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 ...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.展开更多
Separation of baicalin from Scutellaria Baicalensis Georgi with polyamide was studied. The adsorption isotherm, kinetic equation and desorption law were investigated by static and dynamic adsorption methods. The resul...Separation of baicalin from Scutellaria Baicalensis Georgi with polyamide was studied. The adsorption isotherm, kinetic equation and desorption law were investigated by static and dynamic adsorption methods. The results show that the kinetic behavior is mainly controlled by the liquid film diffusion process and obeys the Boyd film diffusion equation. Equilibrium data for the adsorption of baicalin are correlated with Freundlich isotherm equation, i.e. q=3.8ce2.057, suggesting that the relative capacity of polyamide to baicalin is somewhat small. The desorption results indicate that the baicalin with mass fraction of 33.86% and the least impurities can be obtained by chromatography using 60% ethanol as the eluant at room temperature.展开更多
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 concent...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.展开更多
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 ...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.展开更多
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- ...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.展开更多
Baicalin(BA)is commonly used to treat inflammatory diseases and shows anti-inflammatory effects.The present study aimed to evaluate the analgesic and anti-inflammatory activities of BA both in vitro and in vivo.In ani...Baicalin(BA)is commonly used to treat inflammatory diseases and shows anti-inflammatory effects.The present study aimed to evaluate the analgesic and anti-inflammatory activities of BA both in vitro and in vivo.In animal models,acetic acid-induced writhing was used to assess analgesic activity.In addition,a variety of tests including xylene-induced ear edema test,minimum inhibitory concentration(MIC)assays and acetic acid-induced peritoneal capillary hyperpermeability test were used to evaluate antiinflammatory activity of BA.BA at 0.2 and 0.1 mg·mL-1 doses expressed analgesic as well as anti-inflammatory activities in mice.In acetic acid induced writhing test,BA applying three times,twice and once a day significantly inhibited the acetic acid-induced writhing response within 15 min by 7.80%(*p<0.05),7.50%(**p<0.01)and 6.25%(**p<0.01).In xylene-induced ear edema test,BA at 0.2,0.1 and 0.05 mg·mL-1 decreased ear edema by 45.50%(**p<0.01),15.20%(*p<0.05)and 9.10%(*p<0.05).In acetic acidinduced peritoneal capillary hyperpermeability test,BA exhibited significant inhibition(*p<0.05 versus control)of inflammation.In MIC assays,the MIC values of baicalin for S.aureus and Escherichia coli were 125 mg•mL-1,and the MIC values for the control bacteria ATCC25922 and ATCC25923 were 62.5 mg·mL-1.These findings suggested baicalin might contain analgesic and antiinflammatory agents which supported its use in traditional medicine.展开更多
OBJECTIVE Based on the methods of microdialysis,HPLC-MS/MS and gene chip tech.nology,the mechanism of Baicalin and Geniposide(BC/GP) against excitatory amino acid toxicity in ce.rebral ischemia was studied.This will p...OBJECTIVE Based on the methods of microdialysis,HPLC-MS/MS and gene chip tech.nology,the mechanism of Baicalin and Geniposide(BC/GP) against excitatory amino acid toxicity in ce.rebral ischemia was studied.This will provide guidance for the clinical application of BC/GP and the study of excitatory amino acid toxicity in cerebral ischemia.METHODS(1) Microdialysis technique and HPLC-MS/MS was performed to study the pharmacodynamics of BC/GP against cerebral ischemia.(1)18 SD rats with body weight of(280±20) g were randomly divided into control group,treatment groups with BC/CP at low dose,medium dose and high dose(equal to the dosage of crude drugs for 30 mg·kg^(-1),45 mg·kg^(-1) and 60 mg·kg^(-1) respectively).Rats in each group were given intragastric administration for seven days to establish cerebral ischemia model.Then,microdialysis probe was applied to collect cerebrospinal fluid from hippocampus before and after cerebral ischemia.(2) First,we established the HPLC-MS/MS method for measuring drugs and excitatory amino acids.Then we detected the microdi.alysis samples and observed their changes in animals.(2) The mechanism of BC/GP against excitatory toxicity of cerebral ischemia were observed at gene level by chip technique.(1) 16 SD rats with body weight of 240±20 g were randomly divided into sham group,model group,treatment group of BC(60 mg·kg^(-1)),treatment group of GP(60 mg·kg^(-1)) and treatment group of BC/GP(7:3)(60 mg·kg^(-1)).Rats in eachgroup were given intragastric administration for seven days to establish cerebral ischemia model.Then the rats were sacrificed,and the hippocampus were rapidly harvested and stored at-80℃ for further detection.(2) After the quality inspection of the hippocampal,the qualified samples were subjected to detect the levels of neurotransmitter receptor gene in the ischemic of rats by gene chip technology.Finally,the results were analyzed by the method of ΔΔCt.RESULTS(1) Only three compounds includ.ed GP,glutamic acid and aspartic acid were detected in microdialysis samples by HPLC-MS/MS.The concentration of GP increased and lasted for 120 min with a significant dose-dependent after cerebral ischemia.Compared with low dose group,the AUC(0-t),MRT(0-∞),Cmax and t1/2 z in high-dose group showed significant difference(P<0.01).Compared with the model group,the levels of glutamic acid and aspartic acid in the treatment groups decreased significantly,especially in the middle and high dose groups.(2)89 genes in the neurotransmitter receptor gene signaling pathway were detected by gene chip technol.ogy.There were 22 genes with |Fold Regulation| >1.5 in the model group,compared with the sham group.Five of the 22 genes showed statistically significant differences,including Grin2 c(2.9026),Chrna7(-1.5877),and Tacr2(-1.7695).Htr3 a(-1.8172) and Grm6(-2.3527).There were 5 genes with |Fold Regulation|>1.5 in the BC group,compared with the model group,Two of them exhibited statistically significant differences,including Brs3(1.797)and Grin2 c(-1.7979).There were 14 genes with |Fold Reg.ulation| >1.5 in the GP group,compared with the model group.Three of them displayed statistically significant differences,including Hcrtr2(-1.6584),Sctr(-3.8524) and Grin2 c(-4.8408).Compared with model group,the genes of |Fold Regulation| >1.5 in BC/GP(7:3) group are 5,and only one of them showed a significant differences.CONCLUSION(1) After administration of BC and GP,GP can cross the blood-brain barrier and reduce the release of excitatory amino acids in the hippocampus.(2) BC/GP can inhibit the interaction between excitatory amino acids and excitatory amino acid receptors and attenuate the toxicity of excitatory amino acids by down-regulating the expression of glutamic acid receptor Grin2 c gene.(3) BC/GP may exert their brain protection effect by reducing the release of excit.atory amino acids and inhibiting the expression of excitatory amino acid receptors.展开更多
文摘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.
基金Project(2006ABC014) supported by the Natural Science Foundation of Hubei Province, China
文摘Separation of baicalin from Scutellaria Baicalensis Georgi with polyamide was studied. The adsorption isotherm, kinetic equation and desorption law were investigated by static and dynamic adsorption methods. The results show that the kinetic behavior is mainly controlled by the liquid film diffusion process and obeys the Boyd film diffusion equation. Equilibrium data for the adsorption of baicalin are correlated with Freundlich isotherm equation, i.e. q=3.8ce2.057, suggesting that the relative capacity of polyamide to baicalin is somewhat small. The desorption results indicate that the baicalin with mass fraction of 33.86% and the least impurities can be obtained by chromatography using 60% ethanol as the eluant at room temperature.
基金The project suppored by National Natural Science Foundation of China(81473385)Shaanxi Province Education Department Project(13JS029)by Shaanxi Province Administration of Traditional Chinese Medicine(13-ZY016)
文摘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.
基金The project supported by National Natural Science Foundation of China(81403141)Nursery Project of Xiyuan Hospital,CACMS(XYKYMP2013-33)
文摘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.
文摘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.
基金Supported by the National Natural Science Foundation of China(31772801)Academic Backbone Project of Northeast Agricultural University(18XG23)。
文摘Baicalin(BA)is commonly used to treat inflammatory diseases and shows anti-inflammatory effects.The present study aimed to evaluate the analgesic and anti-inflammatory activities of BA both in vitro and in vivo.In animal models,acetic acid-induced writhing was used to assess analgesic activity.In addition,a variety of tests including xylene-induced ear edema test,minimum inhibitory concentration(MIC)assays and acetic acid-induced peritoneal capillary hyperpermeability test were used to evaluate antiinflammatory activity of BA.BA at 0.2 and 0.1 mg·mL-1 doses expressed analgesic as well as anti-inflammatory activities in mice.In acetic acid induced writhing test,BA applying three times,twice and once a day significantly inhibited the acetic acid-induced writhing response within 15 min by 7.80%(*p<0.05),7.50%(**p<0.01)and 6.25%(**p<0.01).In xylene-induced ear edema test,BA at 0.2,0.1 and 0.05 mg·mL-1 decreased ear edema by 45.50%(**p<0.01),15.20%(*p<0.05)and 9.10%(*p<0.05).In acetic acidinduced peritoneal capillary hyperpermeability test,BA exhibited significant inhibition(*p<0.05 versus control)of inflammation.In MIC assays,the MIC values of baicalin for S.aureus and Escherichia coli were 125 mg•mL-1,and the MIC values for the control bacteria ATCC25922 and ATCC25923 were 62.5 mg·mL-1.These findings suggested baicalin might contain analgesic and antiinflammatory agents which supported its use in traditional medicine.
基金supported by National Natural Science Foundation of China(81473385) Shaanxi provincial Natural Science Foundation of China(2017JZ027) Shaanxi Provincial Administration of Traditional Chinese Medicine(13-ZY016)
文摘OBJECTIVE Based on the methods of microdialysis,HPLC-MS/MS and gene chip tech.nology,the mechanism of Baicalin and Geniposide(BC/GP) against excitatory amino acid toxicity in ce.rebral ischemia was studied.This will provide guidance for the clinical application of BC/GP and the study of excitatory amino acid toxicity in cerebral ischemia.METHODS(1) Microdialysis technique and HPLC-MS/MS was performed to study the pharmacodynamics of BC/GP against cerebral ischemia.(1)18 SD rats with body weight of(280±20) g were randomly divided into control group,treatment groups with BC/CP at low dose,medium dose and high dose(equal to the dosage of crude drugs for 30 mg·kg^(-1),45 mg·kg^(-1) and 60 mg·kg^(-1) respectively).Rats in each group were given intragastric administration for seven days to establish cerebral ischemia model.Then,microdialysis probe was applied to collect cerebrospinal fluid from hippocampus before and after cerebral ischemia.(2) First,we established the HPLC-MS/MS method for measuring drugs and excitatory amino acids.Then we detected the microdi.alysis samples and observed their changes in animals.(2) The mechanism of BC/GP against excitatory toxicity of cerebral ischemia were observed at gene level by chip technique.(1) 16 SD rats with body weight of 240±20 g were randomly divided into sham group,model group,treatment group of BC(60 mg·kg^(-1)),treatment group of GP(60 mg·kg^(-1)) and treatment group of BC/GP(7:3)(60 mg·kg^(-1)).Rats in eachgroup were given intragastric administration for seven days to establish cerebral ischemia model.Then the rats were sacrificed,and the hippocampus were rapidly harvested and stored at-80℃ for further detection.(2) After the quality inspection of the hippocampal,the qualified samples were subjected to detect the levels of neurotransmitter receptor gene in the ischemic of rats by gene chip technology.Finally,the results were analyzed by the method of ΔΔCt.RESULTS(1) Only three compounds includ.ed GP,glutamic acid and aspartic acid were detected in microdialysis samples by HPLC-MS/MS.The concentration of GP increased and lasted for 120 min with a significant dose-dependent after cerebral ischemia.Compared with low dose group,the AUC(0-t),MRT(0-∞),Cmax and t1/2 z in high-dose group showed significant difference(P<0.01).Compared with the model group,the levels of glutamic acid and aspartic acid in the treatment groups decreased significantly,especially in the middle and high dose groups.(2)89 genes in the neurotransmitter receptor gene signaling pathway were detected by gene chip technol.ogy.There were 22 genes with |Fold Regulation| >1.5 in the model group,compared with the sham group.Five of the 22 genes showed statistically significant differences,including Grin2 c(2.9026),Chrna7(-1.5877),and Tacr2(-1.7695).Htr3 a(-1.8172) and Grm6(-2.3527).There were 5 genes with |Fold Regulation|>1.5 in the BC group,compared with the model group,Two of them exhibited statistically significant differences,including Brs3(1.797)and Grin2 c(-1.7979).There were 14 genes with |Fold Reg.ulation| >1.5 in the GP group,compared with the model group.Three of them displayed statistically significant differences,including Hcrtr2(-1.6584),Sctr(-3.8524) and Grin2 c(-4.8408).Compared with model group,the genes of |Fold Regulation| >1.5 in BC/GP(7:3) group are 5,and only one of them showed a significant differences.CONCLUSION(1) After administration of BC and GP,GP can cross the blood-brain barrier and reduce the release of excitatory amino acids in the hippocampus.(2) BC/GP can inhibit the interaction between excitatory amino acids and excitatory amino acid receptors and attenuate the toxicity of excitatory amino acids by down-regulating the expression of glutamic acid receptor Grin2 c gene.(3) BC/GP may exert their brain protection effect by reducing the release of excit.atory amino acids and inhibiting the expression of excitatory amino acid receptors.
