Flavonoid Baicalein Research Articles

Inhibitory Activity of Flavonoid Scaffolds on SARS-CoV-2 3CLpro: Insights from the Computational and Experimental Investigations.

The emergence of the COVID-19 situation has become a global issue due to the lack of effective antiviral drugs for treatment. Flavonoids are a class of plant secondary metabolites that have antiviral activity against SARS-CoV-2 through inhibition of the main protease (3CLpro). In this study, 22 flavonoids obtained from natural sources and semisynthetic approaches were investigated for their inhibitory activity against SARS-CoV-2 3CLpro, along with cytotoxicity on Vero cells. The protein-ligand interactions were examined using molecular dynamics simulation. Moreover, QSAR analysis was conducted to clarify the structural effects on bioactivity. Accordingly, the in vitro investigation demonstrated that four flavonoids, namely, tectochrysin (7), 6″,6″-dimethylchromeno-[2″,3″:7,8]-flavone (9), panduratin A (19), and genistein (20), showed higher protease inhibitory activity compared to the standard flavonoid baicalein. Finally, our finding suggests that genistein (20), an isoflavone discovered in Millettia brandisiana, has potential for further development as a SARS-CoV-2 3CLpro inhibitor.

Combining novel theoretical and experimental insights into the physical electrochemistry of gold and carbon-based electrodes and baicalein flavonoid sensing

Here, both the theoretical and experimental behavior of the flavonoid baicalein (BAI) was evaluated, considering the physical electrochemistry of carbon materials. For this purpose, experimental data based on cyclic voltammetry (CV) measurements were collected using the glassy carbon electrode (GCE) in comparison with a metallic gold electrode (GE). From a theoretical point of view, the focus was on modeling the BAI interaction with substrates through the use of graphene (its functionalizations) and a gold closed-packed slab model to understand the molecular adsorption process and verify important issues related to the analyte-substrate interaction at the atomistic level. The high-level quantum-mechanical calculations were based on density functional theory (DFT). From an analytical point of view, GCE and GE were tested for method development using differential pulse voltammetry (DPV). Regarding analytical purposes, limit of detection (LOD) values of 31.5 nmol L−1 and 68.1 nmol L−1 were estimated for GCE and GE, respectively. The standard addition method was explored with GE, while an external calibration curve approach was applied to overcome surface saturation on GCE. We found two elucidating points that may compromise electroanalysis: (i) in physisorption, the analyte adsorption occurs in parallel orientation to both substrate types, carbon- and gold-based ones, which compromises the useful electrode area; and (ii) in chemisorption, the presence of defects on the carbon-based electrode surface is responsible for a strong interaction that leads to BAI dissociation and electrode poisoning. Thus, the theoretical results support the mechanistic investigation, and novel insights into the electrochemical sensing of flavonoids are presented here.

Baicalin and lung diseases.

Studies focusing on natural products have been conducted worldwide, and the results suggest that their natural ingredients effectively treat a wide range of illnesses. Baicalin (BIA) is a glycoside derived from the flavonoid baicalein present in Scutellaria baicalensis of the Lamiaceae family. Interestingly, BIA has been shown to protect the lungs in several animal models used in numerous studies. Therefore, we fully analyzed the data of the studies that focused on BIA's lung protective function against various injuries and included them in this review. Interestingly, BIA exhibits promising effects against acute lung injury, lung fibrosis, pulmonary embolism, and lung remodelling associated with COPD, LPS, and paraquat insecticide. BAI exhibits anticancer activity against lung cancer. Additionally, BIA potently attenuates lung damage associated with infections. BIA primarily exerts its therapeutic effects by suppressing inflammation, oxidative stress immune response, and apoptosis pathways. Nrf2/HO-1, PI3K/Akt, NF-κB, STAT3, MAPKs, TLR4, and NLRP3 are important targets in the pulmonary therapeutic effects of BIA on different lung disease models. Consequently, we recommend using it in future potential clinical applications, its contribution to treatment guidelines, and translating its promising effects to clinical practice in lung diseases.

Novel Liposomal Formulation of Baicalein for the Treatment of Pancreatic Ductal Adenocarcinoma: Design, Characterization, and Evaluation.

Pancreatic cancer (PC) is one of the deadliest cancers so there is an urgent need to develop new drugs and therapies to treat it. Liposome-based formulations of naturally-derived bioactive compounds are promising anticancer candidates due to their potential for passive accumulation in tumor tissues, protection against payload degradation, and prevention of non-specific toxicity. We chose the naturally-derived flavonoid baicalein (BAI) due to its promising effect against pancreatic ductal adenocarcinoma (PDAC) and encapsulated it into a liposomal bilayer using the passive loading method, with an almost 90% efficiency. We performed a morphological and stability analysis of the obtained BAI liposomal formulation and evaluated its activity on two-dimensional and three-dimensional pancreatic cell models. As the result, we obtained a stable BAI-encapsulated liposomal suspension with a size of 100.9 nm ± 2.7 and homogeneity PDI = 0.124 ± 0.02, suitable for intravenous administration. Furthermore, this formulation showed high cytotoxic activity towards AsPC-1 and BxPC-3 PDAC cell lines (IC50 values ranging from 21 ± 3.6 µM to 27.6 ± 4.1 µM), with limited toxicity towards normal NHDF cells and a lack of hemolytic activity. Based on these results, this new BAI liposomal formulation is an excellent candidate for potential anti-PDAC therapy.

Comparative Evaluation of the Antiglycation and Anti-α-Glucosidase Activities of Baicalein, Baicalin (Baicalein 7-O-Glucuronide) and the Antidiabetic Drug Metformin

The discovery of new oral antidiabetic drugs remains a priority in medicine. This research aimed to evaluate the activity of the flavonoid baicalein and its natural glucuronide baicalin, compared to the antidiabetic drug metformin, as potential antiglycation, anti–radical, and anti-α–glucosidase agents, in order to assess their potential role in counteracting hyperglycemia-induced tissue damage. The study considered: (i) the BSA assay, to detect the formation of advanced glycation end products (AGEs), (ii) the GK peptide–ribose assay, which evaluates the cross–linking between the peptide and ribose, and (iii) the carbonyl content assay to detect the total carbonyl content, as a biomarker of tissue damage. In addition, to obtain a reliable picture of the antiglycation capacity of the investigated compounds, DPPH scavenging and oxygen radical absorbance capacity (ORAC) assays were performed. Furthermore, the anti–α–glucosidase activity of baicalein and baicalin was detected. Furthermore, to estimate cell permeability, preliminarily, the cytotoxicity of baicalein and baicalin was evaluated in HT–29 human colon adenocarcinoma cells using the MTT assay. Successively, the ability of the compounds to pass through the cytoplasmic membranes of HT–29 cells was detected as a permeability screen to predict in vivo absorption, showing that baicalein passes into cells even if it is quickly modified in various metabolites, being its main derivative baicalin. Otherwise, baicalin per se did not pass through cell membranes. Data show that baicalein is the most active compound in reducing glycation, α-glucosidase activity, and free radicals, while baicalin exhibited similar activities, but did not inhibit the enzyme α–glucosidase.

Flavonoid Baicalein Suppresses Oral Biofilms and Protects Enamel Hardness to Combat Dental Caries.

The objectives of this study were to investigate the effects of a novel method using flavonoids to inhibit Streptococcus mutans (S. mutans), Candida albicans (C. albicans) and dual-species biofilms and to protect enamel hardness in a biofilm-based caries model for the first time. Several flavonoids, including baicalein, naringenin and catechin, were tested. Gold-standard chlorhexidine (CHX) and untreated (UC) groups served as controls. Optimal concentrations were determined by cytotoxicity assay. Biofilm MTT, colony-forming-units (CFUs), biofilm biomass, lactic acid and polysaccharide production were evaluated. Real-time-polymerase-chain reaction (qRT-PCR) was used to determine gene expressions in biofilms. Demineralization of human enamel was induced via S. mutans-C. albicans biofilms, and enamel hardness was measured. Compared to CHX and UC groups, the baicalein group achieved the greatest reduction in S. mutans, C. albicans and S. mutans-C. albicans biofilms, yielding the least metabolic activity, polysaccharide synthesis and lactic acid production (p < 0.05). The biofilm CFU was decreased in baicalein group by 5 logs, 4 logs, 5 logs, for S. mutans, C. albicans and S. mutans-C. albicans biofilms, respectively, compared to UC group. When tested in a S. mutans-C. albicans in vitro caries model, the baicalein group substantially reduced enamel demineralization under biofilms, yielding an enamel hardness that was 2.75 times greater than that of UC group. Hence, the novel baicalein method is promising to inhibit dental caries by reducing biofilm formation and protecting enamel hardness.

Baicalein exhibits differential effects and mechanisms towards disruption of α-synuclein fibrils with different polymorphs

Parkinson's disease (PD) is the second most common neurodegenerative diseases with no cure yet and its major hallmark is α-synuclein fibrillary aggregates. The crucial role of α-synuclein aggregation in PD makes it an attractive target for potential disease-modifying therapies. Disaggregation of α-synuclein fibrils is considered as one of the promising therapeutic strategies to treat PD. The wild type (WT) and mutant α-synuclein fibrils exhibit different polymorphs and provide therapeutic targets for PD. Recent experiments reported that a flavonoid baicalein can disrupt WT α-synuclein fibrils. However, the underlying disruptive mechanism remains largely elusive, and whether BAC is capable of disrupting mutant α-synuclein fibrils is also unknown. Herein, we performed microsecond molecular dynamics simulations on cryo-EM-determined WT and two familial PD-associated mutant (E46K and H50Q) α-synuclein fibrils with and without baicalein. We find that baicalein destructs WT fibril by disrupting E46-K80 salt-bridge and β-sheets, and by remodeling the inter-protofilament interface. And baicalein can also damage E46K and H50Q mutant fibrils, but to different extents and via different mechanisms. The E46K fibril disruption is initiated from E61-K80 salt-bridge and N-terminal β-sheet, while the H50Q fibril disruption starts from the inter-protofilament interface and N-terminal β-sheet. These results reveal that disruptive effects and modes of baicalein on α-synuclein fibrils are polymorphism-dependent. This study suggests that baicalein may be a potential drug candidate to disrupt both WT and E46K/H50Q mutant α-synuclein fibrils and alleviate the pathological process of PD.

Affinity selection-mass spectrometry in the discovery of anti-SARS-CoV-2 compounds.

Small molecule therapeutic agents are needed to treat or prevent infections by severe acute respiratory syndrome coronavirus‐2 (SARS‐CoV‐2), which is the cause of the COVID‐19 pandemic. To expedite the discovery of lead compounds for development, assays have been developed based on affinity selection‐mass spectrometry (AS‐MS), which enables the rapid screening of mixtures such as combinatorial libraries and extracts of botanicals or other sources of natural products. AS‐MS assays have been used to find ligands to the SARS‐CoV‐2 spike protein for inhibition of cell entry as well as to the 3‐chymotrypsin‐like cysteine protease (3CLpro) and the RNA‐dependent RNA polymerase complex constituent Nsp9, which are targets for inhibition of viral replication. The AS‐MS approach of magnetic microbead affinity selection screening has been used to discover high‐affinity peptide ligands to the spike protein as well as the hemp cannabinoids cannabidiolic acid and cannabigerolic acid, which can prevent cell infection by SARS‐CoV‐2. Another AS‐MS method, native mass spectrometry, has been used to discover that the flavonoids baicalein, scutellarein, and ganhuangenin, can inhibit the SARS‐CoV‐2 protease 3CLpro. Native mass spectrometry has also been used to find an ent‐kaurane natural product, oridonin, that can bind to the viral protein Nsp9 and interfere with RNA replication. These natural lead compounds are under investigation for the development of therapeutic agents to prevent or treat SARS‐CoV‐2 infection.

Antioxidant, Analgesic, Antimicrobial and Molecular Docking Studies of the Leaves of Oroxylum indicum (L.) Kurz.

This study presents the findings of in vivo, in vitro and in silico investigations of the Oroxylum indicum (L.) Kurz leaf extracts. Various fractions namely chloroform (CF), ethyl acetate (EAF), n-hexane (HF) and aqueous fractions (AF) obtained from the crude extract were subjected to biological investigations. In DPPH based free radical scavenging assay, CF and MF fractions showed potent antioxidant activity with IC50 9.70 g/ml and 10.46 g/ml as compared to IC50 8.56g/ml for the standard ascorbic acid. The CF and HF fractions of O. indicum (L.) Kurz leaves exhibited mild antimicrobial activity. In analgesic activity screening by acetic acid induced writhing method, EAF showed significant writhing inhibition of 68% and 72%, at doses 150 mg/kg and 300 mg/kg, respectively. In tail immersion method, EAF also showed potent analgesic activity with reaction time of 1.816 ± 0.047 and 2.15 ± 0.041 min, after 30 minutes following oral doses of 150 mg/kg and 300 mg/kg, respectively. Molecular docking of previously isolated compounds from O. indicum (L.) Kurz leaf extracts were docked against cyclooxygenase-1 (COX-1) and cyclooxygenase -2 (COX-2) and revealed that the flavonoids baicalein and chrysin may be responsible for the analgesic activity. Lipinski’s rule of five was also calculated for the compounds to assess drug likeliness. Dhaka Univ. J. Pharm. Sci. 21(1): 85-94, 2022 (June)

Inhibition of Human UDP-Glucuronosyltransferases1A1-Mediated Bilirubin Glucuronidation by the Popular Flavonoids Baicalein, Baicalin, and Hyperoside Is Responsible for Herb (Shuang-Huang-Lian)-Induced Jaundice.

Bilirubin-related adverse drug reactions (ADRs) or malady (e.g., jaundice) induced by some herbs rich in certain flavonoids have been widely reported. However, the causes and mechanisms of the ADRs are not well understood. The aim of this paper was to explore the mechanism of Shuang-huang-lian (SHL) injections and its major constituents-induced jaundice via inhibiting human UDP-glucuronosyltransferases1A1 (hUGT1A1)-mediated bilirubin glucuronidation. The inhibitory effects of SHL and its major constituents in the herbal medicine, including baicalein (BAI), baicalin (BA), and hyperoside (HYP), on bilirubin glucuroBBREVInidation were investigated. This study indicated that the average formation rates of bilirubin glucuronides [i.e., mono-glucuronide 1 (BMG1), BMG2, and bilirubin diglucuronide] displayed significant differences (P < 0.05). Specifically, the formation of BMGs was favored regardless of whether an inhibitor was absent or present. SHL, BAI, BA, and HYP dose-dependently inhibit bilirubin glucuronidation, showing the IC50 values against total bilirubin glucuronidation were in the range of (7.69 ± 0.94)-(37.09 ± 2.03) μg/ml, (4.51 ± 0.27)-(20.84 ± 1.99) μM, (22.36 ± 5.74)-(41.35 ± 2.40) μM, and (15.16 ± 1.12)-(42.80 ± 2.63) μM for SHL, BAI, BA, and HYP, respectively. Both inhibition kinetics assays and molecular docking simulations suggested that SHL, BAI, BA, and HYP significantly inhibited hUGT1A1-mediated bilirubin glucuronidation via a mixed-type inhibition. Collectively, some naturally occurring flavonoids (BAI, BA, and HYP) in SHL have been identified as the inhibitors against hUGT1A1-mediated bilirubin glucuronidation, which well explains the bilirubin-related ADRs or malady triggered by SHL in clinical settings. SIGNIFICANCE STATEMENT: Herbal products and their components (e.g., flavonoids), which been widely used across the entire world, may cause liver injury. As a commonly used herbal products rich in flavonoids, SHL injections easily lead to symptoms of liver injury (e.g., jaundice) owing to significant inhibition of hUGT1A1-mediated bilirubin glucuronidation by its flavonoid components (i.e., baicalein, baicalin, and hyperoside). Herb-induced bilirubin-related ADRs and the associated clinical significance should be seriously considered.

Adenine overload induces ferroptosis in human primary proximal tubular epithelial cells

The pathogenesis of crystal nephropathy involves deposition of intratubular crystals, tubular obstruction and cell death. The deposition of 8-dihydroxyadenine (DHA) crystals within kidney tubules, for instance, is caused by a hereditary deficiency of adenine phosphoribosyl transferase in humans or adenine overload in preclinical models. However, the downstream pathobiological patterns of tubular cell attrition in adenine/DHA-induced nephropathy remain poorly understood. In this study, we investigated: (i) the modes of adenine-induced tubular cell death in an experimental rat model and in human primary proximal tubular epithelial cells (PTEC); and (ii) the therapeutic effect of the flavonoid baicalein as a novel cell death inhibitor. In a rat model of adenine diet-induced crystal nephropathy, significantly elevated levels of tubular iron deposition and lipid peroxidation (4-hydroxynonenal; 4-HNE) were detected. This phenotype is indicative of ferroptosis, a novel form of regulated necrosis. In cultures of human primary PTEC, adenine overload-induced significantly increased mitochondrial superoxide levels, mitochondrial depolarisation, DNA damage and necrotic cell death compared with untreated PTEC. Molecular interrogation of adenine-stimulated PTEC revealed a significant reduction in the lipid repair enzyme glutathione peroxidase 4 (GPX4) and the significant increase in 4-HNE compared with untreated PTEC, supporting the concept of ferroptotic cell death. Moreover, baicalein treatment inhibited ferroptosis in adenine-stimulated PTEC by selectively modulating the mitochondrial antioxidant enzyme superoxide dismutase 2 (SOD2) and thus, suppressing mitochondrial superoxide production and DNA damage. These data identify ferroptosis as the primary pattern of PTEC necrosis in adenine-induced nephropathy and establish baicalein as a potential therapeutic tool for the clinical management of ferroptosis-associated crystal nephropathies (e.g., DHA nephropathy, oxalate nephropathy).

The Flavonoid Baicalein Negatively Regulates Progesterone Target Genes in the Uterus in Vivo.

Baicalein is a flavonoid extracted from the root of Scutellaria baicalensis (Chinese skullcap) and is consumed as part of this botanical dietary supplement to reduce oxidative stress, pain, and inflammation. We previously reported that baicalein can also modify receptor signaling through the progesterone receptor (PR) and glucocorticoid receptor (GR) in vitro, which is interesting due to the well-established roles of both PR and GR in reducing inflammation. To understand the effects of baicalein on PR and GR signaling in vivo in the uterus, ovariectomized CD-1 mice were treated with DMSO, progesterone (P4), baicalein, P4 with baicalein, and P4 with RU486, a PR antagonist, for a week. The uteri were collected for histology and RNA sequencing. Our results showed that baicalein attenuated the antiproliferative effect of P4 on luminal epithelium as well as on the PR target genes HAND2 and ZBTB16. Baicalein did not change levels of PR or GR RNA or protein in the uterus. RNA sequencing data indicated that many transcripts significantly altered by baicalein were regulated in the opposite direction by P4. Similarly, a large portion of GO/KEGG terms and GSEA gene sets were altered in the opposite direction by baicalein as compared to P4 treatment. Treatment of baicalein did not change body weight, organ weight, or blood glucose level. In summary, baicalein functioned as a PR antagonist in vivo and therefore may oppose P4 action under certain conditions such as uterine hyperplasia, fibroids, and uterine cancers.

POS-377 BAICALEIN ATTENUATES ADENINE-INDUCED FERROPTOSIS IN HUMAN KIDNEY PRIMARY PROXIMAL TUBULAR EPITHELIAL CELLS (PTEC) VIA MAINTENANCE OF HEME OXYGENASE-1 EXPRESSION

Chronic kidney disease (CKD) is a global health burden, increasing in incidence worldwide. Adenine diet-induced CKD is a well-accepted pre-clinical model of CKD. In vitro, adenine-induced proximal tubular epithelial cell (PTEC) injury and loss are associated with increased oxidative stress, however little is known regarding the mechanisms of injury that cause PTEC loss. The aims of this study were (1) to investigate modes of cell death induced in human kidney primary PTEC by adenine, and (2) modulate any cell death with novel cell death inhibitors. Human kidney primary PTEC were cultured in the absence or presence of adenine and assessed for mitochondrial function, mitochondrial superoxides and altered cell viability by flow cytometry. Biomarkers of cell death pathways were analyzed by Western blotting. On identification of ferroptosis, rescue of PTEC by the plant-based anti-ferroptotic flavonoid baicalein was investigated. Adenine-treated PTEC displayed significantly reduced mitochondrial function and significantly increased mitochondrial superoxides and cell death compared with untreated PTEC. Further interrogation of cell death pathways revealed comparable expression of markers for apoptosis (activated caspase-3), necroptosis (pMLKL) and mitochondrial permeability transition-induced necrosis (PPIF) between untreated and adenine-treated PTEC. In contrast, the lipid repair enzyme GPX4, a biomarker for ferroptosis, was significantly reduced in adenine-treated PTEC, suggesting a pathway of ferroptototic cell death. Addition of the ferroptosis inhibitor baicalein restored expression of GPX4, maintained levels of antioxidant enzyme heme oxygenase-1 and reduced cell loss in adenine-treated PTEC. Our findings identify ferroptosis as a key human kidney PTEC cell death pathway in adenine-induced CKD. The anti-ferroptotic mechanisms of baicalein are now being explored for therapeutic translation in the clinical management of CKD.

Flavonoids' Effects on Caenorhabditis elegans' Longevity, Fat Accumulation, Stress Resistance and Gene Modulation Involve mTOR, SKN-1 and DAF-16.

Flavonoids are potential nutraceutical compounds present in diary food. They are considered health-promoting compounds and promising drugs for different diseases, such as neurological and inflammatory diseases, diabetes and cancer. Therefore, toxicological and mechanistic studies should be done to assert the biological effects and identify the molecular targets of these compounds. In this work we describe the effects of six structurally-related flavonoids—baicalein, chrysin, scutellarein, 6-hydroxyflavone, 6,7-dihydroxyflavone and 7,8-dihydroxyflavone—on Caenorhabditis elegans’ lifespan and stress resistance. The results showed that chrysin, 6-hydroxyflavone and baicalein prolonged C. elegans’ lifespan by up to 8.5%, 11.8% and 18.6%, respectively. The lifespan extensions caused by these flavonoids are dependent on different signaling pathways. The results suggested that chrysin’s effects are dependent on the insulin signaling pathway via DAF-16/FOXO. Baicalein and 6-hydroxyflavone’s effects are dependent on the SKN-1/Nfr2 pathway. In addition, microarray analysis showed that baicalein downregulates important age-related genes, such as mTOR and PARP.

Development of a Quantitative NMR Method for Direct and Simultaneous Determination of Four Flavonoids in Scutellaria baicalensis georgi Extracts

Introduction: Scutellaria Baicalensis Georgi (SBG), a Traditional Chinese Medicine (TCM) used for the treatment of antiviral therapy, contains many flavonoids. Materials and Methods: A simple, fast and accurate 1H quantitative nuclear magnetic resonance (1HqNMR) method was established for simultaneously determination of four flavonoids (Baicalin, Baicalein, Wogonin and Wogonoside) in SBG extract by using 3,4,5-trichloropyridine as an internal standard (IS). All the NMR determination work was performed at 308 K on an NMR sample tube inserted with a co-axial tube. The NMR sample tube was added with the sample (SBG extract and IS), the co-axial insert tube was added with D2O. Quantification of four flavonoids was carried out by calculating the relative peak area ratio of the selected proton signals from the target compounds and the IS. Results: The validated 1H-qNMR method was successfully used to determine the four flavonoids in 9 batches of SBG from different origins and the results were in good accordance with those obtained from HPLC. Conclusion: The established 1H-qNMR method could be used as a powerful tool for the quality control of SBG from different sources and had potential in the quantification of main components from other TCMs.

Flavonoids baicalein and kaempferol reduced inflammation in benign prostate hyperplasia patient-derived cells through regulating mitochondrial respiration and intracellular oxygen species

Flavonoids baicalein and kaempferol reduced inflammation in benign prostate hyperplasia patient-derived cells through regulating mitochondrial respiration and intracellular oxygen species

Enhanced topical permeability of natural flavonoid baicalein through nano liposomal gel: In vitro and in vivo investigation

Enhanced topical permeability of natural flavonoid baicalein through nano liposomal gel: In vitro and in vivo investigation

Evaluation of the post-treatment anti-inflammatory capacity of osteoarthritic chondrocytes: An invitro study using baicalein.

IntroductionTargeting inflammatory cascades is considered a promising way to prevent knee osteoarthritis (OA) progression. In terms of down-regulating the expression of inducible nitric oxide synthase (iNOS), interleukin (IL)-6, and matrix metalloproteinases (MMPs), pre-treatment with the flavonoid baicalein reportedly protects articular chondrocytes against the cytotoxicity of IL-1β. However, the benefits of post-treatment baicalein on osteoarthritic chondrocytes are not fully elucidated.MethodsIn this study, primary human chondrocytes were stimulated with IL-1β prior to baicalein application to evaluate the therapeutic effect of post-treatment.ResultsPost-treatment baicalein alleviated cell death and partially restored mitochondrial viability, while the senescence-associated secretory phenotype was not improved in IL-1β-stimulated chondrocytes. Post-treatment baicalein down-regulated the expressions of IL-1β, tumor necrosis factor-alpha, MMP-3, MMP-9, and MMP-13 mRNA as well as the protein production in stimulated cells. Even so, the levels of these factors were relative higher than those in un-treated chondrocytes. Moreover, iNOS, IL-6, IL-8, and COL1A1 expressions were consistently high, and IL-10 protein synthesis steadily increased in IL-1β-treated chondrocytes under baicalein treated status. Moreover, Western blot analyses showed that post-treatment baicalein suppressed nuclear factor kappa-light-chain-enhancer of activated B cells and p50 production while downstream cyclooxygenase-2 was still highly expressed.ConclusionBaicalein post-treatment to osteoarthritic chondrocytes had a minor benefit to the homeostasis of cartilaginous extracellular matrix.

Effect of the flavonoid baicalein as a feed additive on the growth performance, immunity, and antioxidant capacity of broiler chickens

Baicalein, the main flavonoid extracted from the root of Scutellaria baicalensis Georgi, has been demonstrated to exert multiple pharmacological effects, and thus could be utilized as a potential feed additive in broiler chickens. This study evaluated the effects of broiler chicken diet supplementation with baicalein on growth performance, immunity, and antioxidant activity at levels of 100 and 200mg/kg. No significant effect on average daily feed intake (P > 0.05) of broilers with diets supplemented with baicalein was found compared to those on the basal diet or butylated hydroxytoluene (BHT) during the 35-d feeding trial. The addition of baicalein to the basal diet significantly increased average body weight, body weight gain, average weight gain, and the feed conversion ratio of birds during 21 to 42 d and 7 to 42 d of age, respectively. The best numerical values for the overall growth performance were observed in broilers fed on diets containing 200mg/kg of baicalein. Baicalein supplementation significantly increased the ratio of CD3+/CD4+ and CD3±/CD8+, the concentration of IFN-γ, anti-IB antibody titer, and the spleen index compared with the control group (P < 0.05). Total cholesterol, the ratio of non-HDL-C/HDL-C, LDL-C/HDL-C, TC/HDL-C, triglycerides, and low-density lipoprotein cholesterol were significantly decreased after intake of baicalein compared with both the basal diet and the BHT-supplemented diet, whereas the SOD, GSH-Px, and CAT activity in the serum increased with the supplementation of baicalein. The T-AOC activity, T-SOD, and GSH-Px level in liver tissues was significantly increased by inclusion of baicalein, and intake of baicalein or BHT significantly decreased the malondialdehyde level found both in serum and meat tissue. Thus, the results obtained here indicate that the baicalein can be used as an effective natural feed additive in broiler chicken diets, and that 100 to 200mg/kg can be considered as the optimum dosage.

Baicalein inhibits fibronectin-induced epithelial\u2013mesenchymal transition by decreasing activation and upregulation of calpain-2

The extracellular matrix protein fibronectin (FN) facilitates tumorigenesis and the development of breast cancer. Inhibition of the FN-induced cellular response is a potential strategy for breast cancer treatment. In the present study, we investigated the effects of the flavonoid baicalein on FN-induced epithelial–mesenchymal transition (EMT) in MCF-10A breast epithelial cells and in a transgenic mouse MMTV-polyoma middle T antigen breast cancer model (MMTV-PyMT). Baicalein inhibited FN-induced migration, invasion, and F-actin remodeling. Baicalein also suppressed FN-induced downregulation of the epithelial markers E-cadherin and ZO-1 and upregulation of the mesenchymal markers N-cadherin, vimentin, and Snail. Further investigation revealed that calpain-2 was involved in baicalein suppression of FN-induced EMT. Baicalein significantly decreased FN-enhanced calpain-2 expression and activation by suppressing its plasma membrane localization, substrate cleavage, and degradation of its endogenous inhibitor calpastatin. Overexpression of calpain-2 in MCF-10A cells by gene transfection partially blocked the inhibitory effect of baicalein on FN-induced EMT changes. In addition, baicalein inhibited calpain-2 by decreasing FN-increased intracellular calcium ion levels and extracellular signal-regulated protein kinases activation. Baicalein significantly decreased tumor onset, growth, and pulmonary metastasis in a spontaneous breast cancer MMTV-PyMT mouse model. Baicalein also reduced the expression of FN, calpain-2, and vimentin, but increased E-cadherin expression in MMTV-PyMT mouse tumors. Overall, these results revealed that baicalein markedly inhibited FN-induced EMT by inhibiting calpain-2, thus providing novel insights into the pharmacological action and mechanism of baicalein. Baicalein may therefore possess therapeutic potential for the treatment of breast cancer though interfering with extracellular matrix–cancer cell interactions.