Fingerprint establishment of Huangqin decoction and study on spectrum-effect relationship of its antidermatophytic activity in different phase states / 中国药房

OBJECTIVE To establish the fingerprint of Huangqin decoction （HQD）， to separate the phase states and screen the active phase states of antidermatophytic activity so as to study the spectrum-effect relationship. METHODS HPLC method was adopted using baicalin as reference， the fingerprints of 10 batches of HQD were drawn and the similarity evaluation was carried out using the Similarity Evaluation System of Chromatographic Fingerprint of TCM （2012 edition） to determine the common peak； the phase states of HQD were separated and characterized by high-speed centrifugation and membrane dialysis. The minimum inhibitory concentrations （MIC） of HQD and its different phase states against Trichophyton mentagrophytes were determined simultaneously. Using the peak area of 37 common peaks as independent variable， MIC as dependent variable， Pearson correlation analysis was performed by using SPSS 21.0 software. RESULTS A total of 37 common peaks were obtained in HPLC fingerprints of 10 batches of HQD， with the similarity higher than 0.99. Ten components were identified， such as albiflorin， paeoniflorin， liquiritin apioside， baicalin， melaleuca glycoside A， wogonoside， baicalein， glycyrrhizic acid， wogonin and oroxylin A. HQD was split into 3 phase states， such as precipitation phase （HQD-P）， solution phase （HQD-S） and nano phase （HQD-N）. The morphology of HQD-P was irregular granular， and the average particle size was 4.670-91.522 μm. The morphology of HQD-S was uniform flakes， and no particle size was detected. HQD-N was spherical in shape and the particle size was （129.0±12.9） nm. MIC values of each phase state of HQD against T. mentagrophytes in different phase states were HQD-N （4.64 mg/mL） ＜HQD （5.85 mg/mL） ＜HQD-P （7.37 mg/mL） ＜HQD-S （12.89 mg/mL） at the same dosage. Pearson correlation analysis showed that the peak area of 25 of the 37 common peaks （including identified components） was significantly negatively correlated with MIC （absolute values of correlation coefficient＞0.95 and P＜0.05）. CONCLUSIONS The chemical composition of 10 batches of HQD is consistent； HQD-N is the active phase state of HQD. Ten components such as paeoniflorin， liquiritin apioside and baicalin may be the main active components of HQD. The antidermatophytic effect of HQD is closely related to its component content and physical phase state.

Huangqin Decoction Attenuates DSS-Induced Mucosal Damage and Promotes Epithelial Repair via Inhibiting TNF-α-Induced NF-κB Activation / 中国结合医学杂志

OBJECTIVE@#To investigate the protective effect of Chinese herbal formula Huangqin Decoction (HQD) on ulcerative colitis mouse model induced by dextran sulphate sodium (DSS) and human intestinal epithelial cell injury induced by tumour necrosis factor-α (TNF-α).@*METHODS@#In vivo, 30 male C57BL/6 mice were divided into 5 groups using a random number table (n=6 per group), including control, DSS, 5-aminosalicylic acid (5-ASA), HQD low- (HQD-L) and high-dose (HQD-H) groups. The colitis mouse model was established by 3% (w/v) DSS water for 5 days. Meanwhile, mice in the HQD-L, HQD-H and 5-ASA groups were administrated with 100, 200 mg/kg HQD or 100 mg/kg 5-ASA, respectively, once daily by gavage. After 9 days of administration, the body weight, disease activity index (DAI) score and colon length of mice were measured, the pathological changes of colons were analyzed by hematoxylin-eosin staining (HE) staining, and the levels of serum interleukin (IL)-6, IL-1β and TNF-α were measured by enzyme linked immunosorbent assay. In vitro, the human colon epithelial normal cells (FHC cells) were exposed to HQD (0.6 mg/mL) for 12 h and then treated with TNF-α (10 ng/mL) for 24 h. The tight junction (TJ) protein expression levels of Claudin-4 and Occludin, and the protein phosphorylation levels of p65 and inhibitor of nuclear factor kappaB (NF-κB)-α (IκBα) were measured by Western blot.@*RESULTS@#In vivo, compared with the DSS group, HQD-H treatment attenuated the weight loss and reduced DAI score of mice on the 8th day (P<0.05). Moreover, HQD-H treatment ameliorated the colon shortening in the DSS-induced colitis mice (P<0.05). HE staining showed HQD attenuated the pathological changes of colitis mice, and the histological scores of HQD-H and 5-ASA groups were significantly decreased compared with the DSS group (P<0.05). Meanwhile, HQD-H and 5-ASA significantly decreased the serum IL-1β, IL-6 and TNF-α levels of mice (P<0.05). In vitro experiments showed that HQD up-regulated Occludin and Claudin-4 protein expressions and inhibited p-p65 and p-IκBα levels in FHC cells compared with the TNF-α group (P<0.05).@*CONCLUSION@#HQD significantly relieved the symptoms in DSS-induced colitis mice by inhibiting pro-inflammatory cytokines expression and maintained the homeostasis of TJ protein in FHC cells by suppressing TNF-α-induced NF-κB activation.

Pharmacological effects of Huangqin Decoction prepared from roots of multi-originated peony on ulcerative colitis in mice:a comparative study / 中国中药杂志

Huangqin Decoction(HQD) is a classic prescription for treating dysentery in the Treatise on Cold Damage and now is mainly used for the treatment of ulcerative colitis(UC). Since there are no requirements on specific Paeonia species, both Paeoniae Radix Alba(white peony root, WPR) and Paeoniae Radix Rubra(red peony root, RPR) are clinically used in HQD now. Although the two types of peony roots are close in origin and similar in primary components, the medicinal properties and efficacies are different. Furthermore, the systematic comparative analysis on the efficacy differences in treating UC of HQD with the roots of multi-originated peony has been seldom reported. This study compared and evaluated the pharmacological effects of HQD prepared from the roots of multi-originated peony, including WPR, RPR-l(derived from P. lactiflora), and RPR-v(derived from P. veitchii) based on the mouse model of UC induced by dextran sodium sulfate(DSS) by animal behaviors, pathological section(colon), and cytokine expression(IL-1β and IL-6), aiming to provide evidence for the identification of the original resource of peony root in HQD. The results indicated that all HQD samples prepared from WPR, RPR-l, and RPR-v could improve the symptoms of UC. Compared with the HQD-WPR, HQD-RPR-l and HQD-RPR-v were significantly different in weight loss, colon length, and disease activity index(DAI) score, but there was no significant difference between HQD-RPR-l and HQD-RPR-v. Moreover, HQD-RPR-v exhibited the most significant improvement in the pathological morphology of colonic tissue and mucosal defects. According to the previous comparative analysis of chemical profiling and content distribution of HQD prepared from the roots of multi-originated peony, RPR-v in HQD was potent in protecting against UC, which was presumedly attributed to a large number of monoterpene glycosides and galloyl glucoses. This study provided a scientific basis for the determination of peony root in HQD and its clinical medication.

Effect of Huangqin Decoction on pyroptosis pathway of NLRP3/caspase-1 in mice with ulcerative colitis / 中国中药杂志

To explore the effect of Huangqin Decoction on ulcerative colitis(UC) pyroptosis, and to explain the mechanism of pyroptosis based on NOD-like receptor thermoprotein domain 3(NLRP3)/cysteine proteinase 1(caspase-1) pathway. The animal model of UC induced with 3% dextran sodium sulfate(DSS) was established. The experimental animals were divided into control group, model group, low-dose(4.55 g·kg~(-1)), medium-dose(9.1 g·kg~(-1)) and high-dose(18.2 g·kg~(-1)) Huangqin Decoction groups and salazosulfapyridine group(0.45 g·kg~(-1)). While modeling, intragastric administration was given for 7 consecutive days. On the 8 th day, the mice were euthanized, the colon length was collected, and the histopathological changes were observed by HE staining. The content of interleukin-18(IL-18) was observed by ELISA. The content of lactatedehydrogenase(LDH)was determined by microplate method. TUNEL assay kit was used to detect the cell death. The immunohistochemical staining was used to detect the expressions of NLRP3 and apoptosis-associated speck-like protein containing a CARD(ASC). Western blot was used to detect the expressions of interleukin-1β(IL-1β), caspase-1 and gasdermin D(GSDMD).The experimental study showed that compared with normal group, the LDH content, TUNEL positive staining, inflammatory factors(IL-18, IL-1β), and proteins associated with pyroptosis were significantly increased(P<0.05). Compared with model control group, the LDH content, TUNEL positive staining, inflammatory factors(IL-18, IL-1β), and proteins associated with pyroptosis were decreased, and these results were more significant in high-dose groups(P<0.05). The results of HE staining showed that Huangqin Decoction could improve the pathological changes of colon. Huangqin Decoction could inhibit UC cell pyroptosis, and the mechanism may be closely related to NLRP3/caspase-1 signaling pathway.

Potential mechanism of Huangqin decoction for the treatment of ulcerative colitis based on network pharmacology / 药学学报

To study the mechanism of Huangqin decoction (HQT) in the treatment of ulcerative colitis (UC) by using network pharmacology, chemical components and targets related to the four herbs of Chinese meteria medical in HQT were searched through the traditional Chinese medicine systems pharmacology database and analysis platform (TCMSP) to construct the interaction network diagram of the target point of the compounds. The UC-related targets were screened through OMIM, TTD, and GeneCard databases. The compound-target network was constructed using Cytoseape_v3.7.1 software; based on the STRING database, a target interaction network for HQT for UC was constructed, and the core target of HQT for UC was selected based on topological parameters. GO (gene ontology) biological process enrichment analysis and KEGG (KEGG pathway analysis) pathway annotation analysis were performed on the disease and drug intersection targets using the R package clusterprofile version 3.12.0 in Bioconductor. The HQT compound-UC target network contains 128 compounds and corresponding targets 141. The core targets are AKTI, IL6, PTGS2, IL10, IL1β and so on. GO functional enrichment analysis yielded 151 GO terms, and KEGG pathway enrichment screening resulted in 33 associations with UC, mainly involving PI3K-AKT signaling pathway, NF-kappa B signaling pathway, TNF signaling pathway, Toll-like receptor signaling pathway and so on. The synergetic effect of HQT with multi-components and multi-pathway was confirmed by network pharmacology, and the main possible mechanism of HQT in treating UC was predicted, which lay a foundation for the identification of effective components, the mechanism of action, and clinical application.

Interactions of component (group) compatibility of Sanwu Huangqin Decoction in incubation model of rat liver microsomes / 中成药

AIM To study the interactions of component (group) compatibility of Sanwu Huangqin Decoction in incubation model of rat liver microsomes.METHODS With maackiain and ephedrine as internal standards,HPLC was adopted in the simultaneous content determination of total tlavonoids (baicalin,wogonoside,baicalein,wogonin and oroxylin A) from Scutellariae Radix and total alkaloids (oxymatrine,oxysophocarpine and matrine)from Sophorae flavescentis Radix in whole prescription component (group) compatibility and single medicinal material part (group) at seven time points (0,15,30,45,60,90 and 120 min),followed by the calculation of their in vitro metabolic rates.RESULTS In whole prescription component (group) compatibility,the contents of baicalin and wogonoside were first increased (0-0.5 h) and then decreased (0.5-2.0 h),those of baicalein,wogonin and oroxylin A showed increasing trends (more obvious within 0.5 h).The stable and gende metabolisms of various alkaloids reached balance within 20 min.CONCLUSION The bioavailability improvement and efficacy enhancement of total flavonoids from Scutellariae Radix may attribute to the compatibility with total alkaloids from Sophorae flavescentis Radix and total polysaccharides from Rehmanniae Radix in Sanwu Huangqin Decoction

Interactions of component (group) compatibility of Sanwu Huangqin Decoction in incubation model of rat liver microsomes / 中成药

AIM To study the interactions of component (group) compatibility of Sanwu Huangqin Decoction in incubation model of rat liver microsomes.METHODS With maackiain and ephedrine as internal standards,HPLC was adopted in the simultaneous content determination of total tlavonoids (baicalin,wogonoside,baicalein,wogonin and oroxylin A) from Scutellariae Radix and total alkaloids (oxymatrine,oxysophocarpine and matrine)from Sophorae flavescentis Radix in whole prescription component (group) compatibility and single medicinal material part (group) at seven time points (0,15,30,45,60,90 and 120 min),followed by the calculation of their in vitro metabolic rates.RESULTS In whole prescription component (group) compatibility,the contents of baicalin and wogonoside were first increased (0-0.5 h) and then decreased (0.5-2.0 h),those of baicalein,wogonin and oroxylin A showed increasing trends (more obvious within 0.5 h).The stable and gende metabolisms of various alkaloids reached balance within 20 min.CONCLUSION The bioavailability improvement and efficacy enhancement of total flavonoids from Scutellariae Radix may attribute to the compatibility with total alkaloids from Sophorae flavescentis Radix and total polysaccharides from Rehmanniae Radix in Sanwu Huangqin Decoction

Progress in Pharmacodynamic Material Basis and Treatment for Gastrointestinal Diseases of Huangqin Decoction / 现代生物医学进展

Huangqin Decoction is a classic recipe for treatment of diarrhea from Treatise on Exogenous Febrile Disease,and consists of Scutellaria baicalensis,Paeonia lactiflora,Glycyrrhiza uralensis and Ziziphus jujuba.Huangqin Decoction has been used for nearly 1800 years for the treatment of gastrointestinal ailments,including dysentery,diarrhea and so on.Treatment of gastrointestinal disease by traditional Chinese medicine is considered to be safer than western medicine.This paper presented a review ofpharmacodynamic material basis of Huangqin Decoction,its advances in treating ulcerative colitis and reduction of the gastrointestinal side effects arising from chemotherapy drugs according to related literature.It also reviewed the deficiencies in present research,outlined the future direction and provided reference for further study.

Comparison of baicalin,paeoniflorin and glycyrrhetic acid in different decoctions of Huangqin Decoction / 中成药

AIM: To compare the contents of baicalin,paeoniflorin and glycyrrhetic acid in the seperated and mixed decoction of Huangqin Decoction(Radix Scutellariae,Radix Paeoniae alba,Radix et Rhizoma Glycyrrhizae and Fructus Jujubae). METHODS: The contents of baicalin,paeoniflorin and glycyrrhetic acid were analyzed by HPLC.Chromatographic conditions included: Hypersil BDS C_(18) column and the mobile phase consisted of a mixture of methanol-water-phosphoric acid(47(∶)53(∶)0.2),acetonitrile-water-phosphoric acid(18(∶)82(∶)0.1) and methanol-water-phosphate buffer solution(70(∶)29(∶)1).Baicalin,paeoniflorin and glycyrrhetic acid were detected at 280 nm,230 nm and 250 nm respectively. RESULTS: The linear range of baicalin,paeoniflorin and glycyrrhetic acid were 2.88—72.00 ?g/mL,2.85—22.80 ?g/mL and 4.16—52.00 ?g/mL respectively.The contents of baicalin in the mixed decoction was higher than that in the seperated decoction.The contents of paeoniflorin and glycyrrhetic acid in the mixed decoction were almost as same as that in seperated decoction. CONCLUSION: The contents of active ingredients in the mixed and seperated decoction are different.