Wogonin Research Articles

Wogonin Alleviates Streptozotocin-stimulated Diabetic Nephropathy Through Its Antidiabetic, Antidyslipidemic, and Antioxidative Effects in Rats

Background Diabetes mellitus (DM) is a prolonged endocrine syndrome recognized by defective metabolism of biomolecules on account of flaws in insulin resistance. Diabetic nephropathy (DN) is a central microvascular complication of DM with inadequate treatment alternatives. Wogonin (WOG) is a pharmacologically active flavonoid isolated from the roots of Scutellaria baicalensis Georgi, which exerts strong renoprotection. However, the defensive mechanism of WOG against DN is not completely illuminated. Purpose This study investigates the role of WOG in streptozotocin (STZ)-induced DN in high-sucrose-high-fat fed rats. Methods DM was induced in experimental rats by a solo dosage of STZ (40 mg/kg) administered intraperitoneally. All rats were allocated into five sets, namely, normal, diabetic control (DC), metformin (150 mg/kg), and WOG (25 and 50 mg/kg) treated orally for 2 months. Results The antidiabetic and renoprotective effects of WOG were assessed by analyzing lipid profiles, glucose, inflammatory mediators, oxidative stress markers, renal functional parameters, gain in body weight, and histopathology of the kidney. In the current study, WOG could improve DM-induced metabolic alterations, dyslipidemia, kidney dysfunction, and inflammatory and oxidative prominence over its antidiabetic, antioxidative, antihyperlipidemic, and anti-inflammatory actions. Conclusion Thus, WOG is employed as a nephroprotective agent in DC rats.

Wogonoside alleviates microglia-mediated neuroinflammation via TLR4/MyD88/NF-κB signaling axis after spinal cord injury

Wogonoside (WG) is a natural flavonoid extracted from Scutellariae Radix, recognized for its established anti-inflammatory properties. However, the role of WG in the context of neuroinflammation after spinal cord injury (SCI) remains inadequately elucidated. This study employed in silico, in vitro, and in vivo methodologies to investigate the impact of WG on microglia-mediated neuroinflammation after SCI. In the in silico experiment, we identified 15 potential target genes of WG associated with SCI. These genes were linked to the regulation of inflammatory response and immune defense. Molecular docking maps revealed toll-like receptor 4 as a molecular target for WG, demonstrating binding through a hydrogen bond (Lys263, Ser120). In lipopolysaccharide-stimulated BV2 cells and SCI mice, WG significantly attenuated microglial activation and facilitated a phenotype shift from M1 to M2. This was evidenced by the reversal of the increased expressions of Iba1, GFAP, and iNOS, as well as the decreased expression of Arg1. WG also suppressed the production of pro-inflammatory mediators (NO, TNF-α, IL-6, IL-1α, IL-1β, C1q). WG exerted these effects by suppressing the TLR4/MyD88/NF-κB signaling axis in microglia. Furthermore, by reducing levels of TNF-α, IL-1α, and C1q in supernatant of LPS-induced microglia, WG indirectly induced astrocytes change to A2 phenotype, evidenced by transcriptome sequencing result of primary mouse astrocytes. All these events above collectively created a favorable microenvironment, contributing to a significant alleviation of weight loss and neuronal damage at the lesion site of SCI mice. Our findings substantiate the efficacy of WG in mitigating neuroinflammation after SCI, thereby warranting further exploration.

Identification of novel targets and mechanisms of wogonin on lung cancer, bladder cancer, and colon cancer

Wogonin (WOG) has been demonstrated to have anti-cancer activity, but the mechanisms remain unclear. In this study, new targets of WOG were predicted for lung cancer, bladder cancer, and colon cancer by using bioinformatics methods. WOG might primarily suppress cancers via regulating arachidonic acid, Ras, MAPK, linoleic acid, PI3K-Akt, and folate biosynthesis pathways. 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay showed that WOG inhibited the proliferation of A549 cells. Real-time quantitative reverse transcription PCR (RT-qPCR) results indicated that anti-lung cancer effect of WOG was achieved by regulating the expression of 18 target genes, including AKR1B10, AKR1C3, BDNF, CAV1, CXCL2, CYP2B6, CYP4F3, DAO, EGF, ENO3, IL6, PLA2G1B, PLA2G2F, PLA2G4A, PTGES, SLCO1B1, SLCO1B3, and TFAP2A. The Kaplan-Meier survival curves further confirmed that DAO, PLA2G1B, SLCO1B3 and TFAP2A were essential targets via which WOG affected lung cancer survival. Moreover, BDNF, FGF2, and PTGS1 were predicted to be the targets via which WOG alleviated cancer proliferation and invasion in bladder cancer. As for colon cancer, WOG might induce autophagy and inhibit proliferation by down-regulating NTF4 and TH. The study will provide clue for using WOG as a promising antineoplastic agent in basic and translational research, and bring light to the application of herbs containing WOG as food supplements.

Wogonin Diminishes Radioresistance of Breast Cancer via Inhibition of the Nrf2/HIF-1[Formula: see text] Pathway.

Radiotherapy plays a crucial role in the multimodal treatment of breast cancer. However, radioresistance poses a significant challenge to its effectiveness, hindering successful cancer therapy. Emerging evidence indicates that Nrf2 and HIF-1[Formula: see text] are critical regulators of cellular anti-oxidant responses and that their overexpression significantly promotes radioresistance. Wogonin (WG), the primary component isolated from Scutellaria baicalensis, exhibits potential antitumor and reversal of multidrug resistance activities. Nevertheless, the role of WG in radioresistance remains unclear. This study aims to explore the effects of WG on the radioresistance of breast cancer. Our results indicate that Nrf2 and HIF-1[Formula: see text] overexpression was observed in breast cancer tissues and was correlated with the histological grading of the disease. Radiation further increased the levels of Nrf2 and HIF-1[Formula: see text] in breast cancer cells. However, WG demonstrated the ability to induce cell apoptosis and reverse radioresistance by inhibiting the Nrf2/HIF-1[Formula: see text] pathway. These effects were also confirmed in xenograft mice models. Mechanistically, WG enhanced the level of the Nrf2 inhibitor Keap1 through reducing CpG methylation in the promoter region of the Keap1 gene. Consequently, the Nrf2/HIF-1[Formula: see text] pathway, along with the Nrf2- and HIF-1[Formula: see text]-dependent protective responses, were suppressed. Taken together, our findings demonstrate that WG can epigenetically regulate the Keap1 gene, inhibit the Nrf2/HIF-1[Formula: see text] pathway, induce apoptosis in breast cancer cells, and diminish acquired radioresistance. This study offers potential strategies to overcome the limitations of current radiotherapy for breast cancer.

Discovery of conversion driven by β-glucuronidase from flavone glycoside to aglycone and application in identifying the raw Scutellariae Radix

Scutellariae Radix (SR), the dried root of Scutellaria baicalensis Georgia, is a famous Chinese materia medica that has been widely employed. Raw Scutellariae Radix (RSR), steamed Scutellariae Radix (SSR), and wine Scutellariae Radix (WSR) are adopted for use in clinical practice. Because of their easily confused appearance, they are always misused. Aiming at this problem, an ultra performance liquid chromatography coupled with photodiode array detector (UPLC-PDA) method was established to survey misuse of the RSR and the processed SR (SSR and WSR) in the market by employing baicalin (BC), wogonoside (WS), baicalein (BN), and wogonin (WN) as quality indicators. Fortunately, β -glucuronidase, which mediates conversion from flavone glycoside to aglycone, was identified in the RSR samples by the sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. The significant production of BN and WN was witnessed in the RSR samples, which did not occur in the SSR and WSR samples in virtue of the inactivated β -glucuronidase. Besides, the different capacities of β -glucuronidase were evaluated in the tested samples. In general, we provided the first evidence to scientifically identify RSR from SSR and WSR.

Wogonoside attenuates liver fibrosis by triggering hepatic stellate cell ferroptosis through SOCS1/P53/SLC7A11 pathway.

Wogonoside (WG) is a flavonoid chemical component extracted from Scutellaria baicalensis, which exerts therapeutic effects on liver diseases. Ferroptosis, a novel form of programmed cell death, regulates diverse physiological/pathological processes. In this study, we attempted to investigate a novel mechanism by which WG mitigates liver fibrosis by inducing ferroptosis in hepatic stellate cells (HSCs). A CCl4 -induced mouse liver fibrosis model and a rat HSC line were employed for in vivo and in vitro experiments, both treated with WG. Firstly, the levels of the fibrotic markers α-smooth muscle actin (α-SMA) and α1(I)collagen (COL1α1) were effectively decreased by WG in CCl4 -induced mice and HSC-T6 cells. Additionally, mitochondrial condensation and mitochondrial ridge breakage were observed in WG-treated HSC-T6 cells. Furthermore, ferroptotic events including depletion of SLC7A11, GPX4 and GSH, and accumulation of iron, ROS and MDA were discovered in WG-treated HSC-T6 cells. Intriguingly, these ferroptotic events did not appear in hepatocytes or macrophages. WG-elicited HSC ferroptosis and ECM reduction were dramatically abrogated by ferrostatin-1 (Fer-1), a ferroptosis inhibitor. Importantly, our results confirm that SOCS1/P53/SLC7A11 is a signaling pathway which promotes WG attenuation of liver fibrosis. On the contrary, WG mitigated liver fibrosis and inducted HSC-T6 cell ferroptosis were hindered by SOCS1 siRNA and pifithrin-α (PFT-α). These findings demonstrate that SOCS1/P53/SLC7A11-mediated HSC ferroptosis is associated with WG alleviating liver fibrosis, which provides a new clue for the treatment of liver fibrosis.

Inhibition of Radix Scutellariae flavones on carboxylesterase mediated activations of prodrugs

AimsCarboxylesterase (CES) plays an essential role in the hydrolysis of ester prodrugs. Our study explored the inhibitions of Radix Scutellariae flavones, including baicalein (B), baicalin (BG), wogonin (W), wogonoside (WG), oroxylin A (OXA) and oroxylin A-7-O-glucuronide (OAG), on CES-mediated hydrolysis of seven prodrugs (capecitabine, clopidogrel, mycophenolate mofetil, dabigatran etexilate, acetylsalicylic acid, prasugrel and irinotecan). Main methodsIn vitro screenings were developed by incubating the flavones with prodrugs in rat plasma, intestine S9 and liver S9. Docking simulations were conducted using AMDock v1.5.2. In vivo evaluations were performed in rats co-administered with the selected flavone and prodrug via oral gavage/intravenous administration for five consecutive days. Key findingsThe in vitro investigation showed that B and OXA demonstrated strongest inhibitions on the hydrolysis of irinotecan followed by dabigatran in rat plasma, intestine S9 and liver S9. Consistent results showed in the molecular docking analyses. Additionally, in rats receiving irinotecan, B/OXA intravenous and oral pre-treatments both led to reduction trends on the active metabolite SN-38 formation in plasma. Besides, significant decreases of SN-38/irinotecan plasma concentration ratios were found in the B/OXA oral pre-treatment group with quicker and stronger inhibition potential in OXA pre-treatment than that from B pre-treatment. OXA oral pre-treatment was also found to be able to significantly inhibit intestinal CES2 activities at 0.5 h and 5 h after irinotecan administration. SignificanceOur current findings for the first time alert on potential CES-mediated HDIs between RS flavones and prodrugs, which provide a constructive information referring to rational drug combinations in clinical practice.

Wogonin and its analogs for the prevention and treatment of cancer: A systematic review.

The medicinal plant Scutellaria baicalensis, commonly known as Chinese skullcap or Huang-Qin, has been used as a traditional medicine for several thousand years. The roots of this plant contain bioactive compounds, such as wogonin (WOG), wogonoside, baicalein, and baicalin. The aim of this article is to evaluate the therapeutic potential and mechanisms of action of WOG against different cancers. Numerous in vitro and in vivo studies have revealed that WOG exerts immense therapeutic potential against bladder cancer, breast cancer, cholangiocarcinoma, cervical cancer, colorectal cancer, gallbladder cancer, gastric cancer, glioblastoma, head and neck cancer, hepatic cancer, leukemia, lung cancer, lymphoma, melanoma, multiple myeloma, neuroblastoma, osteosarcoma, ovarian cancer, pancreatic cancer, prostate cancer, and renal cancer by regulating various cell signaling pathways. WOG, in combination with established chemotherapeutic drugs, improves the efficacy of treatment and lowers toxicity. Nevertheless, human trials are warranted to validate these findings. Numerous preclinical studies, combined with an extensive margin of safety and no severe side effects, underscore WOG's therapeutic potential as an anticancer drug. These studies propound the use of WOG as a potential anticancer candidate; however, further high-quality studies are required to firmly establish the clinical efficacy of WOG for the prevention and treatment of human malignancies.

Wogonin reduces cardiomyocyte apoptosis from mitochondrial release of cytochrome c to improve doxorubicin-induced cardiotoxicity.

Doxorubicin (DOX) has powerful anticancer properties, but its clinical application is affected by its serious cardiotoxicity. Wogonin (WG) has been shown to have marked cardiovascular protection potential. However, it is not known whether this potential can protect the heart from DOX damage. The aim of the present study was to investigate whether WG could ameliorate the cardiotoxicity of DOX. DOX and WG were used to establish a model of cardiac damage. Echocardiography, brain natriuretic peptide, creatine kinase MB and cardiac troponin T were used to detect the degree of cardiac damage. The levels of superoxide dismutase, malondialdehyde, glutathione and catalase in serum were measured to observed oxidative stress state. The mRNA levels of cyclophilin D, voltage-dependent anion-selective channel 1 and adenine nucleotide transporter 1 were detected by reverse transcription-quantitative PCR. Western blotting was used to detect the expression of cytochrome c in mitochondria and cytoplasm and cleaved-caspase-9 and pro/cleaved-caspase-3 in cytoplasm in cardiac tissue and primary cardiomyocytes to verify the related signaling pathways. DOX rats showed a series of cardiac damage. However, these damages were alleviated following WG treatment. Further studies showed that WG antagonized DOX cardiotoxicity through inhibiting the release of cytochrome c. WG protected rat heart from DOX damage. The mechanism may be closely related to inhibiting the release of cytochrome c from mitochondria and reducing cardiomyocyte apoptosis caused by caspase activation.

Exploring the synergistic mechanism of Gegen Qinlian Decoction on the Wnt signaling pathway using an integrated strategy of network pharmacology and RNA-seq

Ethnopharmacological relevanceGegen Qinlian Decoction (GQD) (including: Puerariae lobatae (Willd.) Ohwi, radix; (short for Gengen) Glycyrrhiza uralensis Fisch., root and rhizome (short for Gancao), honeyed; Coptis chinensis Franch., rhizome (short for Huanglian); Scutellaria baicalensis Georgi, radix, boiled (short for S. baicalensis) has been widely used to treat inflammatory bowel disease (IBD) and colorectal cancer (CRC). To explore compatibility mechanism of GQD could be of advantage to investigate the complex principle of TCM, which might be conducive to the exploration of the modernization of TCM. Aim of reviewIn this study, a strategy based on system pharmacology was constructed to uncover the multi-target regulation and compatibility mechanism of GQD on the Wnt signaling pathways. Material and methodsThe pharmacological network of GQD was constructed by TCMSP, DAVID, Uniprote database. The cell growth inhibitory effects of puerarin (PUE), wogonin (WOG), berberine (BER), and glycyrrhetinic acid (GLY) on SW480 cells were assessed using CCK-8 assay. The multi-target regulation and compatibility mechanism of combination PUE with GLY were examined by RNA-seq, HPLC-QQQ/MS, qRT- PCR and Western blot analysis. ResultsNetwork pharmacology analysis indicated that PUE, WOG, BER and GLY were the active components in GQD and had a synergistic effect on the targets of the Wnt signaling pathway. Additionally, pharmacological experiments revealed that WOG, BER, and GLY inhibited activity of colorectal cancer (CRC) cell lines SW480 cells, and that PUE only exhibited effective antitumour activity when combined with GLY. CTNNB1, CCND1 and SMAD4 were identified as synergistic targets inhibited by PUE-GLY. Moreover, PUE-GLY could influence the Wnt signaling pathway by upregulating GSK3B and downregulating CTNNB1 synergistically. It also showed that GLY could effectively increase the intracellular content of PUE based on HPLC-QQQ/MS analysis, and this process was achieved by influencing the targets of the membrane's pathway, such as cell adhesion molecules, focal adhesion, and tight junctions. ConclusionGLY was revealed a multi-target mechanism, which could downregulate CTNNB1 as the active component and intervene in membrane proteins (CDH1, CADM1, ITGB2, ICAM1, ITGA1) as ‘guide’ in the formulae. Moreover, the mechanism of synergistic antitumour action of PUE (the active component of Monarch drug) and GLY (the active component of Guide drug) on the Wnt signaling pathway was explored systematically. It was a promising breakthrough for elucidating the scientific connotation of the compatibility of TCM formulae and provide a valuable and practicable methodology for clarifying the mechanisms of TCM.

Wogonoside attenuates the articular cartilage injury and the infiltration of Th1/Th2-type cytokines in papain-induced osteoarthritis in rat model via inhibiting the NF-κB and ERK1/2 activation

Objects Osteoarthritis is the most common joint disease and a major cause of functional limitation and pain in adults. This study aims to investigate the effect of wogonoside (WOG) on the progression of knee osteoarthritis (KOA) in model rats. Materials and methods Rats KOA models were established and treated with different doses of WOG (10 mg/kg, 20 mg/kg and 30 mg/kg). The degree of cartilage injury was detected by Mankin scores via HE/Alcian blue staining. The levels of IFN-γ and IL-4 in peripheral blood and synovial fluid and the Th1/Th2 ratio were detected by flow cytometry. The model mice were injected with NF-κB p65 or ERK1/2 inhibitors or activators to further investigate the effect of WOG on KOA. Results WOG significantly improved cartilage tissue damage and reduced the Mankins score. WOG down-regulated the level of IFN-γ while up-regulated the expression of IL-4, which maintained the balance of Th1/Th2 cells. Further studies showed that the expression of NF-κB p65, phosphorylated p65, cytoplasmic ERK1/2 and nuclear ERK1/2 were all inhibited by WOG. The results of reverse verification experiments showed that the activator of NF-κB p65 and ERK1/2 weakened the protective effect of WOG on KOA, and the inhibitor of NF-κB p65ERK1/2 enhanced the protective effect of WOG on KOA. Conclusions WOG inhibited the activation of NF-κB and ERK1/2 to alleviate the articular cartilage injury and Th1/th2 cytokine infiltration in KOA rats.

Inhibitory effects of Coptidis Rhizoma on the intestinal absorption and metabolism of Scutellariae Radix

Ethnopharmacological relevanceThe Scutellariae Radix (SR) and Coptidis Rhizoma (CR) herb couple is widely used in traditional Chinese medicine prescriptions for the treatment of diabetes mellitus due to its interaction and synergistic effect compared to either herb alone, but the underlying mechanism of interaction between these herbs is unclear. This study aimed to investigate the effects of CR on the metabolism and absorption of SR. Materials and methodsAfter rats were treated with normal saline (NS group) or the CR extract (CR-treated group) for seven consecutive days, the intestinal flora was extracted from rat faeces for a co-incubation with the SR extract to investigate the metabolism of SR flavonoids, and a non-everted gut sac was prepared in vitro to evaluate the intestinal absorption of the SR extract. The components of the SR extract, the metabolites of the SR extract that was co-incubated with intestinal flora, and the dialysate acquired from non-everted gut sacs were identified and determined by an HPLC-MS/MS method. The absorption rate constant (Ka) and the apparent permeability (Papp) of each compound were calculated, and the effects of CR on the metabolism and absorption of flavonoids in SR were evaluated, by comparison the Ka and Papp between two groups using Student's t-test. ResultsTwenty-nine flavonoids were detected and identified in the SR extract, including 16 glycosides and 13 aglycones. In the co-incubation with the intestinal flora, differences in metabolite classes were not observed between the NS group and CR-treated group; however, the metabolic rates of 17 flavonoids in the CR-treated group were significantly higher than the NS group. The Papp of 11 compounds (4 glycosides and 7 aglycones) across the gut sac were greater than 2 × 10−5 cm/s in both groups, while the Papp values of 7 compounds including wogonoside (WG) and other aglycones were significantly decreased in the CR-treated group. ConclusionBased on these results, CR decreased the metabolism and absorption of SR flavonoids, and exerted much greater inhibitory effects on aglycones than glycosides, which may be one of the potential mechanisms underlying the therapeutic effects of the combination of SR and CR on diabetes mellitus.

Disruption of SSBs repair to combat platinum resistance via the JWA-targeted Pt(IV) prodrug conjugated with a wogonin derivative.

An octahedral Pt (IV) prodrug, Cis-wog, containing a wogonin derivative as a bioactive axial ligand was designed and prepared to suppress DDR (DNA damage repair)-related proteins. In vitro biological studies indicated that a Pt (IV) prodrug with axially functional groups (Cis-wog) showed cytotoxicity superior to cisplatin and reversed its resistance against two pairs of cisplatin sensitive and resistant cell lines. Further mechanistic research revealed that the powerful antitumor activity of Cis-wog resulted from its suppression of JWA and its multi-interaction with XRCC1 to repair DNA single strand breaks (SSBs) caused by the introduction of wogonin. It is concluded that Cis-wog is a promising cytotoxic agent, which could be used for enhancing the antitumor activity of its corresponding Pt(II)-based drugs and reversing cisplatin resistance via decaying JWA-mediated SSBs repair pathways and inducing apoptosis.

The parametric measurementof physico-chemical parameter and three types of effective components in Jiedu-Zhixue Decoction

Objective To set up a method to measure the content of baicaline, wogonoside, and paeonol in Jiedu-Zhixue Decoction extract and to measure their physico-chemical parameters. Methods By using HPLC analytical methods to measure the content of baicaline, wogonoside, and paeonol in Jiedu-Zhixue Decoction extract. The RP18 column was adopted, gradient eluted with mobile phase of acetonitrile-1% phosphoric acid water, flow rate was 1 ml/min, detection wavelength was 274 nm, and the column temperature was 30 ℃. The method quantitatively analyze the compound extracts of baicaline, wogonoside and paeonol were used. Results The method isrepeatable, stable with good recorey rates. The calibration curves of baicaline was in good linearity over the range of 0.191 2-1.147 2 μg. The calibration curves of wogonoside was in good linearity over the range of 0.041 5-0.207 4 μg. The calibration curves of paeonol was in good linearity over the range of 0.019 5-0.156 3 μg. There was no significant difference among the component of 3 different batches of Jiedu-Zhixue Decoction extract, and the pH value range was 5.240-5.753, relative density range was 0.846-0.889. Conclusions The method used in this stydy to measure the effective components of Jiedu-Zhixue Decoction extract was stable and feasible, which is suitable for quality control of Jiedu-Zhixue Decoction. Key words: Dosage forms improving; Jiedu-Zhixue Decoction; Baicaline; Wogonoside; Paeonol; Physico-chemical parameter; High performance liquid chromatography; Quality control

Quantitative Analysis of Twelve Active Components Combined With Chromatographic Fingerprint for Comprehensive Evaluation of Qinma Prescription by Ultra-Performance Liquid Chromatography Coupled With Diode Array Detection.

A combination method of ultra-performance liquid chromatography (UPLC) coupled with diode array detection has been developed for quality evaluation of Qinma prescription (QMP), based on chromatographic fingerprint technology with the similarity analysis (SA) and the quantitative analysis of 12 components by hierarchical cluster analysis (HCA). The established method has been validated by linearity, precision, repeatability, stability and recovery tests. The UPLC fingerprints with 17 common peaks of 5 QMP samples prepared by different extraction methods including water decoction extraction, water extraction-ethanol precipitation method, ethanol reflux extraction, ethanol extraction-water precipitation method and methanol ultrasonic extraction were obtained, and the SA results indicated that similarity index was greatly influenced by the large peak. The similarity index ranged from 0.816 to 0.999 basing on 17 peaks, which has been decreased to 0.683-0.999 basing on 16 peaks without the large peak of baicalin (BA). The results of simultaneous quantification of 12 components in these 5 QMP samples proved that BA, gallic acid (GA), wogonoside (WOG) and gentiopicroside (GEN) were the major ingredients in QMP with high contents >1.44 (mg/g), indicating that ethanol reflux was the most effective extraction method. Integrating fingerprint analysis, simultaneous determination and HCA, the established method is rapid, sensitive, accurate and readily applicable. All the results indicated that the combination method can control the quality of QMP and its related traditional Chinese medicinal compounds more comprehensively and scientifically.

Topical Application of Wogonin Provides a Novel Treatment of Knee Osteoarthritis

Osteoarthritis (OA) is a degenerative joint disease characterized by joint pain, decreased functional mobility, and deformation of articular cartilage and consequentially diminishing quality of life. Wogonin (WG), a compound extracted from the Skullcap Baicalensis plant, has been shown to have anti‐inflammatory effects on the Toll‐like Receptor 4 (TLR‐4) inflammatory pathway and antioxidant properties via Nrf2. We have shown that TLR‐4 is a major receptor for pain in OA. In our study, we examined the pain reducing, anti‐inflammatory, and chondroprotective effects of WG when applied as a topical cream (TC). After destabilizing the menisci of mice to induce OA, we examined the severity and progression of OA with and without the topical application of WG. Using a running wheel to track mice activity, we found that mice with WG treatment were statistically more active than mice without WG treatment. OA progression analyzed using Modified Mankin and OARSI scoring showed a significant attenuation of OA severity among mice treated with WG. Immunohistochemistry revealed a significant decrease in protein expression of TGF‐β1, but no significant differences were found among the biomarkers HTRA1, MMP‐13, or BMP‐2, suggesting an alternative pathway of cartilage rescue.Support or Funding InformationBYU Office of Research and Creative WorksThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Simultaneous determination of five compounds in Ganmaoning granules by UPLC-MS/MS

Objective To establish a method for the simultaneous determination of adenosine, chlorogenic acid, caffeic acid, baicalin and wogonin in Ganmaoning granules. Methods The UPLC-MS/MS method was adopted. The separation was performed on ZOBAX SB C18 (2.1 mm×150 mm, 5 μm) column with mobile phase consisted of methanol-2 mmol/L ammonium acetate aqueous solution (isocratic elution) at the flow rate of 0.2 ml/min. The column temperature was set at 35 ℃, and the injection volume was 2 μl. The electrospray ionization source (ESI) was used. Multiple reaction monitoring (MRM) mode was adopted, using positive and negative ions scanning. The ion source temperature was 300 ℃, the drying gas temperature was 400 ℃, the drying gas flow was 20 L/min, the atomizing gas pressure was 55 psi, and the capillary voltage was 4000 V. Results The linear ranges of adenosine, chlorogenic acid, caffeic acid, baicalin and wogonin in Ganmaoling granules were 0.20-12.80 ng (r=0.999 6), 1.00-64.00 ng (r=0.999 8), 0.40-25.60 ng (r=0.999 6), 4.00-256.00 ng (r=0.999 2), 0.20-12.8 ng (r=0.999 1), which showed the linear relationship was good. The limits of detection were 0.02, 0.10, 0.04, 0.40, 0.02 ng, respectively. The limits of quantitation were 0.04, 0.20, 0.08, 0.80, 0.04 ng, separately. The RSDs of precision, reproducibility and stability (24 h) tests were no more than 3.5%. The recovery rates were 99.3%-100.75% (RSD=2.09%-3.17%). Conclusions The method established in this experiment is simple, rapid, sensitive and accurate. It can be used to simultaneously determine the contents of five components in Ganmaoling granules, and can be used for quality control of Ganmaoning granules. Key words: Quality control; Ganmaoning granules; UPLC-MS/MS; Adenosine; Chlorogenic acid; Caffeic acid; Baicalin; Wogonin

Active compounds present inRosmarinus officinalis leaves andScutellaria baicalensis root evaluated as new therapeutic agents for endometriosis.

Can carnosic acid, (CA) rosmarinic acid (RA) and wogonin (WG) inhibit the growth of cultured human endometrial stromal cells and endometriotic-like lesions induced in a BALB/c model of endometriosis? Primary stromal cell cultures were established from endometrial biopsies from women with endometriosis and controls. The human endometrial stromal cell line T-HESC was also used for in-vitro experiments. Endometriosis was surgically induced in BALB/c mice, which were randomly assigned to CA 2 mg/kg/day (n = 11); CA 20 mg/kg/day (n = 10); RA 1 mg/kg/day (n = 11); RA 3 mg/kg/day (n = 10); WG 20 mg/kg/day (n = 12); intraperitoneal vehicle control (n = 8) or oral vehicle control (n = 11). After surgery, CA and RA were administered intraperitoneally on days 14-28. WG was administered orally by intragastric gavage on days 14-26. CA, RA and WG significantly inhibited in-vitro cell proliferation in primary and T-HESC cell cultures (P < 0.05). CA and WG induced cell cycle arrest of T-HESC at the G2/M phase (P < 0.01). RA reduced intracellular ROS accumulation (P < 0.001), whereas WG increased it (P < 0.05). WG significantly inhibited oestrogen receptor alpha expression in T-HESC (P < 0.01). In-vivo, CA, RA and WG significantly reduced lesions size (P < 0.05). All compounds significantly decreased the percentage of cells in proliferation (P < 0.05) whereas RA and WG further increased the percentage of apoptotic cells (P < 0.05) in endometriotic-like lesions. The results are promising; further investigation of these compounds as new therapeutics is needed.

Self-assembled amphiphilic zein-lactoferrin micelles for tumor targeted co-delivery of rapamycin and wogonin to breast cancer

Protein-based micelles have shown significant potential for tumor-targeted delivery of anti-cancer drugs. In this light, self-assembled nanocarriers based on GRAS (Generally recognized as safe) amphiphilic protein co-polymers were synthesized via carbodiimide coupling reaction. The new nano-platform is composed of the following key components: (i) hydrophobic zein core to encapsulate the hydrophobic drugs rapamycin (RAP) and wogonin (WOG) with high encapsulation efficiency, (ii) hydrophilic lactoferrin (Lf) corona to enhance the tumor targeting, and prolong systemic circulation of the nanocarriers, and (iii) glutaraldehyde (GLA)-crosslinking to reduce the particle size and improve micellar stability. Zein-Lf micelles showed relatively rapid release of WOG followed by slower diffusion of RAP from zein core. This sequential release may aid in efflux pump inhibition by WOG thus sensitizing tumor cells to RAP action. Interestingly, these micelles showed good hemocompatibility as well as enhanced serum stability owing to the brush-like architecture of Lf shell. Moreover, this combined nano-delivery system maximized synergistic cytotoxicity of RAP and WOG in terms of tumor inhibition in MCF-7 breast cancer cells and Ehrlich ascites tumor animal model as a result of enhanced active targeting. Collectively, GLA-crosslinked zein-Lf micelles hold great promise for combined RAP/WOG delivery to breast cancer with reduced drug dose, minimized side effects and maximized anti-tumor efficacy.

Mechanism in the existent difference in form of wogonin/wogonoside between plasma and intestine/liver in rats.

Wogonin (WO) and its glucuronide, wogonoside (WG) exhibit various beneficial bioactivities that may have potential for the development of novel drugs. In this study, we determined their pharmacokinetic characteristics in rats after intragastric administration of WO and intraportal vein injection of WG. WG was the predominant form in the portal vein and body plasma, and in bile; WO was detected only in the small intestine and liver. WG is a substrate of the multidrug resistance-associated protein (MRP) 1, 2, 3, and 4, and organic anion-transporting polypeptide (OATP) 2B1 and OATP1B3. Metabolism studies indicated that WG formation and WO decrease had similar CLint values in rat intestine S9 (RIS9) and rat liver microsome (RLM), and that the hydrolysis rate of WG in RIS9 and rat liver S9 (RLS9) was fast. Thus, WG could be excreted into the intestinal tract by MRP2, and transported into mesenteric blood by MRP1, 3, and 4. OATP2B1 and OATP1B3 mediated the hepatic uptake of WG and MRPs mediated WG efflux to the bile and circulation. The high transport capability of MRPs for WG and the fast hydrolysis in the small intestine and liver may be responsible for the presence of WO in these tissues.