Network Pharmacology Combined with Bioinformatics Analysis to Texplore the Potential Mechanism of Phellodendri Chinensis Cortex Against Bladder Cancer.

The pharmacological mechanism of Phellodendri Chinensis cortex (PCC) against diseases, especially bladder cancer (BC), has never been reported systematically. This study was designed to explore potential mechanism of PCC in treatment of BC. First, we used network pharmacology to discover the potential mechanism of Phellodendri Chinensis cortex and phellodendrine against bladder cancer. Then, we used bioinformatics analysis to verify the correlation between gene expression analysis, survival analysis and common targets. Finally, molecular docking was used to calculate the binding energies of phellodendrine and common targets.A total of 264 targets for PCC were predicted, and 391 BC-related targets were obtained from 4 databases. There were 54 potential targets, 315 biological processes, and 120 signaling pathways involved for PCC against BC. The CDKN2A expression increased and the ESR1, JUN, IL6, AR, and PTGS2 levels decreased in BC according to Gene Expression Profiling Interactive Analysis version 2. The high expression of JUN, MYC, EGFR, and EGF and low expression of VEGFA and PPARG were associated with short overall survival (OS). The high expression of AKT1, EGFR, and EGF and low expression of IL1ß were associated with poor disease-free survival (DFS). The search of the intersection of phellodendrine and BC targets yielded 11 common targets, 50 biological processes, and 13 signaling pathways involved. High AURKA and FASN and low ESR1, JUN, ABCB1, and PTGS1 were expressed in BC. The high expression of FASN, ABCC1, PTGS1, JUN, and PIK3CA was associated with short OS, the high expression of PIK3CA and ABCC1 was associated with poor DFS prognosis. Phellodendrine showed a better binding affinity for PTGS2 protein with a docking score of -7.183 and a MM-GBSA result of -46.47 kcal/mol. This study revealed potential mechanism of PCC and phellodendrine against BC through network pharmacology and bioinformatics.

Screening and Mechanism Study of Three Antagonistic Drugs, Oxysophoridine, Rutin, and Phellodendrine, against Zearalenone-Induced Reproductive Toxicity in Ovine Oocytes.

Zearalenone (ZEN) is a common fungal toxin with reproductive toxicity in various grains. It poses a serious threat to ovine and other animal husbandry industries, as well as human reproductive health. Therefore, investigating the mechanism of toxicity and screening antagonistic drugs are of great importance. In this study, based on the natural compound library and previous Smart-seq2 results, antioxidant and anti-apoptotic drugs were selected for screening as potential antagonistic drugs. Three natural plant compounds (oxysophoridine, rutin, and phellodendrine) were screened for their ability to counteract the reproductive toxicity of ZEN on ovine oocytes in vitro using quantitative polymerase chain reaction (qPCR) and reactive oxygen species detection. The compounds exhibited varying pharmacological effects, notably impacting the expression of antioxidant (GPX, SOD1, and SOD2), autophagic (ATG3, ULK2, and LC3), and apoptotic (CAS3, CAS8, and CAS9) genes. Oxysophoridine promoted GPX, SOD1, ULK2, and LC3 expression, while inhibiting CAS3 and CAS8 expression. Rutin promoted SOD2 and ATG3 expression, and inhibited CAS3 and CAS9 expression. Phellodendrine promoted SOD2 and ATG3 expression, and inhibited CAS9 expression. However, all compounds promoted the expression of genes related to cell cycle, spindle checkpoint, oocyte maturation, and cumulus expansion factors. Although the three drugs had different regulatory mechanisms in enhancing antioxidant capacity, enhancing autophagy, and inhibiting cell apoptosis, they all maintained a stable intracellular environment and a normal cell cycle, promoted oocyte maturation and release of cumulus expansion factors, and, ultimately, counteracted ZEN reproductive toxicity to promote the in vitro maturation of ovine oocytes. This study identified three drugs that antagonize the reproductive toxicity of ZEN on ovine oocytes, and compared their mechanisms of action, providing data support and a theoretical basis for their subsequent application in the ovine breeding industry, reducing losses in the breeding industry, screening of ZEN reproductive toxicity antagonists and various toxin antagonists, improving the study of ZEN reproductive toxicity mechanisms, and even protection of human reproductive health.

Inhibitory effect of phellodendrine on C48/80-induced allergic reaction in vitro and in vivo.

The incidence of allergic reactions has risen steadily in recent years, prompting growing interest in the identification of efficacious and safe natural compounds that can prevent or treat allergic diseases. Phellodendron amurense Rupr. has long been applied as a treatment for allergic diseases, whose primary component is phellodendrine. However, the efficacy of phellodendrine as a treatment for allergic diseases remains to be assessed. Mast cells are the primary effectors of allergic reactions, which are not only activated by IgE-dependent pathway, but also by IgE-independent pathways via human MRGPRX2, rat counterpart MRGPRB3. As such, this study explored the effect and mechanism of phellodendrine through this family receptors in treating allergic diseases in vitro and in vivo. These analyses revealed that phellodendrine administration was sufficient to protect against C48/80-induced foot swelling and Evans blue exudation in mice, and suppressed C48/80-induced RBL-2H3 rat basophilic leukemia cells degranulation, and ß-HEX, HIS, IL-4, and TNF-α release. Moreover, phellodendrine could reduce the mRNA expression of MRGPRB3 and responsiveness of MRGPRX2 by altering its structure. It was able to decrease Ca2+ levels, phosphorylation levels of CaMK, PLCß1, PKC, ERK, JNK, p38, and p65, and inhibit the degradation of IκB-α. These analyses indicate that berberine inhibits the activation of PLC and downregulates the release of Ca2+ in the endoplasmic reticulum by altering the conformation of MRGPRB3/MRGPRX2 protein, thereby inhibiting the activation of PKC and subsequently inhibiting downstream MAPK and NF-κB signaling, ultimately suppressing allergic reactions. There may thus be further value in studies focused on developing phellodendrine as a novel anti-allergic drug.

Biological Importance of Phellodendrine in Traditional and Modern Medicines: An Update on Therapeutic Potential in Medicine.

BACKGROUND: Herbal medicines have been used for the preparation of numerous pharmaceutical products for the treatment of human disorders. Plant-derived products have been used in medicine, nutraceuticals, perfumery, beverages, and cosmetics industries for different purposes. Herbal medicines are mainly derived from different parts of plant materials. Phellodendron bark has been widely known as one of the fundamental herbs of traditional Chinese medicine. Phellodendron bark contains phellodendrine as a main active phytochemical. Phellodendrine ((7S,13aS)-3,10-dimethoxy-7-methyl-6,8,13,13atetrahydro-5H-isoquinolino[2,1-b]isoquinolin-7-ium-2,11-diol), is a quaternary ammonium alkaloid. METHODS: This present study aimed to investigate the biological potential and therapeutic effectiveness of phellodendrine in medicine through scientific data analysis of different research works on phellodendrine. The therapeutic value of phellodendrine was analyzed in the present work through scientific data available in Google, Google Scholar, ScienceDirect, and PubMed. All the scientific data on phellodendrine were collected from these databases using the terms herbal drugs and phellodendrine. Pharmacological and analytical data of phellodendrine were analyzed in the present work in order to know the medicinal importance of phellodendrine. RESULTS: Scientific data analysis of phellodendrine in the present work signified the biological importance of phellodendrine in medicine. Phellodendrine has numerous beneficial aspects in medicine due to its potential benefits in ulcerative colitis, inflammation, pancreatic cancer, nephritis, immune response, acetylcholinesterase activity, psoriasis, arthritis, atopic dermatitis, and oxidative stress. However, it also has significant effects on eicosanoid generation, neuraminidase-1, inflammasome generation, cytochrome p450, taste receptors, and hepatic gluconeogenesis. Furthermore, scientific data has indicated the presence of phellodendrine in different natural sources, including Phellodendri cortex. Analytical data on phellodendrines has signified their importance in the isolation and separation of pure phytochemicals in medicine. Pharmacokinetic parameters have highlighted the tissue distribution of phellodendrine in different tissue of human beings and higher animals. CONCLUSION: In the present work, scientific data analysis has indicated the biological importance, pharmacological activities, and analytical aspects of phellodendrine in medicine.

Utilizing network pharmacology and experimental validation to investigate the underlying mechanism of phellodendrine on inflammation.

Background: Phellodendrine, one of the characteristic and important active components of Cortex phellodendri, has been proven to show anti-inflammatory effects. However, the underlying mechanism of phellodendrine on inflammation remains largely unclear. Aim of the study: In this study, network pharmacology and experimental validation were used to explore the underlying mechanism of phellodendrine on inflammation. Materials and Methods: PubChem and SwissADME database were used to evaluate the drug-likeness and other characteristics of phellodendrine. The targets of phellodendrine for the treatment of inflammation were analyzed with multiple databases. Other extensive analyses including protein-protein interaction, Gene Ontology, and Kyoto Encyclopedia of Genes and Genomes pathway enrichment were accomplished with the STRING database, Cytoscape software, and DAVID database. Moreover, the effect of phellodendrine on anti-inflammation was proven in RAW264.7. Results: The network pharmacology results indicated that phellodendrine had drug potential. Phellodendrine acted directly on 12 targets, including PTGS1, PTGS2, HTR1A, and PIK3CA, and then regulated cAMP, estrogen, TNF, serotonergic synapse, and other signaling pathways to exert anti-inflammatory effects. The experimental results showed that phellodendrine reduced the levels of IL-6 compared with the LPS group in 24 h and changed the mRNA expression of PTGS1, PTGS2, HSP90ab1, AKT1, HTR1A, PI3CA, and F10. Conclusion: Our research preliminarily uncovered the therapeutic mechanisms of phellodendrine on inflammation with multiple targets and pathways. Phellodendrine may be a potential treatment for inflammation-related diseases related to the cAMP and TNF signaling pathways.

Phellodendrine promotes autophagy by regulating the AMPK/mTOR pathway and treats ulcerative colitis.

To investigate the therapeutic effects of phellodendrine in ulcerative colitis (UC) through the AMPK/mTOR pathway. Volunteers were recruited to observe the therapeutic effects of Compound Cortex Phellodendri Liquid (Huangbai liniment). The main components of Compound Cortex Phellodendri Liquid were analysed via network pharmacology. The target of phellodendrine was further analysed. Caco-2 cells were cultured, and H2 O2 was used to stimulate in vitro cell model. Expression levels of LC3, AMPK, p-AMPK, mTOR and p-mTOR were detected via Western blotting and through immunofluorescence experiments. The therapeutic effects of phellodendrine were analysed via expression spectrum chip sequencing. The sequencing of intestinal flora further elucidated the therapeutic effects of phellodendrine. Compared with the control group, Compound Cortex Phellodendri Liquid could substantially improve the healing of intestinal mucosa. Network pharmacology analysis revealed that phellodendrine is the main component of Compound Cortex Phellodendri Liquid. Moreover, this alkaloid targets the AMPK signalling pathway. Results of animal experiments showed that phellodendrine could reduce the intestinal damage of UC compared with the model group. Findings of cell experiments indicated that phellodendrine treatment could activate the p-AMPK /mTOR signalling pathway, as well as autophagy. Expression spectrum chip sequencing showed that treatment with phellodendrine could promote mucosal healing and reduce inflammatory responses. Results of intestinal flora detection demonstrated that treatment with phellodendrine could increase the abundance of flora and the content of beneficial bacteria. Phellodendrine may promote autophagy by regulating the AMPK-mTOR signalling pathway, thereby reducing intestinal injury due to UC.

In vitro inhibitory effects of phellodendrine on human liver cytochrome P450 enzymes.

1. Phellodendrine possesses numerous pharmacological activities. However, whether phellodendrine affects the activity of human liver cytochrome P450 (CYP) enzymes remains unclear.2. In this study, the inhibitory effects of phellodendrine on eight human liver CYP isoforms (i.e. 1A2, 3A4, 2A6, 2E1, 2D6, 2C9, 2C19 and 2C8) were investigated in vitro using human liver microsomes (HLMs).3. The results showed that phellodendrine inhibited the activity of CYP1A2, 3A4 and 2C9, with IC50 values of 20.56, 14.98 and 16.30 µM, respectively, but that other CYP isoforms were not affected. Enzyme kinetic studies showed that phellodendrine was not only a non-competitive inhibitor of CYP3A4, but also a competitive inhibitor of CYP1A2 and 2C9, with Ki values of 7.15, 10.52 and 7.98 µM, respectively. In addition, phellodendrine is a time-dependent inhibitor for CYP3A4 with Kinact/KI value of 0.046/11.57 µM-1 min-1.4. The in vitro studies of phellodendrine with CYP isoforms indicate that phellodendrine could inhibit the activities of CYP1A2, 3A4 and 2C9. Further clinical studies are needed to evaluate the significance of this interaction.

Phellodendrine chloride suppresses proliferation of KRAS mutated pancreatic cancer cells through inhibition of nutrients uptake via macropinocytosis.

Despite the massive efforts to develop the treatment of pancreatic cancers, no effective application exhibits satisfactory clinical outcome. Macropinocytosis plays a critical role for continuous proliferation of pancreatic ductal adenocarcinoma (PDAC). In this study, we generated a screening method and identified phellodendrine chloride (PC) as a potential macropinocytosis inhibitor. PC significantly inhibited the viability of KRAS mutant pancreatic cancer cells (PANC-1 and MiaPaCa-2) in a dose-dependent manner; however, it did not affect the wild type KRAS pancreatic cancer cells (BxPC-3). Further experiments indicated that PC reduced the growth of PANC-1 cells through inhibition of macropinocytosis and diminishing the intracellular glutamine level. Disruption of glutamine metabolism led to enhance the reactive oxygen species level and induce mitochondrial membrane potential depolarization in PANC-1 cells. PC treatment caused increased Bax and decreased Bcl-2 expression, along with the activation of cleaved caspase-3, 7, 9 and cleaved-PARP, thus induced mitochondrial apoptosis. Moreover, PC inhibited macropinocytosis in vivo and effectively reduced the growth of PANC-1 xenograft tumors. All together, we demonstrated that inhibition of macropinocytosis might be an effective strategy to treat pancreatic cancers. Thus, PC could be a potential compound with improved therapeutic efficacy in patients with pancreatic cancers.

Protective effect of phellodendrine on bacterial vaginosis mice / 中草药

Objective To investigate the impact of phellodendrine on treating bacterial vaginosis (BV) mice. Methods BV model mice were induced by vaginal injecting with EPEC and MRSA. After the mice in control and model groups being administrated with phellodendrine (10, 20, and 40 mg/kg) for 7 d, the mice were sacrificed under deep anesthesia. The mice vaginal pathology changes were observed using HE staining method. IL-1β, IL-6, IL-8, and TNF-α secretion in vagina and PGE2, PGF2α levels in uterus were detected using ELISA assay. Uterine MMP-9, TLR4, and NF-κB expression levels were measured using western blotting. Results Vaginal pathology changes were improved by phellodendrine. Specifically, phellodendrine could reduce IL-1β, IL-6, IL-8, and TNF-α secretion in vagina, it could also reduce PGE2 and PGF2α content, and inhibit MMP-9, TLR4, and NF-κB expression levels in uterus tissue. Conclusion Phellodendrine could effectively treat BV and reduce the rise of pre-term birth by simultaneously inhibiting endovaginal inflammatory response and regulating intrauterine TLR4/NF-κB signal pathway in BV mice.

Phytochemical Quantification and the In Vitro Acetylcholinesterase Inhibitory Activity of Phellodendron chinense and Its Components.

The dried bark of Phellodendron chinense has been used as a traditional herbal medicine to remove damp heat, relieve consumptive fever, and cure dysentery and diarrhea. In the present study, we performed quantitative analyses of the two components of P. chinense, phellodendrine and berberine, using high-performance liquid chromatography. A 70% ethanol extract of P. chinense was prepared and the two components were separated on a C-18 analytical column using a gradient solvent system of acetonitrile and 0.1% (v/v) aqueous trifluoroacetic acid. The ultraviolet wavelength used for detection was 200 nm for phellodendrine and 226 nm for berberine. The analytical method established here showed high linearity (correlation coefficient, ≥0.9991). The amount of phellodendrine and berberine used was 22.255 ± 0.123 mg/g and 269.651 ± 1.257 mg/g, respectively. Moreover, we performed an in vitro acetylcholinesterase (AChE) activity assay and an amyloid-ß aggregation test to examine the biological properties of phellodendrine and berberine as therapeutic drugs for Alzheimer's disease. Phellodendrine and berberine inhibited AChE activity in a dose-dependent manner (IC50 = 36.51 and 0.44 µM, respectively). In contrast, neither phellodendrine nor berberine had an effect on amyloid-ß aggregation. The P. chinense extract and phellodendrine, but not berberine, exhibited antioxidant activity by increasing radical scavenging activity. Moreover, P. chinense demonstrated a neuroprotective effect in hydrogen peroxide-treated HT22 hippocampal cells. Overall, our findings suggest that P. chinense has potential as an anti-Alzheimer's agent via the suppression of the enzymatic activity of acetylcholinesterase and the stimulation of antioxidant activity.

Simultaneous Determination of 4 Components in Shenbai Shuxin Granules by HPLC / 中国药房

OBJECTIVE:To establish a method for simultaneous determination of sodium danshensu,protocatechuic aldehyde,salvianolic acid B and phellodendrine hydrochloride in Shenbai shuxin granules.METHODS:HPLC method was adopted.The separation was performed on Waters sunfire-C18 column with mobile phase consisted of acetonitrile-0.05 ％ trifluoroacetic acid (gradient elution) at the flow rate of 1.0 mL/min.The detection wavelength was set at 288 nm,and the column temperature was 30 ℃.The sample size was 5 μL.RESULTS:The linear ranges of sodium danshensu,protocatechuic aldehyde,salvianolic acid B and phellodendrine hydrochloride were 0.040 01-1.600 46 μg(r=0.999 9),0.013 84-0.553 7 μg(r=0.999 9),0.049 32-1.972 94 μg(r=0.999 6),0.014 46-0.578 6 μg(r=0.999 8).The limits of quantitation were 7.68,2.66,4.74,1.38 ng,and the limits of detection were 1.92,0.66,2.36,0.69 ng,respectively.RSDs of precision,stability and reproducibility tests were all lower than 2％.The recoveries were 98.846％-100.762％ (RSD=0.77％,n=6),96.632％-99.463％ (RSD=0.98％,n=6),98.541％-100.432％ (RSD=0.82 ％,n =6),98.607 ％-101.521％ (RSD =1.11％,n =6),respectively.CONCLUSIONS:The method is simple,accurate and precise.It can be used for 4 components in Shenbai shuxin granules.

Simultaneous Determination of Five Components in Ermiao Pills by UHPLC with Dual-Wavelength / 中国药学杂志

OBJECTIVE: To develop an ultra-high performance liquid chromatography(UHPLC) method for simultaneous determination of berberine hydrochloride, phellodendrine hydrochloride, palmatine hydrochloride, magnoflorine, and rhizoma atractylodis in Ermiao pills. METHODS: The UHPLC analyses were performed on a Waters BEH C18 column (2.1 mm×100 mm, 1.7 μm)eluted with acetonitrile and 0.1% phosphoric acid(each 100 mL containing 0.1 g sodium dodecyl sulfate). The flow rate was 0.3 mL·min-1, the detection wavelength was set at 280 and 340 nm, and the column temperature was set at 30℃. RESULTS: The calibration curves of the five compositions had good linear relationship in the ranges of the tested concentrations. The precision, stability and repeatability complied with the requirements of methodology. The recoveries were between 96.8%-99.3% respectively. The RSDs were below 2.8%. CONCLUSION: The developed method is simple, sensitive, rapid and accurate. This work provides helpful information for the comprehensive quality evaluation of Ermiao pills.

Determination of Four Kinds of Ingredients in Transparent Absorbent Fluid of Different Size of Huoxue Powder by Ultra Performance Liquid Chromatography Tandem Mass Spectrometry / 医药导报

Objective To optimize the particle size of Huoxue powder by contents comparison of emodin,phellodendrine,berberine and jatrorrhizine before and after permeabilized skin absorption of Huoxue powder in different particle size of 0.150,0.075,0.048,0.038 mm.Methods The contents of emodin,phellodendrine,berberine and jatrorrhizine in transparent absorbent fluid of Huoxue power in different particle size within 24 hours were measured by ultra performance liquid chromatography tandem mass spectrometry,and optimize its particle size by contents comparison of the effective components.Results The contents of the active components in Huoxue power with the particle size of 0.075 mm were high before and after percutaneous absorption.Conclusion Particle size of 0.075 mm is best for Huoxue powder.

Content Determination of Phellodendrine in Leshu Lotion by SPE-HPLC / 中国中医药信息杂志

Objective To establish the method of thin layer identification of Phellodendri Chinensis Cortex and solid phase extraction - high performance liquid chromatography (SPE-HPLC) determination of the content of phellodendrine in Leshu Lotion. Methods Thin layer chromatography (TLC) was used for qualitative identification of Phellodendri Chinensis Cortex in Leshu Lotion and the Bond Elutplexa PCX strong cation-exchange solid phase was used for extraction small column purification. Platisil-NH2 column was as chromatographic column; column temperature was 30 ℃; triethylamine pH 3.82 (phosphoric acid), tetrahydrofuran, acetonitrile was 18:12:70, isocratic elution; flow rate was 1.0 mL/min; detection wavelength was 286 nm. Results TLC could obviously identify active ingredients of phellodendrine in Phellodendri Chinensis Cortex. The phellodendrine had good linear relationship between concentration and peak area within the scope of 1.22–12.2 μg (r=0.9999); the average recovery rate of phellodendrine was 101.18%, RSD was 1.71%. Conclusion This method is simple to operate, with accuracy and repeatability, and can be used for quality control of Leshu Lotion.

Pharmacokinetic studies of phellodendrine in rat plasma and tissues after intravenous administration using ultra-high performance liquid chromatography-tandem mass spectrometry.

Phellodendrine, a quaternary ammonium alkaloid extracted from the dried bark of Phellodendrom chinensis Schneid and Phellodendrom amurense Rupr, has the effect of suppressing cellular immune response, reducing blood pressure and antinephritis. However, few investigations have been conducted for the pharmacokinetic study of phellodendrine. Thus, a rapid, simple and reliable ultra-high performance liquid chromatography-tandem quadrupole mass spectrometry (UHPLC-QQQ MS/MS) method has been established for quantification of phellodendrine in rat plasma and tissues by using magnoflorine as internal standard. The chromatographic separation was achieved on an Agilent ZORBAX SB-C18 column (4.6mm×50mm, 1.8µm) by gradient elution using 0.1% aqueous formic acid (A) and methanol (B). Triple quadrupole mass detection with multiple reaction monitoring mode was used to monitor the ion transitions, at m/z 342.20→192.20 for phellodendrine and m/z 342.20→58.20 for internal standard, respectively. The developed method was fully validated and successfully applied to the pharmacokinetics and tissue distribution study of phellodendrine after intravenous administration. The lower limits of quantification were 0.5ng/mL for plasma samples, 2.5ng/g for brain and 1ng/g for other tested tissues. Precisions and accuracy values were within the Food and Drug Administration acceptance criteria, the recovery and matrix effects were between 87.8-113.5%. The area under the curve (AUC0-t) ranged from 15.58 to 57.41mg/L min and Cmax were between 1.63-4.93mg/L. The results showed that phellodendrine was eliminated in 120min in plasma and most of tissues and the highest concentrations of phellodendrine were found in the kidney. This study may provide a basis for the further study of phellodendrine.

The defensive effect of phellodendrine against AAPH-induced oxidative stress through regulating the AKT/NF-κB pathway in zebrafish embryos.

AIMS: This study is to investigate the effect of phellodendrine (PHE) against AAPH-induced oxidative stress and find out the biological mechanism of PHE by using the zebrafish embryo model. MAIN METHODS: After treatments by AAPH or PHE, the mortality and heartbeat of zebrafish embryos were recorded and the production of reactive oxygen species (ROS), lipid-peroxidation and the rate of cell death were detected by fluorescence spectrophotometry respectively. Whereafter, the pathways of PHE against AAPH-induced oxidative stress were screened by inhibitors to explore its biological mechanism. The related genes and proteins expressions were analyzed by real-time quantitative reverse-transcription polymerase-chain-reaction (qRT-PCR) and western blotting. KEY FINDINGS: The PHE obviously improved the decreased survival rate and abnormally elevated heart-beating rate of zebrafish embryos caused by AAPH. Especially 200µg/mL of PHE make the survival rate increased to 90.26±1.40% at 72hfp and the heartbeat back to normal. Besides, AAPH caused a significant increase in the production of reactive oxygen species (ROS), lipid-peroxidation and cell death rate, all of which could be decreased after PHE treatment dose-dependently. And PHE exerted the protective activity against AAPH-induced oxidative stress through down-regulating AKT phosphorylation and NF-kB3 expression, which associate with modulation of IKK phosphorylation in zebrafish embryos. SIGNIFICANCE: The PHE showed a good antioxidant effect in vivo, and the mechanism has been stated that the PHE can down-regulating AKT, IKK, NF-kB phosphorylation and COX-2 expression induced by AAPH. Moreover, the PHE also ameliorated the ROS-mediated inflammatory response.

Study on the Quality Standard of Huoxuesan / 中国药房

OBJECTIVE:To establish the quality standard of Huoxuesan. METHODS:Microscopic identification method was adopted for the qualitative identification of Rhizoma pinelliae,Eupolyphaga seu,Phellodendron amurense and Fructus kochiae in the preparation. HPLC was adopted for the contents determination of phellodendrine,polydatin,jatrorrhizine,berberine and emo-din:the column was ZORBAX Eclipse Plus C18 with methanol-0.1% phosphoric acid(gradient elution)at a flow rate of 0.4 ml/min,the detection wavelength was 240 nm,column temperature was 30 ℃,and the injection volume was 5 μl. RESULTS:Micro-scopic identification map of R. pinelliae,E. seu,P. amurense and F. kochiae was clear. The linear range was 0.159-5.073 μg(r=0.997 4)for phellodendrine、0.149-4.761 μg(r=0.999 9)for polydatin、0.239-7.649 μg(r=0.995 5)for jatrorrhizine、0.165-5.268 μg (r=0.997 2)for berberine、0.012-0.382 μg(r=0.999 9)for emodin;RSDs of precision,stability and reproducibility tests were low-er than 3.0%;recoveries were 98.9%-104.1%(RSD=1.9%,n=6),96.1%-101.7%(RSD=2.5%,n=6),99.6%-105.1%(RSD=2.6%,n=6),90.3%-98.2%(RSD=2.9%,n=6)and 92.9%-96.4%(RSD=2.0%,n=6)respectively. CONCLUSIONS:The es-tablished standard can be used for the quality control of Huoxuesan.

Study on Quality Standards forAnshu Liquid / 中国中医药信息杂志

Objective To establish quality standards forAnshu Liquid.Methods Phellodendri Chinensis Cortex and Kochiae Fructus were qualitatively identified by TLC method. The contents of berberine hydrochloride and phellodendrine hydrochloride in the preparation were determined by HPLC method. The chromatographic column was ZORBAX SB-C18 (4.6 mm×250 mm, 5μm); the mobile phase was acetonitrile-0.1% phosphoric acid (0.5% diethylamine) gradient elution; the flow rate was 1.0 mL/min; the detection wavelength was 284 nm.Results Berberine hydrochloride and phellodendrine hydrochloride in Phellodendri Chinensis Cortex and saponin Ic in Kochiae Fructus were detected by TLC method. The linear relationship of berberine hydrochloride in the preparation was Y=23.109X?30.548,r2=0.999 8, RSD=2.97%, showing that the linear range of berberine hydrochloride was 0.050 5– 2.525 0 μg. The linear relationship of phellodendrine hydrochloride in the preparation wasY=10.038X?2.446 6, r2=0.999 9, RSD=1.24%, showing that the linear range of phellodendrine hydrochloride was 0.050 0–2.500 0 μg. Conclusion The method is accurate, rapid, stable and reliable, with good reproducibility, which can be used for the quality control and evaluation ofAnshu Liquid.

HPLC fingerprint of analgesic ingredients in cangbaiqutong capsules / 国际药学研究杂志

Objective To establish fingerprint of analgesic ingredients in Cangbaiqutong capsules by HPLC, and control different production batches of Cangbaiqutong capsules’ quality. Methods Waters XSELECT CSH-C18 (4.6 mmxl50 mm, 5.0μm) chromatographic column was used and eluted with acetonitrile -0.1% phosphoric acid solution gradiently in the flow rate of 1.0 ml/ min. The detection wavelength was 284 nm with column temperature of 30°C. rrith phellodendrine chloride, gentiopicroside, paeoniflorin, tetrandrine and berberine hydrochloride as the object references, ten batches of Cangbaiqutong capsules were tested and analyzed by similarity comparison. Results fingerprint spectrum in Cangbaiqutong capsules had 20 common peaks in total, and characteristic spectra of Cortex Phellodendri, Gentiana, Radix Paeoniae Rubra and Radix Stephaniae Tetrandrae were found, while similarity of HPLC fingerprint was more than 0.9 among those batches of samples. Conclusion Using HPLC fingerprint can be used to evaluate the Cangbaiqutong capsules% quality in the round, which could improve the quality control standard in productive process of Cangbaiqutong capsules.

Simultaneous determination of seven constituents in Longsha Granules by dual-wavelength RP-HPLC / 中草药

Objective: To establish the RP-HPLC method for the simultaneous determination of phellodendrine, rutin, baicalin, baicalein, berberine hydrochloride, quercetin, and atractylodin in Longsha Granules. Methods: The seven components were measured on a C18 column (250 mm × 4.6 mm, 5 μm) of Kromasil with a gradient elution of acetonitrile -0.02% phosphoric acidsolution at the wavelengths of 205 nm (phellodendrine, rutin, baicalin, baicalein, and quercetin) and 340 nm (berberine hydrochloride and atractylodin), a flow rate of 1.0 mL/min, and the column temperature of 30 ℃. Results: Phellodendrine, rutin, baicalin, baicalein, berberine hydrochloride, quercetin, and atractylodin showed a good correlation in the ranges of 0.011-0.168 mg/mL (r = 0.9999), 0.010-0.160 mg/mL (r = 0.9999), 0.053-0.842 mg/mL (r = 0.9999), 0.014-0.228 mg/mL (r = 0.9998), 0.016-0.266 mg/mL (r = 0.9998), 0.009-0.144 mg/mL (r = 0.9999), and 0.002-0.024 mg/mL (r = 0.9997). The average recovery rates were 99.83%, 100.09%, 99.98%, 99.54%, 100.14%, 99.67%, and 100.06%. The RSD were 1.9%, 2.6%, 3.7%, 2.4%, 2.1%, 1.8%, and 3.2%. Conclusion: The method is simple, accurate, and excellent repetition, and can be used to control the quality of Longsha Granules.