Probiotic cocktails accelerate baicalin metabolism in the ileum to modulate intestinal health in broiler chickens

Background Baicalin and probiotic cocktails are promising feed additives with broad application prospects.While probiotic cocktails are known to enhance intestinal health,the potential synergistic impact of combining baicalin with probiotic cocktails on the gut health of broiler chickens remains largely unexplored.Therefore,this study aims to investigate the influence of the combined administration of baicalin and probiotic cocktails on the composition of ileal and cecal microbiota in broiler chickens to elucidate the underlying mechanisms responsible for the healthpromoting effects.Results A total of 3201-day-old male Arbor Acres broilers were divided into 4 groups,each with 8 replicates of 10 chicks per replicate.Over a period of 42 d,the birds were fed a basal diet or the same diet supplemented with 37.5 g/t baicalin(BC),1,000 g/t probiotic cocktails(PC),or a combination of both BC(37.5 g/t)and PC(1,000 g/t).The results demonstrated that BC+PC exhibited positive synergistic effects,enhancing intestinal morphology,immune function,and barrier function.This was evidenced by increased VH/CD ratio,sIgA levels,and upregulated expression of occludin and claudin-1(P<0.05).16S rRNA analysis indicated that PC potentiated the effects of BC,particularly in the ileum,where BC+PC significantly increased theα-diversity of the ileal microbiota,altered itsβ-diversity,and increased the relative abundance of Flavonifractor(P<0.05),a flavonoid-metabolizing bacterium.Furthermore,Flavonifractor positively correlated with chicken ileum crypt depth(P<0.05).While BC+PC had a limited effect on cecal microbiota structure,the PC group had a very similar microbial composition to BC+PC,suggesting that the effect of PC at the distal end of the gut overshadowed those of BC.Conclusions We demonstrated the synergistic enhancement of gut health regulation in broiler chickens by combining baicalin and probiotic cocktails.Probiotic cocktails enhanced the effects of baicalin and accelerated its metabolism in the ileum,thereby influencing the ileal microbiota structure.This study elucidates the interaction mechanism between probiotic cocktails and plant extract additives within the host microbiota.These findings provide compelling evidence for the future development of feed additive combinations.

Baicalin:a prominent therapeutic agent against colorectal cancer

The occurrence and development of colorectal cancer involve multiple genes and pathways as a result of accumulated mutations at several sites that control growth and differentiation.Baicalin is a flavonoid extracted from Scutellaria baicalensis Georgi with antioxidant,anti-inflammatory and antiviral activities.Here,we discuss its possible clinical development and perspectives for future research.We also summarize the literature on colorectal cancer as well as the anticancer effect of Scutellaria baicalensis Georgi in the last 20 years.As research progresses,the therapeutic effect of baicalin in combating colorectal cancer has gradually been recognized.Its impact on colorectal cancer and anticancer mechanism,such as blocking the tumor cell cycle,inducing tumor cell apoptosis,preventing and treating tumor metastasis and anti-inflammatory activity,have become new hot topics in the study of antitumor agents in Chinese medicine.The present review surveys and summarizes studies of baicalin in anti-colorectal cancer treatment to enhance the understanding of its tumoricidal mechanism and to provide new therapeutic options for colorectal cancer.

Effect of Baicalin on TNBS-Induced Colonic Inflammatory Injury in Rats

Objective: The aim is to observe the protective effect of baicalin on trinitrobenzene sulfonic acid (TNBS)-induced experimental colitis in mice and explore its mechanism. Methods: The mice were divided into 4 groups: ethanol control group, TNBS model group, baicalin low-dose group and baicalin high-dose group. The model of experimental colitis in mice was induced by TNBS enema. After 2 hours of TNBS enema, baicalin was given by gavage, QD × 7D. The animals were sacrificed on the 8th day to observe the extent of colonic mucosal damage, and the Peroxidase activity, Malondialdehyde (MDA) and glutathione (GSH) contents were measured. Results: Compared with the TNBS model group, the body weight, gross injury score and histological changes were significantly improved;MPO enzyme activity and MDA content were significantly decreased in the low and high-dose baicalin groups;and the content of glutathione increased. Conclusion: Baicalin can alleviate TNBS-induced colitis in mice, and the mechanism is related to the antioxidation of baicalin.

Anti-bacterial mechanism of baicalin-tobramycin combination on carbapenem-resistant Pseudomonas aeruginosa

BACKGROUND Pseudomonas aeruginosa(P.aeruginosa)is an important cause of nosocomial infections,and contributes to high morbidity and mortality,especially in intensive care units.P.aeruginosa is considered a'critical'category bacterial pathogen by the World Health Organization to encourage an urgent need for research and development of new antibiotics against its infections.AIM To investigate the effectiveness of baicalin combined with tobramycin therapy as a potential treatment method for carbapenem-resistant P.aeruginosa(CRPA)infections.METHODS Polymerase chain reaction(PCR)and RT-PCR were used to detect the expression levels of drug-resistant genes(including VIM,IMP and OprD2)and biofilmrelated genes(including algD,pslA and lasR)in CRPA that confer resistance to tobramycin,baicalin and tobramycin combined with baicalin(0,1/8,1/4,1/2 and 1MIC).RESULTS There was a correlation between biofilm formation and the expression of biofilmrelated genes.In addition,VIM,IMP,OprD2,algD,pslA and lasR that confer biofilm production under different concentrations in CRPA were significantly correlated.The synergistic effect of baicalin combined with tobramycin was a significant down-regulation of VIM,IMP,algD,pslA and lasR.CONCLUSION Baicalin combined with tobramycin therapy can be an effective treatment method for patients with CRPA infection.

Hippocampus protection from apoptosis by Baicalin in a LiClpilocarpine-induced rat status epilepticus model through autophagy activation

BACKGROUND Autophagy is associated with hippocampal injury following status epilepticus(SE)and is considered a potential therapeutic mechanism.Baicalin,an emerging multitherapeutic drug,has shown neuroprotective effects in patients with nervous system diseases due to its antioxidant properties.AIM To investigate the potential role of autophagy in LiCl-pilocarpine-induced SE.METHODS The drugs were administered 30 min before SE.Nissl staining showed that Baicalin attenuated hippocampal injury and reduced neuronal death in the hippocampus.Western blotting and terminal deoxynucleotidyl transferase dUTP nick end labeling assay confirmed that Baicalin reversed the expression intensity of cleaved caspase-3 and apoptosis in hippocampal CA1 following SE.Furthermore,western blotting and immunofluorescence staining were used to measure the expression of autophagy markers(p62/SQSTM1,Beclin 1,and LC3)and apoptotic pathway markers(cleaved caspase-3 and Bcl-2).RESULTS Baicalin significantly upregulated autophagic activity and downregulated mitochondrial apoptotic pathway markers.Conversely,3-methyladenine,a commonly used autophagy inhibitor,was simultaneously administered to inhibit the Baicalin-induced autophagy,abrogating the protective effect of Baicalin on the mitochondrial apoptotic level.CONCLUSION We illustrated that Baicalin-induced activation of autophagy alleviates apoptotic death and protects the hippocampus of SE rats.

Effects of baicalin in CD4 + CD29 + T cell subsets of ulcerative colitis patients

AIM:To evaluate the role of baicalin in ulcerative colitis(UC) with regard to the CD4+CD29+ T helper cell,its surface markers and serum inflammatory cytokines.METHODS:Flow cytometry was used to detect the percentage of CD4+CD29+ cells in patients with UC.Real time polymerase chain reaction was used to detect expression of GATA-3,forkhead box P3,T-box expressed in T cells(T-bet),and retinoic acid-related orphan nuclear hormone receptor C(RORC).Western blotting was used to analyze expression of nuclear factor-κB(NF-κB) p65,phosphorylation of NF-κB(p-NF-κB) p65,STAT4,p-STAT4,STAT6 and p-STAT6.The concentrations of interferon-γ(IFN-γ),interleukin(IL)-4,IL-5,IL-6,IL-10 and TGF-β in serum were determined by ELISA assay.RESULTS:The percentages of CD4+CD29+ T cells were lower in treatment with 40 and 20 μmol/L baicalin than in the treatment of no baicalin.Treatment with 40 or 20 μmol/L baicalin significantly upregulated expression of IL-4,TGF-β1 and IL-10,increased p-STAT6/STAT6 ratio,but downregulated expression of IFN-γ,IL-5,IL-6,RORC,Foxp3 and T-bet,and decreased ratios of T-bet/GATA-3,p-STAT4/STAT4 and p-NF-κB/NF-κB compared to the treatment of no baicalin.CONCLUSION:The results indicate that baicalin regulates immune balance and relieves the ulcerative colitis-induced inflammation reaction by promoting proliferation of CD4+CD29+ cells and modulating immunosuppressive pathways.

Nerve protective effect of Baicalin on newborn HIBD rats

Objetive:To investigate the nerve protective effect and mechanism of baicalin on newborn rats with hypoxic ischemic brain damage(HIBD).Methods:A total of 64 SD newborn rats were randomlu divided into control group.model group.nerve growth factor group and baicalin group.with 16 in each group.Left carotid artery ligation method was adopted to establish the HIBD model except fou in control group,which was treatde with intraperitoneal injection of salin e10mL/kg for 3 d.After oxygen recovery on hypoxia ischemia rats.intraperitoneal injectionof salin 10mL/kg was adopted in model group for 3 d.Intraperitoneal injection of nerve growth factor injection50μg/kg per day was adopted in nerve growth factor group for 3 d:intraperitoneal injection of radix scutellariae 16mg/kg per day was adopted in baicalin group for 3 d after modeeling.Four rats of each group were sacrificed at Day 1,2,3,7 for microscopic observation of pathological morphological changes in brain tissus aften HE staining,S-P immunohistochemical method was used for observation of Fas and FasL expression in brain cells.Results:Neat structure of cells was observed in control group;edema cells in disordered arrangement was observed in model group,with some cells necrosis and cavity change;tissue injury in nerve growth factor group and baicalin group was significantly lighter than that in model group;Fas and FasL expression in model group,nerve growth factor group and baicalin group were significantiy higher than that in control group at different time points(P<0.05):Fas and FasL expression in nerve growth factor group and baicalin group were significantly lower than that in model group at different time points(P<0.05):There was no statistical diggerence of Fas,FasL expression at each time point between nerve growth factor group and baicalin group(P>0.05).Conclusions:Baicalin can reduce expression of Fas and FasL in HIBD rats,inhibit apoptosis of nerve cells,thus achieve the protective effect on HIBD rat nerves.

Pathological changes in multiple organs of rats with severe acute pancreatitis treated by baicalin and octreotide

BACKGROUND:Severe acute pancreatitis(SAP)features fatal pathogenetic conditions and high mortality rate.The study of SAP complicated with multiple organ injuries is of important significance.In this study,we explored the protective effect of baicalin on multiple organs of SAP rats and compared it with that of octreotide through light and electron microscopic observations of the pathological changes. METHODS:The improved Aho method was used to prepare SAP rat models.These rats were then randomly divided into a sham-operated group(n=45),a model control group(n=45), baicalin-treated group(n=45)and octreotide-treated group (n=45).Based on the difference in time points after operation, these groups were subdivided into 3,6 and 12 hour subgroups (n=15).At the corresponding time point after operation,the mortality rate of rats was recorded,and then the rats were humanely killed to take samples of multiple organs that were subsequently examined for pathological changes under light and electron microscopy. RESULTS:At 12 hours after operation,the mortality rate of rats in the baicalin-and octreotide-treated groups was lower than that in the model control group(P【0.05).Compared to the model control group,the pathological changes and pathological scores in the baicalin-and octreotide-treated groups were mitigated and relieved to varying degrees.The pathological changes under electron microscopy were also improved.CONCLUSIONS:Both baicalin and octreotide show good protective effects on multiple organs of SAP rats.Baicalin as a new drug has good prospects in the treatment of SAP.

Baicalin attenuates blood-spinal cord barrier disruption and apoptosis through PI3K/Akt signaling pathway after spinal cord injury

Baicalin is a natural active ingredient isolated from Scutellariae Radix that can cross the blood-brain barrier and exhibits neuroprotective effects on multiple central nervous system diseases.However,the mechanism behind the neuroprotective effects remains unclear.In this study,rat models of spinal cord injury were established using a modified Allen's impact method and then treated with intraperitoneal injection of Baicalin.The results revealed that Baicalin greatly increased the Basso,Beattie,Bresnahan Locomotor Rating Scale score,reduced blood-spinal cord barrier permeability,decreased the expression of Bax,Caspase-3,and nuclear factorκB,increased the expression of Bcl-2,and reduced neuronal apoptosis and pathological spinal cord injury.SH-SY5 Y cell models of excitotoxicity were established by application of 10 m M glutamate for 12 hours and then treated with 40μM Baicalin for 48 hours to investigate the mechanism of action of Baicalin.The results showed that Baicalin reversed tight junction protein expression tendencies(occludin and ZO-1)and apoptosis-related protein expression(Bax,Bcl-2,Caspase-3,and nuclear factor-κB),and also led to up-regulation of PI3 K and Akt phosphorylation.These effects on Bax,Bcl-2,and Caspase-3 were blocked by pretreatment with the PI3 K inhibitor LY294002.These findings suggest that Baicalin can inhibit bloodspinal cord barrier permeability after spinal cord injury and reduce neuronal apoptosis,possibly by activating the PI3 K/Akt signaling pathway.This study was approved by Animal Ethics Committee of Xi'an Jiaotong University on March 6,2014.

Baicalin treatment regulates hyperactivity of HPA axis and alters SIRT1 related inflammation in the hypothalamus in a model of depression

OBJECTIVE Hyperactivityof hypothalamic-pituitary-adrenal(HPA) axis is an important aetiological risk factor for the development of depression. Previous studies have demonstrated that the phenolic monomer baicalin has antidepressant-like effects and decreases serum corticosterone levels.However,the mechanism by which baicalin regulates hyperactivity of HPA axis remains unclear. This work aimed to investigate the effects of baicalin on hyperactivity of HPA axis using the olfactory bulbectomised(OBX) rat model of depression. METHODS Animals were anaesthetised with 10% chloral hydrate(3.3 mL·kg^(-1),ip). Using disinfected surgical equipment,the skull covering the olfactory bulbs was exposed by a midline incision. Two burr holes(2 mm diameter) were drilled 8 mm anterior to the bregma and 2 mm lateral to the midline. Both olfactory bulbs were aspirated and the holes filled with glass ionomer cement. The scalp was sutured closed. Sham-operated rats underwent every surgical procedure except the aspiration of the bulbs. The animals received penicillin(8×10~5U) intramuscularly(0.2 mL/300 g) once per day for 3 d post-surgery to prevent infection,and were subsequently housed alone in polypropylene cages. The experiments continued after 14 d of rehabilitation. The following groups were used for experiments: sham-operated(underwent surgical procedure without aspiratedolfactory bulbs and administration of vehicle only),OBX-model(underwent every surgical procedure and administration of vehicle only),OBX-amitriptyline treated(10 mg·kg^(-1)),and OBX-baicalin treatment(20 and 40 mg·kg^(-1)). Amitriptyline and baicalin were dissolved in physiological saline. RESULTS We examined how baicalin altered OBX-induced changes in serum glucocorticoid level as wel as inflammatory responses,sirtuin 1(SIRT1) expression,and p65 acetylation in the hypothalamus. Similar experiments were performed to analyse the effects of baicalin on lipopolysaccharide-induced inflammatory responses inhypothalamus. CONCLUSION Our results indicate that activation of the SIRT1 in the hypothalamus contributes to hyperactivity of HPA axis,which can be alleviated by baicalin.

Role of baicalin as a potential therapeutic agent in hepatobiliary and gastrointestinal disorders:A review

Baicalin is a natural bioactive compound derived from Scutellaria baicalensis,which is extensively used in traditional Chinese medicine.A literature survey demonstrated the broad spectrum of health benefits of baicalin such as antioxidant,anticancer,anti-inflammatory,antimicrobial,cardio-protective,hepatoprotective,renal protective,and neuroprotective properties.Baicalin is hydrolyzed to its metabolite baicalein by the action of gut microbiota,which is further reconverted to baicalin via phase 2 metabolism in the liver.Many studies have suggested that baicalin exhibits therapeutic potential against several types of hepatic disorders including hepatic fibrosis,xenobiotic-induced liver injury,fatty liver disease,viral hepatitis,cholestasis,ulcerative colitis,hepatocellular and colorectal cancer.During in vitro and in vivo examinations,it has been observed that baicalin showed a protective role against liver and gut-associated abnormalities by modifying several signaling pathways such as nuclear factor-kappa B,transforming growth factor beta 1/SMAD3,sirtuin 1,p38/mitogen-activated protein kinase/Janus kinase,and calcium/calmodulin-dependent protein kinase kinaseβ/adenosine monophosphate-activated protein kinase/acetyl-coenzyme A carboxylase pathways.Furthermore,baicalin also regulates the expression of fibrotic genes such as smooth muscle actin,connective tissue growth factor,β-catenin,and inflammatory cytokines such as interferon gamma,interleukin-6(IL-6),tumor necrosis factor-alpha,and IL-1β,and attenuates the production of apoptotic proteins such as caspase-3,caspase-9 and B-cell lymphoma 2.However,due to its low solubility and poor bioavailability,widespread therapeutic applications of baicalin still remain a challenge.This review summarized the hepatic and gastrointestinal protective attributes of baicalin with an emphasis on the molecular mechanisms that regulate the interaction of baicalin with the gut microbiota.

High-throughput RNA sequencing reveals the anti-inflammatory mechanism of baicalin on Propionibacterium acnes-induced acne in rabbits

Objective:Propionibacterium acnes(P.acnes)plays an important role in the development of acne,an inflammatory skin disease with a high-incidence.In this study,we used high-throughput RNA sequencing(RNA-seq)to reveal the anti-inflammatory mechanism of baicalin on P.acnes-induced acne in rabbits.Methods:The Kligman method was used to induce acne in the ears of New Zealand rabbits.The effect of baicalin on the acne model was evaluated by the number of acne lesions and hematoxylin and eosin(H&E)staining of acne tissues.Enzyme-linked immunoabsorbent assay was used to measure the protein expression levels of tumor necrosis factor a(TNFA),interleukin-1 b(IL1B),IL6,and IL8 in the serum of rabbits.RNA-seq was performed to investigate the mechanism of anti-inflammatory activities of baicalin on acne.Immunohistochemical analysis and Western blot were used to validate the expression levels of related proteins in acne tissues.Results:Baicalin treatment significantly reduced the number of acne lesions and lesions of the ear as well as levels of serum inflammatory cytokines.RNA-seq data showed that baicalin treatment globally suppressed inflammation,especially the TNF signaling pathway and Staphylococcus aureus infection pathway,in the rabbit acne model.Conclusion: Baicalin effectively ameliorates P. acnes-induced acne in rabbits by suppressingthe inflammatory response in rabbits.

Simultaneous determination of chlorogenic acid and baicalin in heat-clearing and detoxicating oral liquid by NIRS

The calibration model for simultaneous deter-mination of chlorogenic acid and baicalin in heat-clearing and detoxicating oral liquid was built by partial least squares and near infrared spectroscopy, and the method of spectral pre-treatment was discussed. Building model from calibration set obtained good results, and vali-dated by prediction. According to heat-clearing and detoxicating oral liquid from 30 batches of 6 factories, the correlation coefficient of chloro-genic acid and baicalin model are 0.9993 and 0.9923, The root mean square error of cross validation (RMSECV) are 0.467 and 0.480, and the standard Error of prediction (SEP) of chloro- genic acid and baicalin are 0.356 and 0.370 re-spectively. The correlation coefficients in pre-diction set are 0.9997 and 0.9969, prediction results are accurate and reliable. This method can be applied in rapid analysis of heat- clearing and detoxicating oral liquid, and it is fit for on-line detection and has a wide application prospect.

The role of baicalin on carbon tetrachloride induced liver fibrosis

The effect of the baicalin,a bio-active flavonoid extracted from Scutellaria baicalensis Georgi,on the carbon tetrachloride(CCl_(4))induced liver fibrosis was investigated.To compare the effect of baicalin on the liver fibrosis,five different groups of rats treated by 100,200,and 400 mg/kg baicalin were studied.Upon CCl_(4) treatment,the levels of procollagen type III,aspartate aminotransferase,aminotransferase,hyaluronic acid,and hydroxyproline were significantly increased,whereas the superoxide dismutase and glutathione peroxidase content were decreased.These changes in the biochemical parameters,which are associated with liver function,were significantly attenuated by the baicalin treatment,suggesting that baicalin can suppress the liver fibrosis induced by CCl_(4).Moreover,the histological staining analysis demonstrated that baicalin could effectively inhibit the degree of liver cell injury.The protein expression of AKT/JAK2/ERK in the serum were markedly increased by CCl_(4) but suppressed by the treatment of baicalin in a dose-dependent manner,implying that baicalin can attenuated cell apoptosis induced by CCl_(4).Overall,these results suggest that baicalin effectively protects hepatocytes from the CCl_(4) oxidative damage,likely due to the inhibition of free radical generation and cell apoptosis during the liver injury.

Baicalin provides protection against fluoxetine-induced hepatotoxicity by modulation of oxidative stress and inflammation

BACKGROUND Fluoxetine is one of the most widely prescribed anti-depressant drugs belonging to the category of selective serotonin reuptake inhibitors.Long-term fluoxetine treatment results in hepatotoxicity.Baicalin,a natural compound obtained from the Chinese herb Scutellaria baicalensis is known to have antioxidant,hepatoprotective and anti-inflammatory effects.However,the beneficial effects of baicalin against fluoxetine-induced hepatic damage have not previously been reported.AIM To evaluate the protective action of baicalin in fluoxetine-induced liver toxicity and inflammation.METHODS Male albino Wistar rats were divided into seven groups.Group 1 was the normal control.Oral fluoxetine was administered at 10 mg/kg body weight to groups 2,3,4 and 5.In addition,groups 3 and 4 were also co-administered oral baicalin(50 mg/kg and 100 mg/kg,respectively)while group 5 received silymarin(100 mg/kg),a standard hepatoprotective compound for comparison.Groups 6 and 7 were used as a positive control for baicalin(100 mg/kg)and silymarin(100 mg/kg),respectively.All treatments were carried out for 28 d.After sacrifice of the rats,biomarkers of oxidative stress[superoxide dismutase(SOD),catalase(CAT),reduced glutathione(GSH),glutathione-S-transferase(GST),advanced oxidation protein products(AOPP),malondialdehyde(MDA)],and liver injury[alanine transaminase(ALT),aspartate transaminase(AST),alkaline phosphatase(ALP),total protein,albumin,bilirubin]were studied in serum and tissue using standard protocols and diagnostic kits.Inflammatory markers[tumor necrosis factor(TNF-α),interleukin(IL)-6,IL-10 and interferon(IFN)-γ]in serum were evaluated using ELISA-based kits.The effect of baicalin on liver was also analyzed by histopathological examination of tissue sections.RESULTS Fluoxetine-treated rats showed elevated levels of the serum liver function markers(total bilirubin,ALT,AST,and ALP)and inflammatory markers(TNF-α,IL-6,IL-10 and IFN-γ),with a decline in total protein and albumin levels.Biochemical markers of oxidative stress such as SOD,CAT,GST,GSH,MDA and AOPP in the liver tissue homogenate were also altered indicating a surge in reactive oxygen species leading to oxidative damage.Histological examination of liver tissue also showed degeneration of hepatocytes.Concurrent administration of baicalin(50 and 100 mg/kg)restored the biomarkers of oxidative stress,inflammation and hepatic damage in serum as well as in liver tissues to near normal levels.CONCLUSION These findings suggested that long-term treatment with fluoxetine leads to oxidative stress via the formation of free radicals that consequently cause inflammation and liver damage.Concurrent treatment with baicalin alleviated fluoxetine-induced hepatotoxicity and liver injury by regulating oxidative stress and inflammation.

Synergistic use of geniposide and baicalin on BV2 cell activation damage caused by LPS

OBJECTIVE To explore the synergistic effect of baicalin and geniposide(BG)on BV2 cell activation damage caused by lipopolysaccharide(LPS).METHODS BV2 murine microglial cell line was cultured in vitro,LPS(final concentration 500 ng·m L-1)and various concentrationof Baicalin and Geniposide(BG)(final concentration12.5,25 and 50μg·m L-1)were added tointerven,the negative control was establised.MTT method was used to value the effect of LPS on the viability of BV2 cell line.The accumulated nitrite was assayed utilizing the Griess reaction method.RESULTS(1)Morphological observation:The common marphological of quesient microglia is circle,cell bodies smaller and synaptic slender.The enlargement of microglial cell bodies and an amoeboid morphology with retraction of extensions are generally induced by LPS.BG markedly suppressed the LPS-activated BV2 microglia morphological variations,meanwhile the dose-dependent was dramaticaly performed.(2)MTT test showed that LPS-stimulated BV2 cells viability was significantly decreased compared to the control group;compared to LPS treated cells,drug group(LPS+BG)effectively improves the LPS-stimulated BV2 cells viability.(3)The Griess reaction method indicated that LPS could obviously promoted the BV2 cells′NO generation contrasted to control group;while the drug group(LPS+BG)can effectively inhibited the generation of NO which activated by LPS.CONCLUSION The treatment group could significantly enhance survival rate of LPSstimulated BV2 cells,while,the level of NO was markedly decreased in BV2 induced by LPS.These findings suggest that combination of BG could attenuate BV2 microglial cells activation and injury which induced by LPS,possessed the capacity of neuroprotective.

EFFECTS OF BAICALEIN, WOGONIN, BAICALIN AND Na2MoO_4 ON N-NITROSATION REACTION

The in vitro effects of baicalein,wogonin,baicalin and Na2MoO4 on N-nitrosation reaction have been studied. The N-nitrosation reaction has been determined by the amount of dimethylnitrosamine（DMN） and diethylnitrosamine（DEN） produced using the UV-photclysis spectrophotometric and pyrolysis gas chromatography test. Baicalein,wogonin and Na2MoO4 showed varing extents of inhibition of the formation of DMN and DEN. Baicalin promoted the formation of DMN and DEN under the condition of simulating gastric Juices. It is also found that E. coli showed a remarkable promoting effect on the formation of DMN and DEN,but this promoting effect could be blocked to some extent by baicalein,wogonin,baicalin and Na2MoO4. Besides, Na2MoO4 and wogonin have shown synergic effect on the blocking of N-nitrosation reaction.

Baicalin inhibits cell growth and induces autophagy via AMPK/ULK1 activation in MDA-MB-231 cells

OBJECTIVE To investigate the effects and molecular mechanisms of Baicalin,a natural flavonoid compound derived from Scutellariabaicalensis Georgi,in the triple-negative human breast cancer cell line MDAMB-231.METHODS Cel s(1.0×105mL-1)were seeded in96-well plates,6-well plates or 25 cm2flasks.After overnight incubation,various concentrations of Baicalin were added to cells for another 48 h.Cell viability was measured using XTT Assay.Cell growth and migration was measured using colony formation assay and wound healing assay,autophagy-related proteins were observed using Western blotting analysis.RESULTS A dose-dependent decrease in cell viability was induced by Baicalin(IC50=48.6μg·m L-1).The colony-forming activity of MDA-MB-231 cells was significantly reduced by various concentrations of baicalin(25,50,100μg·m L-1)(85.2±12.7%,41.3±12.3%,19.6±6.6%).Wound healing assay showed that the recovery rateof baicalin-treated groups(25,50,100μg·m L-1)were significantly lower than those of the untreated group(88.3±15.1%,52.1±15.5%,28.3±9.6%).Western blot showed that the AMP-activated protein kinase and ULK1 was clearly up-regulated and activated by Baicalin(25,50,100μg·m L-1)in a dose-dependent manner,and the expression level of autophagic marker Beclin-1 and LC3A/B was also unregulated by the same treatment.CONCLUSION The study revealed that baicalin interferes with breast cancer growth by inducing autophagy,which at least in part through AMPK/ULK1 activation.

The protection effect of compatibility of baicalin and gardenoside on the blood-brain barrier damage

Aim To investigate the protection effect of the compatibility of baicalin and geniposide （7 ： 3 ） on blood-brain barrier （BBB） damage and the mechanism of down-regulating the expression of AQP-4 protein in cere- bral ischemia reperfusion injury （CIRI） rats. Method： 100 rats were divided into 5 groups： sham, CIRI model group, baicalin and geniposide （7 ： 3） （30 mg · kg^-1 ,60 mg · kg^-1） group, allyl chloride （0. 0021 ml· kg^-1）group. The model of CIRI made by improved suture method, Neural function defect, morphology and number of neurons in cerebral cortex was observed by Nissl staining; tested the contental change of P-gp and Na＋ , K＋-ATP enzymes of brain tissue, the contental change of S100β, Glucose, pyruvic acid and lactic acid of plasma by ELISA; the dry wet weight and Evans Blue （EB） tracing method served BBB permeabilitical changes; immunohis- tochemistry staining and semi-quantitation analysis were performed to detect the AQP-4 and GFAP in cerebra ische- mia; the expression of AQP-4 in cerebra hippocampus was determined by RT-qPCR and Western blot; pathological change was observed in brain issueby HE staining; observed the changes of brain tissue ultrastructure usingtrans- mission electron microscope with Lanthanum nitrate tracer ; monitored the size and location of the ischemia injury in brain regions with magnetic resonance imaging （MRI）. Results Compared with CIRI group, baicalin and genipo- side group can restore nerve function defect; increase the number of Nissl positive cells in cerebral cortex; weaken Na＋ ,K＋-ATP enzyme dynamic, reduce the content of S10013 Glucose and P-gp, reduce the content of water and volume EB in brain, elevated the7content of pyruvic acid and pyruvic acid; remarkable attenuation of AQP-4 and GFAP over-expression in the brain; remarkable attenuation of AQP-4 mRNA expression in hippocampus; The mor- phology is became clear, eased lumen of blood vessel compression deformation, loose organization, lessened cell volume and edema; Reduction of lanthanum particles into the blood vessels and the cells, reduce the vascular endo- thelial cell edema. Conclusion Baicalin and geniposide （7 ： 3 ） can reduce the permeability of blood brain barri- er, and has a protective effect on the brain edema induced by CIRI.

Protection of Compatibility of Saikosapon d and Baicalin on Carbon Tetrachloride Injured L-02 Cells Based on TLR4-NFκB Signaling Pathway

[Objectives] To study the protection of compatibility of Saikosapon d and Baicalin on carbon tetrachloride( CCl_4) injured L-02 cells. [Methods] Normal human hepatocyte cell line L-02 cells were cultured in vitro,and CCl_4 was used to induce hepatocellular injury. Interventions were carried out with Saikosaponin d and Baicalin at different dosage. The proliferation of L-02 cells in each group was determined by methylthiazolyl tetrazolium( MTT) assay; the levels of AST and ALT in the culture supernatants were detected by enzyme-linked immunosorbent assay( ELISA); the expressions of TLR4 and NFκBp65 proteins in each group were determined by immunohistochemistry.[Results] In the CCl_4 injured group,the proliferation of L-02 cells was significantly declined,the levels of AST and ALT in cell culture medium were significantly increased,and the expressions of TLR4 and NFκBp65 in L-02 cells were increased; after the intervention of Saikosaponin d and Baicalin,1. 75 μg/mL group and 1. 5 μg/mL group had an effect of promoting the proliferation of L-02 cells and could reduce the levels of AST and ALT in the cell culture medium,and TLR4 and NFκBp65 proteins in L-02 cells also had a certain inhibitory effect. [Conclusions] The compatibility of Saikosapon d and Baicalin has a certain protective effect on CCl_4 injured L-02 cells. The protection mechanism may be related with its down-regulating TLR4-NFκB signaling pathway and reducing the inflammation.