Baicalin and geniposide inhibit the development of atherosclerosis by increasing Wntl and inhibiting dickkopf-related protein-1 expression

Background Our previous study showed that the combined Chinese herbs containing scutellaria baicalensis georgi and gardenia jasminoids ellis inhibited atherosclerosis. In this study, we sought to determine if baicalin and geniposide could inhibit atherosclerosis through Wntl and dickkopf-related protein-1 （DKK1）. Methods The wild-type and ApoE-/- mice were treated with baicalin, geniposide, and baicalin plus geniposide daily by gavage for 12 weeks. Blood lipid levels were measured with an automatic biochemistry analyzer. Aortic atherosclerotic lesion areas were analyzed with Image-ProPlus software. The mRNA and protein expression of DKK1, Wntt and nuclear factor-r,B （NF-κB） were measured with RT-PCR and Westem Blot. Serum levels of interleukin-12 （IL-12） were quantified with ELISA. Results The baicalin or geniposide monotherapy as well as combination therapy inhibited the development of atherosclerotic lesions, increased Wntl and decreased DKKI expression and elevated the ratio of Wntl/DKK1 compared with high-lipid diet group. However, only baicalin or geniposide monotherapy decreased NF-κB expression. Moreover, baicalin and geniposide monoor combination therapy lowered IL-12 levels. Geniposide reduced both serum total cholesterol and low density lipoprotein levels, while baicalin either alone or in combination with geniposide did not affect serum lipid levels. In human, umbilical vein endothelial ceils stimulated by oxidized low density lipoprotein, baicalin and geniposide also increased Wntl and decreased DKK1 expression and elevated the ratio of Wntl/DKK1. Condusions Baicalin and geniposide exert inflammation-regulatory effects and may prevent atherosclerotic lesions through enhancing Wntl and inhibit- ing DKK1 expression.

Idiopathic mesenteric phlebosclerosis associated with long-term oral intake of geniposide

BACKGROUND Idiopathic mesenteric phlebosclerosis(IMP)is a rare disease,and its etiology and risk factors remain uncertain.AIM To investigate the possible influence of Chinese herbal liquid containing geniposide on IMP.METHODS The detailed formula of herbal liquid prescriptions of all patients was studied,and the herbal ingredients were compared to identify the toxic agent as a possible etiological factor.Abdominal computed tomography(CT)and colonoscopy images were reviewed to determine the extent and severity of mesenteric phlebosclerosis and the presence of findings regarding colitis.The disease CT score was determined by the distribution of mesenteric vein calcification and colon wall thickening on CT images.The drinking index of medicinal liquor was calculated from the daily quantity and drinking years of Chinese medicinal liquor.Subsequently,Spearman’s correlation analysis was conducted to evaluate the correlation between the drinking index and the CT disease score.RESULTS The mean age of the 8 enrolled patients was 75.7 years and male predominance was found(all 8 patients were men).The patients had histories of 5-40 years of oral Chinese herbal liquids containing geniposide and exhibited typical imaging characteristics(e.g.,threadlike calcifications along the colonic and mesenteric vessels or associated with a thickened colonic wall in CT images).Calcifications were confined to the right-side mesenteric vein in 6 of the 8 patients(75%)and involved the left-side mesenteric vein of 2 cases(25%)and the calcifications extended to the mesorectum in 1 of them.The thickening of colon wall mainly occurred in the right colon and the transverse colon.The median disease CT score was 4.88(n=7)and the median drinking index was 5680(n=7).After Spearman’s correlation analysis,the median CT score of the disease showed a significant positive correlation with the median drinking index(r=0.842,P<0.05).CONCLUSION Long-term oral intake of Chinese herbal liquid containing geniposide may play a role in the pathogenesis of IMP.

Geniposide, the component of the Chinese herbal formula Tongluojiunao, protects amyloid-β peptide(1–42)-mediated death of hippocampal neurons via the non-classical estrogen signaling pathway

Tongluojiunao （TLJN） is an herbal medicine consisting of two main components, geniposide and ginsenoside Rg1. TLJN has been shown to protect primary cultured hippocampal neurons. How-ever, its mechanism of action remains unclear. In the present study, primary cultured hippocampal neurons treated with Aβ1-42 （10 μmol/L） signiifcantly increased the release of lactate dehydroge-nase, which was markedly reduced by TLJN （2 μL/mL）, speciifcally by the component geniposide （26 μmol/L）, but not ginsenoside Rg1 （2.5 μmol/L）. hTe estrogen receptor inhibitor, ICI182780 （1 μmol/L）, did not block TLJN-or geniposide-mediated decrease of lactate dehydrogenase under Aβ1-42-exposed conditions. However, the phosphatidyl inositol 3-kinase or mitogen-activated protein kinase pathway inhibitor, LY294002 （50 μmol/L） or U0126 （10 μmol/L）, respectively blo cked the decrease of lactate dehydrogenase mediated by TLJN or geniposide. hTerefore, these results suggest that the non-classical estrogen pathway （i.e., phosphatidyl inositol 3-kinase or mitogen-activated protein kinase） is involved in the neuroprotective effect of TLJN, speciifcally its component, geniposide, against Aβ1-42-mediated cell death in primary cultured hippocampal neurons.

Geniposide protects human neuroblastoma SH-SY5Y cells against corticosterone-induced injury

In vitro cultured human neuroblastoma SH-SY5Y cells were pretreated with 50 or 5 ug/mL geniposide for 12 hours and exposed to 400 umol/L corticosterone. Corticosterone exposure in cultures not pretreated with geniposide resulted in inhibited cell growth, reduced cell survival, and increased P53 and P21 protein expression. However, in geniposide pretreated SH-SY5Y cells, cell viability and the number of cells in the G2 phase of the cell cycle were significantly increased, P21 and P53 protein expression was reduced, and cell apoptosis was inhibited following corticosterone exposure. These results indicate that geniposide can protect SH-SY5Y cells against high-dose corticosterone-induced injury.

Simultaneous determination of geniposidic acid. chlorogenic acid and geniposide in eucommia by HPLC

A high performance liquid chromatography (HPLC) method was established for simultaneous determina-tion of geniposidic acid, chlorogenic acid and geniposide in eucommia. Detection at 240 nm with a reversed-phasecolumn, CH3OH volume fraction, acidic additive and pH value of mobile phase were studied for their effects on theseparability of the compounds. The most suitable separation was obtained with isocratic gradient elution systemusing CH3OH-H2O-H3 PO4 (12.00: 87.96: 0.04, volume ratio) at a flow-rate of 1.0 mL/min. Under the experi-mental conditions, the capacity factors of three compounds are in 3-13. The sample is separated rightly. Theanalysis time is 30 min and the retention time of genfposidic acid, chlorogenic acid and geniposide are 6. 7 min,10.5 min and 21 min, respectively.

Effects of Pulse Ultrasound on Adsorption of Geniposide on Resin 1300 in a Fixed Bed

The effects of pulse ultrasound with different pulse parameters on the breakthrough curves of Geniposide on Resin 1300 were studied. The mass transfer model describing the adsorption process was constructed. Adsorption capability and the overall mass-transfer coefficient were obtained by fitting the constructed mass-transfer model and the experimental data. The effects of pulse ultrasound on adsorption of Geniposide on Resin1300 in a fixed bed were studied and compared. Amount of Geniposide adsorbed on Resin 1300 in the presence of ultrasound is lower than that in the absence of ultrasound, but the mass-transfer rate with ultrasonic irradiation is higher than that without ultrasound. Furthermore, mass transfer rate is enhanced by pulse modulation. In the conditions studied, the adsorption equilibrium constant decreases with increasing ultrasonic power, while the overall mass-transfer co-efficient increases. With increasing pulse duty ratio, adsorption equilibrium constant decreases initially, reaches a minimum when pulse duty ratio is 0.5, and then increases. On the contrary, the overall mass-transfer coefficient in-creases initially and reaches a maximum when pulse ratio is 0.5, and then decreases. Effects of pulse period on ad-sorption equilibrium and mass transfer rate reached the peak at pulse period of 28.6 ms.

Geniposide prevents rotenone-induced apoptosis in primary cultured neurons

Geniposide, a monomer extracted from gardenia and widely used in Chinese medicine, is a novel agonist at the glucagon-like peptide-1 receptor. This receptor is involved in neuroprotection. In the present study, we sought to identify an anti-apoptotic mechanism for the treatment of neurodegenerative diseases. Primary cultured neurons were treated with different concentrations of rotenone for 48 hours. Morphological observation, cell counting kit-8 assay, lactate dehydrogenase detection and western blot assay demonstrated that 0.5 n M rotenone increased lactate dehydrogenase release, decreased the expression of procaspase-3 and Bcl-2, and increased cleaved caspase-3 expression in normal neurons. All these effects were prevented by geniposide. Our results indicate that geniposide diminished rotenone-induced injury in primary neurons by suppressing apoptosis. This may be one of the molecular mechanisms underlying the efficacy of geniposide in the treatment of neurodegenerative diseases.

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.

Investigation of paeonol-geniposide on acute alcoholic liver injury based on uniform design method

Objective:To explore the optimal ratio and compatibility effect of paeonol-geniposide combination on acute alcoholic liver injury by uniform design.Methods:Lieber-DeCarli alcoholic liquid feed was used to induce acute alcoholic liver injury in mice.Uniform design was used to select the best dosage combination of paeonol and geniposide,and the related indexes of liver injury and oxidative stress were detected by kit.Serum inflammatory factors were detected by ELISA,and the expressions of p38 MAPK,JNK and NF-κB P65 related proteins in liver were detected by Western-blot.Results:The regression equation suggested that paeonol:geniposide=220:20 was the best ratio of paeonol and geniposide to resist alcoholic liver injury.Compared with the model group,the liver injury indexes and oxidation products of the paeonol+geniposide group decreased significantly,the antioxidant activity of liver tissue increased significantly,and the expression levels of p-p38 MAPK,p-JNK and NF-κB P65 protein decreased significantly.Conclusion:The optimal dosage of paeonolgeniposide was effectively optimized by uniform design and pharmacodynamic analysis.The combination of the two drugs could reduce the alcoholic liver injury by reducing the oxidative stress injury and inflammatory response in the liver tissue of mice,and its effect might be related to the targeting of p38 MAPK/JNK/NF-κB channel.

ERβ modulation and non-modulation of ERα by administration of geniposide and panax notoginseng saponins in SH-SY5Y cells

Objective:To illustrate the effect of geniposide (GP) and panax notoginseng saponins (PNS) on estrogen receptors (ER) including ERα and ERβ within the cytoplasm and nucleus of SH-SY5Y cells.Methods:Immunofluorescence was used to observe the distribution of ERα and ERβ in cytoplasm and nucleus,but Western blot was only for ERβ detection.q-PCR was applied to detect NR3C1,S100A6 and LGALS1downstream mRNA gene expression levels of ER.Results:Through analyzing fluorescence intensity under the administration of GP and PNS in SHSY5Y cells,we found that the distribution of ERα has not been affected.We also discovered that GP and/or PNS significantly stimulated the transportation of ERβ into the nucleus in a timedependent manner (all P <.001).When SH-SY5Y cells were treated with supplements of GP,PNS,GP + PNS at 15 minutes,30 minutes and 45 minutes,the distribution of ERβ in the nucleus significantly increased compared with that in control group (all P <.001).Evidently,treatment with GP,PNS,GP + PNS was able to significantly increase the levels of ERβ protein within the nucleus compared with control group at both 30 minutes and 45 minutes intervals (all P <.001).Furthermore,GP and PNS showed signs of activating to NR3C1 and LGALS1,two genes downstream of ER.It is possible that the 5100A6 gene mainly encoded the downstream gene in ERα's signaling pathway,which was not affected after the treatment of GP and/or PNS.Conclusion:The distribution and expression of ERβ has been modulated under the administration of GP + PNS within the SH-SY5Y cells,whereas ERα has not.GP and PNS in combination may play an estrogenic-like effect with selectivity on ERβ modulation.

Development of a method for the quantitative determination of geniposide in rat plasma by liquid chromatography-tandem mass spectrometry with positive/negative ion-switching electrospray ionization and its application in a pharmacokinetic study in rats

Geniposide is a major bioactive constituent isolated from Gardeniajasminoides Ellis. To evaluate the pharmacokinetics of geniposide in pre-clinical studies, a rapid and specific liquid chromatography-tandem mass spectrometry （LC-MS/MS） method was developed and validated. After simple protein precipitation, geniposide was analyzed on a DiamonsilR C18 column with a mobile phase of 10 mM ammonium acetate and methanol （20：80, v/v） at a flow rate of 0.6 mL/min. Detection was performed in ＂Truncated＂ multiple-reaction monitoring （MRM） mode with positive electrospray ionization （ESI） at m/z 411→411 for geniposide, and MRM mode with negative ESI ionization at m/z 415→295 for puerarin （internal standard, IS）. Linearity was established in the concentration range from 10.0 to 5000 ng/mL. The extraction recoveries ranged from 84.8% to 90.5% at concentrations of 10.0, 500 and 4.5x 103 ng/mL. The lower limit of quantification （LLOQ） was 10.0 ng/mL with 50 ~tL plasma. The validated method was successfully applied to the pharmacokinetic study of geniposide in rats at a dose of 200 mg/kg by oral administration.

The herbal compound geniposide rescues formaldehyde-induced apoptosis in N2a neuroblastoma cells

The herbal medicine Tong Luo Jiu Nao （TLJN） contains geniposide （GP） and ginsenoside Rgl at a molar ratio of i0：1. Rgl is the major component of another herbal medicine, panax notoginseng saponin （PNS）. TLJN has been shown to strengthen brain function in humans, and in animals it improves learning and memory. We have previously shown that TLJN reduces amyloi- dogenic processing in Alzheimer＇s disease （AD） mouse models. Together this suggests TLJN may be a potential treatment for patients with dementia. Because chronic damage of the central nervous system by formaldehyde （FA） has been presented as a risk factor for age-associated cognitive dysfunction, in the present study we investigated the protective effect of both TLJN and GP in neuron-like cells exposed to FA. FA-exposed murine N2a neuroblastoma cells were incubated with TLJN, its main in- gredient GP, as well as PNS, to measure cell viability and morphology, the rate of apoptosis and expression of genes encoding Akt, FOXO3, Bcl2 and p53. The CCK-8 assay, cytoskeletal staining and flow cytometry were used to test cell viability, mor- phology and apoptosis, respectively. Fluorescent quantitative real-time PCR （qRT-PCR） was used to monitor changes in gene expression, and HPLC to determine the rate of FA clearance. Treatment of N2a cells with 0.09 mmol L-1 FA for 24 h signifi- cantly reduced cell viability, changed cell morphology and promoted apoptosis. Both TLJN and GP conferred neuroprotection to FA-treated N2a cells, whereas PNS, which had to be used at lower concentrations because of its toxicity, did not. Our data demonstrate that TLJN can rescue neuronal damage caused by FA and that its main ingredient, GP, has a major role in this ef- ficacy. This presents purified GP as a drug or lead compound for the treatment of AD.

Geniposide inhibits CoCl2-induced PC12 cells death via the mitochondrial pathway

Background A number of studies have shown that oxidative stress and mitochondrial involvement are major triggering factors in the development of neurodegenerative diseases. Cobalt chloride （CoCl2）-induced cell death in PC12 cells may serve a simple and convenient in vitro model of hypoxia-induced neuronal cytotoxicity. To explore the effect of geniposide on COCl2 which induced cytotoxicity and mitochondrial function in rat pheochromocytoma PC12 cells, we analyzed the influence of geniposide on the expression of apoptosis-related proteins. Methods PC12 cells and RNAi PC12 cells were treated with 0, 12.5, 25, 50, 100 umol/L geniposide for 12 hours and then exposure to 400 umol/L COCI2 for 12 hours. Cell viability, cell morphology, and expression of Bcl-2, Bax, P53 and caspase-9 were determined using Western blotting. Results Pretreatment with geniposide markedly improved the cells viability and morphology, decreased the expression of Bax, P53 and caspase-9, and increased the expression of Bcl-2 in PC12 cells challenged by CoCl2. However, in the RNAi PC12 cells, geniposide had no significant effect on the expression of these proteins. Conclusion Geniposide protects PC12 cells from CoOl2 involved in mitochondrial mediated apoptosis, and GLP-1R might play a critical role in the neuroprotection of geniposide in PC12 cells.

Enhancing effect of natural borneol on the absorption of geniposide in rat via intranasal administration

Both geniposide （Ge） and natural borneol （NB） are bioactive substances derived from traditional Chinese herbs. The effect of NB on the pharmacokinetics of Ge in rat via intranasal administration was investigated. The concentrations of Ge in plasma were determined by reversed-phase high-performance liquid chromatography （HPLC） after intranasal administration of Ge （4 mg/kg） alone and combined with different doses （0.08, 0.8, and 8 mg/kg） of NB. The intravenous administration was given as a reference （4 mg/kg of Ge and 8 mg/kg of NB）. Compared with the intravenous administration, the absolute bioavailability of Ge was 76.14% through intranasal administration combined with NB. Compared with the intranasal administration of Ge alone, Ge could be absorbed rapidly in the nasal cavity combined with NB; the peak time of Ge in the plasma became shorter （3-5 min vs. 40 min）; the peak concentration became higher （1.32-4.25 IJg/ml vs. 0.67 ug/ml）; and, the relative bioavailability of Ge combined with NB was 90.3%-237.8%. The enhancing effect was attenuated as the dose of NB decreased. The results indicated that NB can accelerate the absorption of Ge dose-dependently in the nasal cavity.

Combination of Geniposide and Eleutheroside B Exerts Antidepressant-like Effect on Lipopolysaccharide-Induced Depression Mice Model

Objective To study the antidepressant-like effect and action mechanism of geniposide and eleutheroside B combination treatment on the lipopolysaccharide(LPS)-induced depression mice model.Methods Depression mice model was established by lipopolysaccharide(LPS)injection.Totally 48 mice were randomly divided into 6 groups(8 rats per group)according to a random number table,including normal,model,fluoxetine(20 mg/kg),geniposide(100 mg/kg)+eleutheroside B(100 mg/kg),geniposide+eleutheroside B+WAY 100635(0.03 mg/kg),geniposide+eleutheroside B+N-methyl-D-aspartic acid receptor(NMDA,75 mg/kg)groups,respectively.After continuous administration for 10 days,autonomic activity tests after 30 min of administration were performed on the 10th day.On the 11th day,except for the normal group,the mice in the other groups were intraperitoneally injected with LPS(1 mg/kg),and the behavioral tests were performed 4 h later.Enzyme linked immunosorbent assay was used to detect tumor necrosis factor alpha(TNF-α)and interleukin-1β(IL-1β)levels in mice serum.The mRNA expression of indoleamine 2,3-dioxygenase(IDO)and nuclear transcription factor(NF-κB)were detected by real-time quantitative polymerase chain reaction.Western-blot analysis was used to detect IDO and NF-κB protein expressions in hippocampus tissue.Results Compared with the normal group,a single administration of LPS increased the immobility time in the forced swimming test(FST)and tail suspension test(TST,P<0.01),without affecting autonomous activity.Compared with the model group,fluoxetine and geniposide+eleutheroside B administration significantly improved the immobility time of depressed mice in the FST and TST,decreased serum IL-1βcontent,inhibited the expression levels of NF-κB gene and protein in hippocampus tissues(P<0.05 or P<0.01).Compared with the model group,geniposide+eleutheroside B treatment significantly reduced serum TNF-αcontent and inhibited IDO mRNA and protein expressions in hippocampus(P<0.05 or P<0.01).In addition,NMDA partly prevented the inhibition of IDO mRNA expression by geniposide+eleutheroside B;NMDA and WAY-100635 also partly prevented the reduction of IL-1ßcontent induced by geniposide+eleutheroside B treatment(P<0.05 or P<0.01).Conclusions The combination of geniposide and eleutheroside B showed a certain antidepression-like effect.Its main mechanism of action may be contributed to inhibiting the activation of NF-κB,decreasing the proinflammatory cytokines such as TNF-α,IL-1β,and inhibiting in the neuroinflammatory reaction.Additionally,it also affects tryptophan metabolism,reduces the expression of a key enzyme of tryptophan metabolism,IDO.And this antidepressant-like effect may be mediated by 5-hydroxytryptamine and glutamate systems.

Extraction of Geniposide and Its Application in Anesthesiology

We tried to develop an efficient strategy to isolate geniposide from Gardenia fruit and use it to treat chronic pain. Eight extraction solvents and three extraction methods were respectively compared with each other for the efficiency of geniposide extraction from Gardenia fruit. Extraction with 80% ethanol under reflux was found to give the highest yield of geniposide. Orthogonal experiment was conducted to optimize the reflux conditions and the optimal conditions are a mass（of Gardeniajasminoides Ellis powder） to volume（of solvent, 80% ethanol） ratio of 1：7 and reflux twice for 60 rain each. The geniposide obtained by the reflux method was tested on the rats with chronic constriction injury（CCI） for pain relief. The results show that geniposide can enhance pain threshold and relieve pain via repressing tumor necrosis factor-a（TNF-a） expression in either ipsilateral or contralateral dorsal root gan- glion（DRG） cells. These results suggest that geniposide treatement may be an effective clinical method to relieve pain based on the mechanism of molecular biology at DRG due to nerve injury.

Comparative pharmacokinetics of baicalin and geniposide in juvenile and adult rats after oral administration of Qingkailing Granules

Objective:To explore the effect of age on Qingkailing Granules disposition by comparing the pharmacoknetics of geniposide and baicalin in juvenile and adult rats.Methods:A simple and rapid LC-MS/MS method was developed and validated to simultaneously determine geniposide and baicalin in rat plasma after a simple protein precipitation.The analytes were separated on an Agilent ZORBAX Extend-C18 column.The mobile phase consisted of acetonitrile and water with 0.1%(volume percent)formic acid at a flow rate of 0.6 mL/min.The ionization was conducted using an ESI source in negative ion mode.Multiple reaction monitoring was used for quantification at transitions of m/z 445.0→m/z 268.9 for baicalin,m/z 433.2→m/z 225.0 for geniposide,m/z 431.0→m/z341.0 for vitexin(IS).Juvenile and adult rats were administrated Qingkailing Granules(3 g/kg)orally.Plasma concentrations of baicalin and geniposide were determined by LC-MS/MS.Results:The linear ranges of the analytes were 1-1000 ng/mL for baicalin and 2-2000 ng/mL for geniposide.The method was successfully applied to compare the pharmacokinetics of the analytes between juvenile and adult rats after oral administration of Qingkailing Granules.AUC was bigger in adult rats,while t1/2 was longer in juvenile rats.Conclusion:These results suggested that the absorption and elimination of baicalin and geniposide in juvenile rats was lower than that in adult rats.Additional attention should be paid to the pharmacokinetic difference when Qingkailing Granules were used in children.

Effects of Bofutsushosan and Gardeniae Fructus on Diabetic Serum Parameters in Streptozotocin-Induced Diabetic Mice

Streptozotocin (STZ)-induced diabetic mice increased levels of serum glucose, triglyceride and cholesterol, and decreased level of serum insulin. Effects of Bofutsushosan (BOF: Pulvis ledebouriellae compositae: 防風通聖散) and its composed crude drug, gardeniae fructus (GF: 山梔子) were investigated on levels of these diabetic parameters (serum glucose, insulin, triglyceride and cholesterol) in STZ-diabetic mice. BOF and GF were extracted in 10 volumes of distilled water with an automatic extractor “Torobi”. STZ-induced diabetic mice with serum glucose level of over 600 mg/dl at 3 - 4 weeks after intravenous injection of 150 mg/kg STZ were used for experiments. BOF extract, GF extract, geniposide (a main constituent of GF), and glibenclamide were administered intraperitoneally into 3-hour-fasted STZ-diabetic mice. At 6 hours after administration, BOF extract (100 - 300 mg/kg) decreased high levels of serum glucose, triglyceride and cholesterol, and also increased low level of serum insulin in STZ-diabetic mice in a dose-dependent manner, respectively. Anti-diabetic drug glibenclamide (0.3 - 1 mg/kg) as positive control significantly decreased serum glucose and cholesterol levels, and increased serum insulin level in the diabetic mice. GF extract (30 - 300 mg/kg) decreased serum glucose, triglyceride and cholesterol levels but did not affect serum insulin level in the diabetic mice. Geniposide (10 - 100 mg/kg), decreased serum glucose level but did not affect serum insulin and triglyceride levels in the diabetic mice. These results demonstrated that intraperitoneally administrated BOF extract improved abnormal levels of serum glucose, insulin, triglyceride and cholesterol in the STZ-diabetic mice as being similar to glibenclamide. GF extract has an important role in a part of improving actions of BOF in the diabetic mice. The action of GF extract on serum glucose was parallel with the action of geniposide in the diabetic mice, supporting roles of geniposide in anti-hyperglycemic action of GF.

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.