Study on quality standard of Gardenia jasminoides and its effective parts / 药学实践杂志

Objective To study the quality standard of Gardenia jasminoides and its effective parts. Methods TLC was used to identify Gardenia jasminoides and its effective parts. The heavy metals, harmful elements, and moisture in Gardenia jasminoides and its effective parts were examined. The content of Gardenia jasminoides and its effective parts was determined by high performance liquid chromatography. Results TLC method could be used to identify Gardenia jasminoides and its effective parts. The moisture content of Gardenia jasminoides and its effective parts were 8.4% and 3.2%, respectively. ICP-MS was used to determine the contents of five elements in Gardenia jasminoides and its effective parts simultaneously. There was a good linear relationship between arsenic, cadmium, copper, mercury, and lead in the range of 0~20, 0~10, 0~500, 0~5 and 0~20 ng/ml, respectively; The method detection limit of each metal element was 3.3×10−5~1.3×10−3 mg/kg. The relative standard deviation (RSD) of precision was 0.32%~0.82%. RSD values of each element content showed that the method had good repeatability. And the recoveries of arsenic, cadmium, copper, mercury, and lead were 103%~112%, 98%~99%, 98%~99%, 105%~106% and 100%~103%, respectively (n=3). The stability of each element was good within 8 h. The contents of the five elements were within the limits of the current edition of Chinese Pharmacopoeia. The standard curve equation of gardenia was Y=15860X+22543, r=0.9999, indicating that there was a good linear relationship of gardenia in the range of 20.16~322.6 μg/ml. The RSD of precision was 1.86%. RSD of the two samples were 2.38% and 2.60%, respectively, indicated that the method had good repeatability. The average recovery of Gardenia was 99.1% (n=6). The stability of the two solutions was good within 8 h. The contents of gardenia and its effective parts were 5.71% and 34.2%, respectively. Conclusion The research on the quality of Gardenia jasminoides effective parts was carried out based on the research on the quality of Gardenia jasminoides, and the results met the requirements. Therefore, the method established in this experiment could control the quality of Gardenia jasminoides and its effective parts simultaneously.

Systematic analysis of MYB transcription factors related to the geniposide biosynthesis in Gardenia jasminoides Ellis based on whole genome / 药学学报

MYB transcription factors are involved in the regulation of various secondary metabolites biosynthesis. Gardenia jasminoides Ellis is the commonly used Chinese herbal medicine, and its main active ingredient is geniposide. Here, leaves and flower buds at different developmental stages of G. jasminoides were used to explore MYB transcription factors related to geniposide biosynthesis based on genome and transcriptome analysis. Transcriptome data analysis showed that, different from the expression of the common pathway genes for terpenoid biosynthesis, the expression level of genes in the specific pathway of geniposide biosynthesis was significantly higher in flower buds than in leaves, which was the same as the organ accumulation pattern of this component. And the promoter regions of geraniol synthase, iridoid synthase and geniposidic acid methyltransferase involved in the specific pathway all contained multiple MYB-binding sites. A total of 105 MYB transcription factors were obtained by annotating the coding genes of G. jasminoides, which were divided into 68 1R-MYB, 33 R2R3-MYB, 3 3R-MYB and 1 atypical MYB transcription factor according to the number of conserved domain. Based on the analysis of phylogenetic tree and quantitative real-time PCR, three candidate MYB transcription factors related to geniposide biosynthesis were selected, including potential positive regulation factor GjMYB23 and negative regulation factors GjMYB31 and GjMYB73. The results of this study will lay a foundation for searching the regulation of geniposide biosynthesis and further analysis of the quality formation mechanism of G. jasminoides, so as to promote the breeding of excellent varieties of G. jasminoides.

Research progress on the mechanism of iridoids from Gardenia jasminoides against Alzheimer ’s disease / 中国药房

Alzheimer’s disease （AD）is a common latent neurodegenerative disease ，which is characterized by cognitive impairment，loss of learning and memory function ，abnormal behavior and dementia. At present ，there is no specific drug to effectively prevent or reverse AD. Gardenia jasminoides is the dried and mature fruit of G. jasminoides J. Ellis ，a gardenia plant in Rubiaceae. Its chemical components mainly include iridoids ，triterpenoids，organic acids and volatile oils ，among which iridoids are the main active components of G. jasminoides . This paper summarizes the researches on the mechanism of iridoids from G. jasminoides against AD at home and abroad in recent years ，in order to provide reference for the development of new drugs against AD.

Investigation of Neuroprotective Effect of Geniposide on Chronic Alcoholism Rats Based on Metabonomics / 中国实验方剂学杂志

ObjectiveMetabolomics was used to identify biomarkers of chronic alcoholism， and to evaluate the neuroprotective effect of geniposide， providing reference for the diagnosis and treatment of chronic alcoholism. MethodThe rat model of chronic alcoholism was established by intragastric administration of 50% ethanol with 8 mL·kg-1 for 14 days， and then increased to 12 mL·kg-1 for 21 days. Meanwhile， the intervention was performed by continuous gavage of geniposide （15 mg·kg-1） for 35 days. At the end of the experiment， the biochemical indexes and histopathological morphology of liver and brain tissues of rats were detected. Ultra performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry （UPLC-Q-TOF-MS） was used for urine metabonomics. The chromatographic conditions was as follows：ACQUITY UPLC™ HSS T3 column （2.1 mm×100 mm， 1.8 μm）， mobile phase of 0.1% formic acid acetonitrile solution （A）-0.1% formic acid aqueous solution （B） for gradient elution （0-2.5 min， 1%-11%A； 2.5-4.5 min， 11%-21%A； 4.5-7.0 min， 21%-40%A； 7.0-8.5 min， 40%-99%A； 8.5-10.5 min， 99%A； 10.5-10.6 min， 99%-1%A； 10.6-13.0 min， 1%A）， the flow rate of 0.4 mL·min-1. The conditions of mass spectrometry were electrospray ionization （ESI）， positive and negative ion modes， scanning range of m/z 50-1 200. Progenesis QI 2.0 and MassLynx 4.1 were used for data analysis， and biomarkers were identified by matching element composition and secondary fragments with Human Metabolome Database （HMDB）. ResultThe pathological results showed that on the 35th day of model replication， compared with the model group， the cortical neurons in the geniposide group showed a significantly improved state of disorder， nuclear pyknosis， hyperchromatism and cell membrane boundary blurred necrosis. The biochemical results showed that geniposide could significantly increase the activities of glutathione peroxidase （GSH-Px） and superoxide dismutase （SOD）， decrease the activity of acetylcholinesterase （AChE）， decrease the levels of β-endorphin （β-EP） and malondialdehyde （MDA）. A total of 48 biomarkers of chronic alcoholism were identified by metabonomics， involving seven metabolic pathways of tryptophan metabolism， phenylalanine metabolism， pentose and glucuronate interconversions， pyrimidine metabolism， ascorbate and aldarate metabolism， steroid hormone biosynthesis and purine metabolism. The main pathway is 5-hydroxytryptamine pathway of tryptophan metabolism. ConclusionBiomarkers related to nerve injury in chronic alcoholism are mainly derived from the 5-hydroxytryptamine metabolic pathway. Geniposide can regulate this pathway so as to improve oxidative stress in the brain and play a neuroprotective role.

PK2/PKR1 signaling pathway participates in geniposide protection against diabetic nephropathy in mice / 中国中药杂志

This study aimed to investigate the effects of geniposide(GP) on the expression of prokineticin(PK2) and prokineticin receptor 1(PKR1) in db/db mice with diabetic nephropathy(DN), so as to explore how the PK2 signaling pathway participated in the pathological changes of DN and whether GP exerted the therapeutic effect through this signaling pathway. Male mice were randomly divided into four groups, namely db/m, db/db, db/db+GP, and db/m+GP groups, with five in each group. The mice in the db/db+GP and db/m+GP groups were gavaged with 150 mg·kg~(-1) GP for eight successive weeks. Afterwards, all the mice were sacrificed and the renal tissues were embedded. The morphological changes in glomerulus and renal tubules were observed by Masson and PAS staining. The expression levels of PK2, PKR1, and Wilm's Tumor Protein 1(WT_1) in podocytes were detected by immunohistochemistry, and the protein expression levels of PK2 and PKR1 in mouse kidney by Western blot. The morphological results showed serious glomerular and tubular fibrosis(Masson), high glomerular and tubular injury score(PAS), increased glomerular mesangial matrix, thickened basement membrane, exfoliated brush border of renal tubules, decreased WT_1 in glomerular podocytes, and massive loss of podocytes in the db/db group. After administration with GP, the glomerular and tubular fibrosis was alleviated, accompanied by improved glomerular basement membrane and renal tubule brush edge, and up-regulated WT_1. As revealed by further protein detection, in the db/db group, the expression levels of PK2 and PKR1 and p-Akt/Akt ratio declined, whereas the ratio of Bax/Bcl-2 rose. Ho-wever, PKR2 and p-ERK/ERK ratio did not change significantly. After administration with GP, the PK2 and PKR1 expression was elevated, and p-Akt/Akt ratio was increased. There was no obvious change in PKR2, Bax/Bcl-2 ratio, or p-ERK/ERK ratio. All these have demonstrated that GP improves the renal damage in DN mice, and PK2/PKR1 signaling pathway may be involved in such protection, which has provided reference for clinical treatment of DN with GP.

Method validation of total iridoids in “Darmon” tablets by spectrophotometric method / Монголын эм зүй, эм судлал

Introduction@#A joint research team of the Drug Research Institute аndMonos pharm Co.ltd is conducting an experiment to produce of “Darmon” tablets.Idridoids are one of the predominant biological active compound in “Darmon” tablets and will be an important indicator of the quality of the drug.@*Objectives@#This is the first report on the determination of iridoids by spectrophotometric method in “Darmon” tablets.@*Methods@#The amount of total iridoids of “Darmon” tablets was confirmed by spectrophotometry and the absorbance was measured at 238 nm. Geniposide (98%, Xilong Scientific Co., Ltd) was used as the standard substance.@*Results@#The developed spectrophotometric method showed good linearity (R²=0.9989), high precision (RSD<2%) and a good recovery (96.01-104.48%). All the validation parameters of the spectrophotometric method were found to be within the permissible limits according to the ICH guidelines. @*Conclusions@#The method was robust, accurate and reliable for the quality control of “Darmon” tablets.

Combination of Geniposide and Eleutheroside B Exerts Antidepressant-like Effect on Lipopolysaccharide-Induced Depression Mice Model / 中国结合医学杂志

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-1 β (IL-1 β) 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-1 β content, inhibited the expression levels of NF- κ B 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-α content 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-1 ß content 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-1 β, 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.

Molecular mechanism of geniposide in regulating GLUT2 glycosylation in pancreatic β cells / 中国中药杂志

Type 2 diabetes mellitus( T2 DM) is a common chronic metabolic disease characterized by persistent hyperglycemia and insulin resistance. In pancreatic β-cells,glucose-stimulated insulin secretion( GSIS) plays a pivotal role in maintaining the balance of blood glucose level. Previous studies have shown that geniposide,one of the active components of Gardenia jasminoides,could quickly regulate the absorption and metabolism of glucose,and affect glucose-stimulated insulin secretion in pancreatic β cells,but the specific mechanism needs to be further explored. Emerging evidence indicated that glycosylation of glucose transporter( GLUT) has played a key role in sensing cell microenvironmental changes and regulating glucose homeostasis in eucaryotic cells. In this study,we studied the effects of geniposide on the key molecules of GLUT2 glycosylation in pancreatic β cells. The results showed that geniposide could significantly up-regulate the mRNA and protein levels of Glc NAc T-Ⅳa glycosyltransferase( Gn T-Ⅳa) and galectin-9 but had no signi-ficant effect on the expression of clathrin,and geniposide could distinctively regulate the protein level of Gn T-Ⅳa in a short time( 1 h) under the conditions of low and medium glucose concentrations,but had no significant effect on the protein level of galectin-9. In addition,geniposide could also remarkably affect the protein level of glycosylated GLUT2 in a short-time treatment. The above results suggested that geniposide could quickly regulate the protein level of Gn T-Ⅳa,a key molecule of protein glycosylation in INS-1 rat pancreatic βcells and affect the glycosylation of GLUT2. These findings suggested that the regulation of geniposide on glucose absorption,metabolism and glucose-stimulated insulin secretion might be associated with its efficacy in regulating GLUT2 glycosylation and affecting its distribution on the cell membrane and cytoplasm in pancreatic β cells.

Effect of Geniposide on Liver Injury and Bile Acid Profile in Serum, Liver and Excrement of Rats / 中国实验方剂学杂志

Objective:To investigate the hepatotoxicity of different doses of geniposide on the liver of rats and the effects on bile acid profile in serum， liver tissue and feces. Method:The 60 Sprague Dawley rats， half male and half female， were randomly divided into 5 groups according to body weight： blank group and four different doses （50， 100， 200， 400 mg·kg^-1） geniposide groups， 12 rats in each group. The rats were treated by gavage once a day for 7 consecutive days， and the serum， liver and cecal contents were collected on the 8^th day of treatment. The activities of alanine aminotransferase （ALT）， aspartate aminotransferase （AST） and alkaline phosphatase （ALP）， the contents of albumin （ALB）， total bilirubin （TBIL）， total bile acid （TBA）， creatinine （Crea） and carbamide （Urea） were detected in each group. The sections of liver tissue were stained with hematoxylin-eosin（HE）， and the protein expressions of cytokeratin 7（CK7） and cytokeratin 19（CK19） were detected by immunohistochemistry. The protein expressions of CK7 and CK19 in the liver tissue were detected by Western blot. And the mRNA expressions of cholesterol 7<italic>α</italic>-hydroxylase （CYP7A1）， cholesterol 27<italic>α</italic>-hydroxylase （ CYP27A1） and cholesterol 12<italic>α</italic>-hydroxylase （CYP8B1） were detected by real-time PCR. The contents of 18 kinds of bile acids in serum， liver and cecal contents were determined by ultra-performance liquid chromatography-mass spectrometry（UPLC-MS）. Result:Compared with the control group， TBIL level in each dose of geniposide group was increasesd significantly （<italic>P</italic><0.01）. ALT， AST activity and TBA content in 400 mg·kg^-1 geniposide group were increased significantly （<italic>P</italic><0.05， <italic>P</italic><0.01）. HE staining showed that， compared with control group， there was bile duct reaction in the portal area and inflammatory cells infiltrate around bile duct in 200 mg·kg^-1 and 400 mg·kg^-1 geniposide groups， especially 400 mg·kg^-1. The expressions of CK7 and CK19 in liver tissue of 400 mg·kg^-1 geniposide group were significantly higher than those in the control group （<italic>P</italic><0.05， <italic>P</italic><0.01）. Compared with the control group， the contents of glycoursodeoxycholic acid （GUDCA） and glycohyodeoxycholic acid （GHDCA） in liver tissue of 400 mg·kg^-1 geniposide group decreased significantly （<italic>P</italic><0.05， <italic>P</italic><0.01）， the contents of sodium taurochenodeoxycholate （TCDCA）， hyodeoxycholic acid （HDCA）， cholic acid （CA） and chenodeoxycholic acid （CDCA） in liver tissue increased significantly （<italic>P</italic><0.01）， the contents of glycocholic acid hydrate （GCA）， glycochenodeoxycholic acid （GCDCA）， glycodeoxycholic acid hydrate （GDCA）， glycocholic acid （GLCA）， tauroursodeoxycholic acid （TUDCA）， GUDCA， GHDCA， ursodeoxycholic （UDCA） and taurolithocholic acid （TLCA） decreased， the proportions of TCDCA， HDCA， CA， CDCA and deoxycholic acid （DCA） in liver tissue increased， the contents of GHDCA and lithocholic acid （LCA） in serum decreased significantly （<italic>P</italic><0.01）， while sodium taurohyodeoxycholate hydrate （THDCA）， taurocholic acid （TCA）， GCA， TCDCA， UDCA， CA， CDCA， DCA in serum decreased significantly （<italic>P</italic><0.05， <italic>P</italic><0.01）. The contents of CA， UDCA， CA， CDCA and DCA increased significantly （<italic>P</italic><0.05）， the ratio of CA/DCA increased significantly （<italic>P</italic><0.05）， and the ratio of CA and CDCA increased by 19.60% and 4.63%， respectively； Compared with the control group， the contents of all bile acids in cecal contents of 400 mg·kg^-1 were decreased， and the contents of GCA， UDCA， HDCA， GCDCA， GDCA， TLCA， GLCA， CDCA， DCA and LCA were decreased significantly （<italic>P</italic><0.05， <italic>P</italic><0.01）. In addition， real-time PCR results showed that the mRNA expressions of CYP7A1， CYP27A1 in the 400 mg·kg^-1 geniposide group were significantly higher than those in the control group （<italic>P</italic><0.05， <italic>P</italic><0.01）. Conclusion:The 400 mg·kg^-1geniposide can cause obvious hepatotoxicity in rats， and the bile acid profile in liver， serum and excrement changes significantly， and the changes of the each bile acid in liver， serum and feces are different. However， the causal relationship between the gardenoside-induced liver injury and the changes in bile acid profile are<italic> </italic>not clear. It needs to be further studied.

Optimization of the Quality Control Method of Qingkailing Dropping Pills / 中国药学杂志

OBJECTIVE: To optimize and improve the quality standard of Qingkailing dropping pills. METHODS Lonicerae Japonicae Flos, geniposide in Gardeniae Fructus and baicalin in Scutellariae Radix of Qingkailing dropping pills were identified by TLC and HPLC respectively. Cholic acid and hydrodeoxycholic acid were determined by HPLC equipped with an ELSD and a column (4.6 mm×250 mm, 5 μm) packed with ODS bonded silica gel (5 μm particle size). The mobile phase was a mixture of methanol, acetonitrile and 0.1% formic acid (68∶17∶15, V/V/V) and the flow rate was 1.0 mL•min-1. Geniposide in Gardeniae Fructus of Qingkailing dropping pills was determined by HPLC equipped with a DAD (238 nm) and a column (4.6 mm×250 mm) packed with ODS bonded silica gel (5 μm particle size). The mobile phase was a mixture of acetonitrile and 0.05% phosphoric acid (10∶90, V/V) and the flow rate was 1.0 mL•min-1. Baicalin in Scutellariae Radix of Qingkailing dropping pills was determined by HPLC equipped with a DAD (275 nm) and a column (4.6 mm×150 mm) packed with ODS bonded silica gel (5 μm particle size), the mobile phase was a mixture of methnol, water and phosphoric acid (47∶53∶0.2, V/V/V), and the flow rate was 1.0 mL•min-1. RESULTS: The developed TLC spots were clear with good reproducibility. Hydrodeoxycholic acid, cholic acid, geniposide and baicalin showed good linear relationship in the ranges of 0.505 0-6.312 μg, 0.515-6.440 μg, 0.029 12-0.582 4 μg, and 0.109 2-1.092 μg, respectively. The average recoveries (n=6) were 101.5%, 101.6%, 98.61% and 99.77%, with RSDs of 1.7%, 1.5%, 1.42% and 0.79%, respectively. CONCLUSION: The method is simple, rapid, accurate, with low toxicity, and can be used to control the quality of Qingkailing dropping pills more effectively and comprehensively.

Metabolomic study of Gardenia jasminoside var. radicans and geniposide on degranulation model of RBL-2H3 cells / 中草药

Objective: To study the intervention effect of Gardenia jasminoside var. radicans and its main effective component of geniposide on the degranulation model of RBL-2H3 cells based on metabolomics. Methods: The changed metabolite profile of RBL-2H3 cells was detected by UPLC-QTOF-MS; PCA (principal component analysis) and OPLS-DA (orthogonal partial least squares discriminant analysis) in SIMCA software were used to select the potential biomarkers. Meanwhile, the clustering and heat map analysis for those potential biomarker levels were carried out by MEV software. Result: A total of 54 and 46 relevant biomarkers of G. jasminoside var. radicans and geniposide were selected, of which 31 biomarkers enriched in five disturbed metabolite pathways, including glycine, aspartic acid and glutamate metabolism, glutathione metabolism, histamine metabolism, energy metabolism, and nicotinamide metabolism pathways. Conclusion: G. jasminoside var. radicans and geniposide exerts the inhibitory effect on the degranulation model of RBL-2H3 cells by regulating histamine metabolism, oxidative stress and energy metabolism, and geniposide was one of the main efficacious substance basis of G. jasminoside var. radicans.

Simultaneous determination of six components in Mongolia medicine Gurigumu-13 Pills by QAMS / 中草药

Objective: To establish a quantitative analysis of multi-components by single marker (QAMS) method for the simultaneous determination of six components of gallic acid, hydroxysafflor yellow A, geniposide, ellagic acid, costunolide and dehydrocostus lactone in Gurigumu-13 Pill, which is proved to be a scientific and feasible method in the quality analysis in Gurigumu-13 Pill. Methods: The relative factor (fs/i) of gallic acid, ellagic acid, hydroxysafflor yellow A, costunolide and dehydrocostus lactone were established by HPLC method with geniposide as internal standard, which were used to calculate the content of five constituents in the samples of Gurigumu-13 Pill. Meanwhile, external standard method (ESM) was used to calculate the content of six constituents. The difference between QAMS and ESM was analyzed to evaluate the accuracy of QAMS. Results: The fs/i of gallic acid, hydroxysafflor yellow A, ellagic acid, costunolide and dehydrocostus lactone were 0.481 0, 0.906 4, 0.170 9, 0.971 2 and 1.261 5, respectively. The content determination results of six batches of Gurigumu-13 Pill were calculated by the method of QAMS and ESM, with no significant difference in RSD < 2.0%. Conclusion: The fs/i established in the QAMS method with geniposide as the internal reference substance is accurate and simple. The QAMS method can be used for the multi-index quality evaluation of Gurigumu-13 Pill.

Comparative study on differences of Gardeniae Fructus and its stir-baked prepared slices based on whole and partial fingerprint and multivariate statistical analysis / 中草药

Objective: To compare the differences of Gardeniae Fructus and its stir-baked preparation and screen out the differential markers, and provide reference for the establishment of quality standard. Methods: The whole and partial fingerprints of Gardeniae Fructus and its stir-baked prepared slices were established by HPLC-UV analysis. And the fingerprints were statistically analyzed using hierarchical clustering analysis (HCA), principle component analysis (PCA), and orthogonal partial least squares-discriminant analysis (OPLS-DA), then the characteristic markers were screened out. And then content of 6-α-hydroxygeniposide, gentioside, geniposide, crocin I, and crocin II was determined. Results: A total of 15 peaks were acquired for Gardeniae Fructus and stir-baked Gardeniae Fructus respectively, 12 peaks of which were the common peaks. The similarity of whole and partial reference fingerprints of Gardeniae Fructus and its stir-baked prepared slices was 0.991 and 0.880, respectively, and partial fingerprint can successfully distinguished Gardeniae Fructus from its stir-baked prepared slices. HCA and OPLS-DA results showed that significant differences were observed between Gardeniae Fructus and stir-baked Gardeniae Fructus with peak 3, peak 5, peak 9 (geniposide), and peak 11 (crocin I), which being identified as the main differential markers. Contents of geniposide, crocin I and crocin II in Gardeniae Fructus were higher than those in stir-baked Gardeniae Fructus, while 6-α-hydroxygeniposide was lower. Conclusion: The composition was significantly different before and after stir-baked. The established partial fingerprints combined with multi-component determination method can effectively distinguish them. And the differential markers obtained by multivariate statistical analysis can provide reference for the selection of quality markers.

Spectrum-effect Relationship of Choleretic Effect of Gardeniae Fructus Extract Based on Principal Component Analysis and Gray Correlation Analysis / 中国实验方剂学杂志

Objective:To investigate the effective substance of the choleretic effect of Gardeniae Fructus，and analyze the relationships between its choleretic effect and the HPLC fingerprint chromatogram. Method:HPLC method was applied to establish the fingerprint chromatography of 8 batches of Gardeniae Fructus at different harvest periods. The flow，the content of bile acid，bilirubin and cholesterol in bile were tested，and then the principal component analysis was used to comprehensively evaluate the total choleretic effects of Gardeniae Fructus. After the relationships between the relative peak area of the common peaks and the choleretic effects were explored using grey relationship analysis method，the spectrum-effect relationship of Gardeniae Fructus was established. Result:The order of the contribution of the chemical components to the choleretic effect at the common peaks was as follow（r>0.8）：P9>P14>P26>P4>P30>P6>P1>P10>P5>P24. Among all peaks，the full wavelength scanning results implied that the peaks 9，14 and 4 might be iridoids, and the peaks 26，30 and 24 might be crocins. By comparing with the standard substances，the peak 9 was finally identified as geniposide. Conclusion:The choleretic effect of Gardeniae Fructus may be the results of multiple components and pathways，and the main components in Gardeniae Fructus with the choleretic effect was from geniposide. In conclusion，these results provide a reference for investigating the material basis of choleretic effect of Gardeniae Fructus.

Comparative pharmacokinetics of baicalin and geniposide in juvenile and adult rats after oral administration of Qingkailing Granules / 中草药·英文版

Objective: To explore the effect of age on Qingkailing Granules disposition by comparing the pharmacokinetics 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/z 341.0 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 Geniposide on the Levels of Blood Sugar and Lipids in Streptozocin-Induced Diabetic Rats / 中国药学杂志

OBJECTIVE: To investigate the regulation of geniposide on blood sugar and lipids in diabetic rats. METHODS: ELISA assay was used to determine the content of insulin in vitro and in vivo, and MTT was used to determine the cell viability.RESULTS Geniposide prevented pancreatic beta damage induced by high glucose, and attenuated glucose-stimulated insulin secretion in the presence of 25 mmol•L-1 glucose. Furthermore, geniposide ameliorated insulin resistance, and decreased the contents of total cholesterol (TC) and triglyceride (TC) in STZ-induced diabetic rats. CONCLUSION: Geniposide is beneficial to improve the dysfunction of pancreatic beta cells induced by high glucose, and might be a promising lead compound for the treatment of diabetes.

Research progress on anti-inflammatory activity of three kinds of iridoid glycosides in Chinese materia medica / 中草药

Iridoid glycosides are widely distributed in Chinese materia medica (CMM) with various biological activities such as anti-inflammation. They are also used as quality control constituents in some Chinese medicines. Iridoid glycosides are usually divided into nine-carbon skeleton iridoid glycoside type, ten-carbon skeleton iridoid glycoside type, and secoiridoid glycoside type. In this paper, 15 representative iridoid glycosides from three types which are received extensive attention (including geniposide, catalpol, gentiopicroside, etc) have been selected. Their anti-inflammatory effects and possible related mechanisms are summarized to find out the acting feature of different types. Through comparing the structures and function characteristics, it was concluded that the anti-inflammatory effects of iridoid glycosides were mostly related to NF-κB pathway and MAPK pathway. They have obvious inhibitory effects on TNF-α, IL-6, and IL-1β inflammatory factors, some of which could play a role by reducing the expression of iNOS and COX-2 in NLRP3, Nrf2/HO-1, PI3K, and other pathways. From the structure-activity relationship, the double bond on the cyclopentane, the C-11 substituent and the bond formation after ring opening in iridoid glycosides all have important effects on its anti-inflammatory activities.

Optimization of extraction technology of Guizhi Zhumian Capsule by orthogonal design and pharmacodynamics test / 中草药

Objective: To optimize the extraction technology of Guizhi Zhumian Capsules (GZC). Methods: Based on the analysis of the single factor experiment Results:, the orthogonal test method was adopted to study the three factors including the amount of water added, extraction time, and extraction frequency by taking the dry extract yield of the medicinal materials and transfer rate of geniposide as indicators to optimize the water extraction process of GZC. To optimize the alcohol precipitation process of GSC, these factors including the concentration of the medicinal materials, alcohol content, and time of alcohol precipitation were investigated. The extracts before and after alcohol precipitation were compared by improving sleep pharmacodynamics. Results: The best water extraction technology of GZC was decocted three times with 10 times of water, 0.5 h each time. The optimal alcohol precipitation process was to concentrate the filtrate of water extraction to 1 mL, which was equivalent to 1 g of the original medicinal materials, with 80% alcohol content and 12 h alcohol precipitation time. Compared with the negative control group, the water extract group of GZC could prolong the sleep time of pentobarbital sodium mice and increase the sleep rate of mice under the lower dose of pentobarbital sodium valve (P 0.05). Conclusion: The pharmaceutical efficacy of the extract before and after alcohol precipitation was different. In order not to affect the drug effect, the water extraction process was finally selected as the best extraction process.

Fingerprinting study on Niuhuang Shangqing Pills based on comprehensive two-dimensional liquid chromatography / 中草药

Objective: To develop a new three-dimensional fingerprinting method and its assessing Methods: based on comprehensive two-dimensional liquid chromatography using Niuhuang Shangqing Pills (NSP) as an example. The developed method could offer new method for the quality control of NSP. Methods: In first dimension, the separation was achieved with an Acquity UPLC HSS CYANO column (100 mm × 2.1 mm, 1.8 μm), methanol-0.1% formic acid in water were used as mobile phases, flow rates were 0.1 mL/min. In second dimension, the separation was achieved with a Kinetex Phenyl-Hexyl column (50 mm × 3 mm, 2.6 μm), acetonitrile-0.1% formic acid in water were used as mobile phases, flow rates were 1.5 mL/min, detection wavelength was set at 254 nm, and acquiring frequency was at 12.5 Hz. Column temperature for each dimension was 40 ℃ and volume of loop linking the two dimensions was 100 μL. Three similarity-calculating Methods:, Euclidean Distance, Cosine, and Correlation Coefficient, were employed to assess the similarities among the 21 samples on the market using medians with arithmetic means of peak volumes of the common peaks as control fingerprints. Results: The three-dimensional fingerprints of 21 batches of NSP samples on the market were developed; Eighteen common peaks were assigned and five of them were identified, which were geniposide (1), pulegone (8), baicalin (9), imperatorin (15), and wogonin (16). Conclusion: A three-dimensional fingerprinting method and its assessing Methods: based on comprehensive two-dimensional liquid chromatography using NSP as an example were successfully developed for the first time, suggesting that it is a feasible method for developing fingerprints for Chinese materia medica. This work improves and supplements the traditional liquid-chromatography fingerprints.

Quality evaluation of Gardeniae Fructus from different areas by using chemical pattern recognition based on combination of chromatographic fingerprints and multi-component content / 中草药

Objective: To evaluate the quality of Gardeniae Fructus from different producing areas and optimize the best place of production by using chemical pattern recognition method based on HPLC fingerprint and multi-component content determination. Methods: The RP-HPLC using acetonitrile-0.1% phosphoric acid aqueous solution as mobile phase based on the wavelength switching (234 nm and 440 nm) and gradient elution method was developed to evaluate the quality of 30 batches of Gardeniae Fructus from 10 different producing areas. The combination of chromatographic fingerprints and quality determination of five active ingredients, as well as chemometrics including hierarchical cluster analysis (HCA) and principal component analysis (PCA), were further employed for the quality assessment. Results: The fingerprint similarity of 30 batches of Gardeniae Fructus from 10 different producing areas were all above 0.95. The content ranges of deacetylated methyl oxalate, genipin gentiobioside, geniposide, crocin I, and crocin II were 0.61-4.26 mg/g, 1.73-12.92 mg/g, 51.79-82.76 mg/g, 5.03-12.80 mg/g, and 0.71-2.28 mg/g. Samples can be divided into three categories by HCA: The first category is Yongzhou in Hunan, Yong’ an in Hunan, Fengcheng in Jiangxi, Hukou in Jiangxi, Guangde in Anhui, and Fuding in Fujian; The second category is Lichuan in Jiangxi, Jurong in Jiangsu and Tanghe in Henan; And the third category is Lu’ an in Anhui. PCA was used to further evaluate the quality differences of the samples from different producing areas, and it was found that the medicinal materials from Lu’ an in Anhui, Hukou in Jiangxi, and Yong’ an in Hunan were of superior quality. Conclusion: The quality of Gardeniae Fructus from different producing areas was different to a certain extent, while the quality of same batches was stable. By combination of fingerprint and content determination, the chemical pattern recognition method can be used to evaluate the quality of the Gardeniae Fructus comprehensively. The establishment of this method provided an effective reference for the quality control and evaluation of Gardeniae Fructus.