the optimal antidepressant dose caused hepatotoxicity.Notably,RPA effectively reversed RB-induced hepatotoxicity.Afterward,the mechanism of RB-induced hepatotoxicity was confirmed.The results showed that saiko-saponin A and saikosaponin D could inhibit GSH synthase(GSS)activity in the liver,and further cause liver injury through oxidative stress and nuclear factor kappa B(NF-KB)/NOD-like receptor thermal protein domain associated protein 3(NLRP3)pathway.Furthermore,the mechanisms by which RPA attenuates RB-induced hepatotoxicity were investigated.The results demonstrated that RPA increased the abundance of intestinal bacteria with glycosidase activity,thereby promoting the conversion of saikosaponins to sai-kogenins in vivo.Different from saikosaponin A and saikosaponin D,which are directly combined with GSS as an inhibitor,their deglycosylation conversion products saikogenin F and saikogenin G exhibited no GSS binding activity.Based on this,RPA can alleviate the inhibitory effect of saikosaponins on GSS activity to reshape the liver redox balance and further reverse the RB-induced liver inflammatory response by the NF-κB/NLRP3 pathway.In conclusion,the present study suggests that promoting the conversion of saikosa-ponins by modulating gut microbiota to attenuate the inhibition of GSS is the potential mechanism by which RPA prevents RB-induced hepatotoxicity.

Effects of Kruppel-like factor 6 on HepG2 and the development of liver in zebrafish / 中国比较医学杂志

Objective To investigate the effects of Kruppel-like factor 6 (KLF6) on the apoptotic and migration ability of HepG2 cell, and the developmental role of KLF6 on zebrafish liver.Methods Constructed plasmid with shRNA-KLF6 was transfected in HepG2 and L-02.The impacts of loss of KLF6 on HepG2 was investigated by Western bolt, apoptosis analyses, cell cycle detection and scratch experiment;KLF6 morpholino oligonucleotides was microinjected into the Tg(lfabp:eGFP) transgenic zebrafish embryos.The morphant phenotype of the liver was imaged and the protein expression of KLF6 after knockdown of KLF6 was analyzed by Immunofluorescence staining.Results The expression of KLF6 in L-02 was significantly higher than in HepG2.After knockout of KLF6, KLF6 protein expression and apoptosis were significantly reduced.In addition, the cell cycle mainly stated in S phase and the migration ability of HepG2 was enhanced.After klf6 knockdown in transgenic zebrafish larvae, the development of zebrafish liver was delayed and KLF6 expression was obviously decreased in the liver.Conclusions The reduction of KLF6 expression increased the proliferation and migration ability, and reduced the apoptosis of HepG2.Loss of KLF6 affects the development of zebrafish liver, which may open a possibility to use zebrafish as a liver cancer model and for anti-liver cancer drug screening.

Simultaneous determination of six constituents in Liuhe Dingzhong Pills by HPLC / 中成药

AIM To establish an HPLC method for the simultaneous content determination of narirutin,naringin,hesperidin,neohesperidin,honokiol and magnolol in Liuhe Dingzhong Pills (Aurantii Fructus,Citri reticulatae Pericarpium,Magnoliae officinalis Cortex,etc.).METHODS The analysis of 75％ methanol extract of this drug was performed on a 30 ℃ thermostatic Dikma Spursil C18 column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of acetonitrile-water flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelengths were set at 283 nm and 294 nm.RESULTS Six constituents showed good liner relationships within their own ranges (r≥0.999 3),whose average recoveries were 92.6％-103.7％ with the RSDs of 0.89％-2.87％.The contents of naringin and neohesperidin in three batches of samples showed obvious differences.CONCLUSION We should pay attention to the unstable quality of Liuhe Dingzhong Pills due to Aurantii Fructus.

An exploration in the action targets for antidepressant bioactive components of Xiaoyaosan based on network pharmacology / 药学学报

The present study aims to predict the action targets of antidepressant active ingredients of Xiaoyaosan to understand the "multi-components, multi-targets and multi-pathways" mechanism. Using network pharmacology, the reported antidepressant active ingredients in Xiaoyaosan (saikosaponin A, saikosaponin C, saikosaponin D, ferulic acid, Z-ligustilide, atractylenolide I, atractylenolide II, atractylenolide III, paeoniflorin, albiflorin, liquiritin, glycyrrhizic acid and pachymic acid), were used to predict the targets of main active ingredients of Xiaoyaosan according to reversed pharmacophore matching method. The prediction was made via screening of the antidepressive drug targets approved by FDA in the DrugBank database and annotating the information of targets with the aid of MAS 3.0 biological molecular function software. The Cytoscape software was used to construct the Xiaoyaosan ingredients-targets-pathways network. The network analysis indicates that the active ingredients in Xiaoyaosan involve 25 targets in the energy metabolism-immune-signal transmutation relevant biological processes. The antidepressant effect of Xiaoyaosan reflects the features of traditional Chinese medicine in multi-components, multi-targets and multi-pathways. This research provides a scientific basis for elucidation of the antidepressant pharmacological mechanism of Xiaoyaosan.

Molecular identification of one Uncaria plant / 中国中药杂志

<p><b>OBJECTIVE</b>In order to identify a species of Uncaria, molecular phylogenetic analysis was carried out by using the rDNA ITS sequence as molecular marker.</p><p><b>METHOD</b>Total DNA was extracted from the plant with modified CTAB method and thereby rDNA ITS regions were amplified with universally conserved primer. The rDNA ITS amplicon was characterized by cloning, sequencing, blasting in GenBank and phylogenetic analyses using PAUP by maximum parsimony (MP) criteria.</p><p><b>RESULT</b>The rDNA ITS entire sequence of this species of Uncaria was 719 bp. The sequence is related to the U. sinensis available in GenBank and the similarity reaches 99.7%.</p><p><b>CONCLUSION</b>Based on molecular biology methods of rDNA ITS region analysis, molecular identification is available in accurate classification on this species of Uncaria.</p>

Simultaneous Determination of Peoniflorin, Campanulin and Ferulic Acid in Kangmin Granules by HPLC / 中国药房

OBJECTIVE: To establish an HPLC method for simultaneous determination of peoniflorin,calycosin and ferulic acid in Kangmin granules.METHODS: The separation was performed on a Kromasil C18 column (250 mm?4.6 mm,5 ?m),and the mobile phase consisted of methanol-acetonitrile-0.4% phosphonic acid (20 ∶ 10 ∶ 70) with flow rate of 1.0 mL?min-1.The detection was performed at wavelength of 254 nm.Column temperature was room temperature and the injection volume was 10 ?L.RESULTS: The linear ranges of peoniflorin,calycosin and ferulic acid were 180～3 600 ?g?mL-1(r=0.999 2),4.6～92.0 ?g?mL-1(r=0.999 1) and 8.0～160.0 ?g?mL-1(r=0.999 4),respectively.The average recoveries were 98.3%(RSD=1.8%,n=9),99.0% (RSD=2.0%,n=9) and 100.5%(RSD=1.1%,n=9),respectively.CONCLUSION: The method is simple,accurate,sensitive and reproducible for the quality control of Kangmin granules.

Proliferations of T cell lines towards neural myelin sheath components in multiple sclerosis induced by neural myelin sheath and delipidated neural myelin sheath / 中国组织工程研究

BACKGROUND: Multiple sclerosis(MS) is a chronic autoimmune disease induced by the interaction between genetic and environmental factors. Its pathogen and the mechanism of the relapse and remission m the course of the disease are still unknown. Most of the MS research centers are looking for the pathogenic polypeptide epitope in proteolipid protein(PLP), myelin sheath basic protein (MBP) and oligodendrocyte glycoprotein (MOG) OBJECTIVE: To compare the proliferation of T cell lines(TCL) in MS induced by myelin sheath and delipidated myelin sheath towards 11 components of myelin sheath to mainly search the possible pathogenic polypeptide epitope in PLP, and investigate the possible effects of abnormal dcgrease in myelin sheath.DESIGN: A case-controlled trial.SETTING: Department of neurology in a hospital of a university.PARTICIPANTS: Mononuclear cells(MNC) of 16 MS cases(clinical relapsing-remitting type, patients did not receive any immunosuppresant for at least 3 months when their peripheral blood samples were taken) and 12 HLA-DR15 healthy volunteers were furnished by Dr. Trotter JL of MS Research Center of Washington University from the cell database.INTERVENTIONS: MS-TCL and normal TCL were induced twice by stimulation with myelin sheath and delipidated myelin sheath in vitro by cell culture in vitro. TCL proliferation was tested by 11 antigens including PLP,MBP, M87-106, P30-49, P40-60, P89-106, P95-117, P117-137,P139-151, P178-191, and P185-206.MAIN OUTCOME MEASURES: Difference of scintillation counting in every minute of every well, and the stimulative index of each well were calculated, and the mean wells with positive proliferation of TCL towards each antigen were confirmed as well.RESULTS: The general specific proliferation towards myelin sheath antigens was bigger in MS group than control group 5.49 ±5.31 to 3.10 ± 3. 17, and delipidated myelin sheath-induced TCL was bigger than myelin sheath-induced one 5. 49 ± 5.31 to 3.41 ± 4. 83 . Delipidated myelin sheath significantly changed the immune responses of MS group,especially the changes of responses towards P30-49, P40-60, P89-106,P117-137, P139-161, and P185-206 were significant compared with that the control group only responded to two polypeptides, which indicated that the antigen epitope of MBP, PLP, M87-106, P95-117, P40-60, and P185-206 might have significance in the triggering of MS autoimmune responses.CONCLUSION: TCL induced by MS myelin sheath has different proliferation towards antigen components of myelin sheath from control group. Delipidated myelin sheath significantly increases TCL proliferation in MS group, which suggests that if MS patients developed abnormal degrease in myelin sheath, TCL would produce autoimmune response towards self-myelin sheath, MBP, PLP and its polypeptide segments all can trigger MS or aggravate the state of the illness. Our finding supports the hypothesis of MS autoimmune pathogenic mechanism.

Determination of Forsythin in Baohe Oral Liquid by HPLC / 中药新药与临床药理

Objective To establish a method for the determination of forsy thin in Baohe Oral Liquid by HPLC.Methods The determination was carried out on a Diamonsil C18 column,with mobile phase of acetonitrile-water(29∶71) at room temperature.The flow rate was 1.0mL?min-1 and the detective wavelength at 277 nm.Results The calibration curves was linear in the range of 0.068～ 0.340 ? g(r=0.9996).The average recovery rate of tested sample was 99.7 %(RSD=2.1 %).Conclusion The method was specific,accurate and precise.It can be used for the quality control of Baohe Oral Liquid.