Study on characteristic chromatogram of Chaenomeles sinensis and content determination of 3 flavones / 中国药房

OBJECTIVE To establish the characteristic chromatogram of Chaenomeles sinensis， determine the contents of rutin， hyperin and quercitrin， and to identify C. sinensis and C. speciosa. METHODS HPLC method was performed on Agilent 5 TC-C18 column， with acetonitrile-0.2% formic acid solution as the mobile phase for gradient elution， at the flow rate of 1.0 mL/min. The column temperature was 30 ℃ . The detection wavelength was 330 nm in characteristic chromatogram and 350 nm in content determination. The characteristic chromatogram of C. sinensis was established and similarity was evaluated by the Similarity Evaluation System for Chromatographic Fingerprint of TCM （2012 edition）. Hierarchical cluster analysis of 15 batches of C. sinensis （S1-S15） was performed by using SPSS 23.0 software. The contents of 3 flavones in 15 batches of C. sinensis and 7 batches of C. speciosa （S16-S22） were determined， while their characteristic chromatograms were compared. RESULTS The similarities of the characteristic chromatogram for 15 batches of C. sinensis ranged from 0.783 to 0.969， and 11 characteristic peaks were confirmed. Four constituents were identified as chlorogenic acid， rutin， hyperin and quercitrin. The medicinal materials in 15 batches of C. sinensis could be divided into 2 categories： S5-S8 were one category， and the others belonged to one category. The characteristic chromatogram of C. sinensis was obviously different from C. speciosa. The contents of rutin， hyperin and quercitrin in 15 batches of C. sinensis were 48.99-294.45， 3.49-102.55， 31.98-149.49 μg/g， respectively. The content of rutin in C. speciosa was lower than that in C. sinensis. None of hyperin （except for S20） and quercitrin were detected in C. speciosa. CONCLUSIONS The characteristic chromatogram and the method for content determination of 3 flavones in C. sinensis are established successfully and can be used for the quality control of C. sinensis and its identification from C. speciosa.

The osteoprotective effects and mechanisms of RA model mice by total saponins of Chaenomeles speciosa / 中国药房

OBJECTIVE To study the osteoprotective effects and possible mechanism of total saponins of Chaenomeles speciosa on rheumatoid arthritis （RA） model mice， and to provide reference for further development of anti-RA drugs. METHODS Seventy male DBA/1 mice were randomly divided into normal group， model group， low-dose and high-dose groups of C. speciose total saponins （60， 240 mg/kg）， Tripterygium wilfordii polyglycoside tablets group （positive control， 30 mg/kg）， with 14 mice in each group. In addition to the normal group， the other groups of mice were induced by glucose-6-phosphate isomerase mixed polypeptide to prepare RA model. The body weight， rear toes thickness and arthritis scores of each group were recorded； the synovial inflammation， bone and cartilage destruction of ankle joint tissues were observed by hematoxylin-eosin staining， tartrate- resistant acid phosphatase staining and safranin O-fast green staining； the contents of interleukin-6 （IL-6） in serum and tumor necrosis factor α （TNF-α）， IL-4 and IL-10 in ankle joint tissues were detected by ELISA； the expression levels of receptor activator of nuclear factor-κB ligand （RANKL）， receptor activator of nuclear factor-κB （RANK）， osteoprotegerin （OPG）， tumor necrosis factor receptor-associated protein 6 （TRAF6） and nuclear factor of activated T cells 1 （NFATC1） protein in ankle joint tissues were detected by Western blot assay. RESULTS At the end of administration， compared with normal group， the body mass of mice in the model group was significantly reduced （P＜0.05）， and the arthritis score and the thickness of the left and right rear toes were significantly increased （P＜0.05）； the ankle joint tissues of mice in the model group showed significant synovial proliferation and inflammatory infiltration， the number of osteoclasts increased significantly and significant destruction of cartilage tissue. The content of IL-6 in serum， the content of TNF-α， the protein expression levels of RANKL， RANK， TRAF6 and NFATC1 in the ankle joint tissues were increased significantly （P＜0.05）， while the contents of IL- 4 and IL-10， the protein expression level of OPG in the ankle joint tissues were decreased significantly （P＜0.05）. Compared with model group， above pathomorphological changes and the content/level of indicators of mice in each administration group were significantly improved （P＜0.05）. CONCLUSIONS Total saponins of C. speciosa may exert osteoprotective effects on RA model mice， the mechanism of which may be associated with reducing the contents of IL-6 and TNF-α， increasing the contents of IL-4 and IL-10， inhibiting the activation of RANKL/RANK/OPG signal pathway， thus inhibiting the proliferation of osteoclasts and promoting the repair of cartilage and bone tissue.

Protective effect of total triterpenoids from Chaenomeles speciosa against Helicobacter pylori-induced gastritis in mice / 中国中药杂志

This study investigated the protective effect of total triterpenoids from Chaenomeles speciosa against Helicobacter pylori(Hp)-induced gastritis in mice and explored its possible mechanism. The chronic atrophic gastritis(CAG) model mice were randomly divided into four groups of model, total triterpenoids from C. speciosa(50 and 100 mg·kg~(-1)) and triple therapy, with C57 BL/6 J mice without Hp infection taken as the normal group. Mice in the treatment groups were given corresponding drugs once a day for 4 weeks. Then the following indexes were detected: the contents of reactive oxygen species(ROS), monocyte chemotactic protein 1(MCP-1), keratinocyte chemokines(KC), TNF-α, IL-1β, IL-6, IL-18, IL-4 and IL-10 in blood and gastric tissue, the activities and contents of LDH, MPO, SOD, GSH-Px, CAT and MDA in gastric tissue and the activities of β-glucuronidase, β-galactosidase, cathepsins B and D in blood, gastric tissue and lysosome. Besides, the mRNA expression levels of Toll-like receptor 4(TLR4), myeloid differentiation factor 88(MyD88), Bcl-2, Bcl-xl, Bax and Bad in gastric tissue were determined by quantitative real-time PCR. Western blot was employed to detect the protein expression levels of TLR4, MyD88, p-IKKβ, p-IκBα, NOD-like receptor 3(NLRP3), apoptosis-associated speck-like protein(ASC), pro-caspase-1, caspase-1, thioredoxin-interacting protein(TXNIP), pro-IL-1β, pro-IL-18, Bcl-2, Bcl-xl, Bax, Bad, cytochrome C, apoptotic protease-activating factor-1(Apaf-1), pro-caspase-9, pro-caspase-3, cleaved-caspase-9, cleaved-caspase-3, poly(ADP-ribose) polymerase 1(PARP-1), cleaved-PARP-1 and cytosol and nucleus NF-κB p65 in gastric tissue. The results indicated that the total triterpenoids from C. speciosa significantly suppressed Hp proliferation, alleviated the damage to gastric mucosa and improved lymphocyte infiltration and gland atrophy. They were also effective in reducing the activities of β-glucuronidase, β-galactosidase, cathepsins B and D in blood and gastric tissue, elevating the activities of β-glucuronidase and cathepsin D in lysosomal organelles, decreasing the contents of ROS, MCP-1, KC, TNF-α, IL-1β, IL-6, IL-18 in blood, MDA content and MPO and LDH activities in gastric tissue and increasing the contents of IL-4 and IL-10 in blood and activities of SOD, CAT and GSH-Px in gastric tissue. Other phenomena were also observed after the treatment with total triterpenoids from C. speciosa, including the down-regulation of the mRNA and protein expression levels of TLR4, MyD88, Bax and Bad, the protein expression levels of p-IKKβ, p-IκBα, NLRP3, ASC, pro-caspase-1, caspase-1, TXNIP, pro-IL-1β, pro-IL-18, cytochrome C, Apaf-1, cleaved-caspase-9, cleaved-caspase-3, cleaved-PARP-1 and nuclear NF-κB p65, reduction of p-IKKβ/IKKβ and p-IκBα/IκBα ratios and up-regulation of the mRNA and protein expression levels of Bcl-2 and Bcl-xl, up-regulation of pro-caspase-9, pro-caspace-3, cytosol NF-κB p65 protein expression levels and Bcl-2/Bax and Bcl-xl/Bad ratios in gastric tissue. These aforementioned results suggest that the total triterpenoids from C. speciosa have significant protective effects against CAG induced by Hp, and its mechanism may be related to enhancing the function of endogenous antioxidant system, suppressing the oxidative stress and inflammatory reaction induced by Hp, correcting lysosomal dysfunction and inflammatory activation of TLR4/NF-κB/NLRP3 inflammasome signaling pathway and thus inhibiting mitochondria-mediated apoptosis.

Drying characteristics of Chaenomeles sinensis with different drying methods based on Dincer model / 中草药

Objective: Based on Dincer model, the drying characteristic of Chaenomeles sinensis under different drying condition was investigated in order to provide theoretical foundation for applying Dincer model to analyze heat and mass transfer during Chinese herbs drying process and select suitable drying technology and process. Methods: C. sinensis slice of thickness 12 mm was dried by the three different drying methods, namely air impingement drying, medium and short infrared waved drying and pulsed vacuum drying. Also, 9, 12 and 15 mm C. sinensis slices were dried under air impingement drying method. The drying characteristic, color value, rehydration ration, vitamin C (VC), general flavone, and microstructure were studied. Results: At the same drying temperature, the drying rate sorted in order of size was air impingement drying, medium and short infrared waved drying and pulsed vacuum drying and the drying activation energy was 43.10, 36.95 and 20.37 kJ/mol in corresponding. Decreasing slice thickness enhanced drying rate. The Weibull distribution model simulation result showed that the scale parameter α ranged from 47.85 to 324.51. Smaller α value meant short drying time. The shape parameter β was between 1.218 7 and 1.290 8 under air impingement drying as well as medium and short infrared waved drying method, which showed that drying was falling rate process controlled by internal moisture diffusion. However, the shape parameter β was between 1.218 7 and 1.290 8 under pulsed vacuum drying method, which illustrated that drying was controlled both by internal moisture diffusion and surface moisture evaporation. The calculated moisture diffusion coefficient was ranged from (1.66 × 10-8) to (1.13 × 10-7) m2/s and decreased as α increased. The Dincer model simulation result showed that the lag factor (G) was range from 1.135 6 to 1.337 6, which declared that there was a short raising rate drying period during the initial drying process. Heat transfer Biot number (Bi) value was between 1.171 4 and 136.041 2 and decreased as drying temperature increased. Effective moisture diffusion (Deff) value calculated by Diner model was range from (3.26 × 10-9) to (6.33 × 10-8) m2/s. At the same drying temperature, (Deff) value was larger than (D*), but smaller than (Dcal). Mass transfer (k) was ranged from (9.02 × 10-6) to (8.82 × 10-5) m/s and increased as drying temperature increased. Air impingement drying method was suitable for C. sinensis slice drying, and drying temperature of 60 ℃ and thickness of 12 mm was the most optimum drying process. Under above drying circumstance, the drying time, brightness L*, color difference value ΔE, VC, general flavone and rehydration ratio were 5 h, 62.80 ± 1.70, 19.62 ± 2.60, (1.107 8 ± 0.005 0) mg/g, (36.74 ± 0.60) mg/g and 7.11 ± 0.24, respectively. Conclusion: Such investigation result can provide theoretical foundation for applying Dincer model to describe heat and mass transfer characteristics during Chinese herbs drying and filtrating suitable C. sinensis slice drying method and process.

A new isobenzofuranone derivative from Chaenomeles sinensis and its antibacterial activity / 中国中药杂志

A new isobenzofuranone derivative was isolated from Chaenomeles sinensis by using various chromatographic techniques,including silica gel,Sephadex LH-20,MCI-gel resin and RP-HPLC. This compound was determined as 2,2-dimethyl-5-( 2-oxopropyl)-2 H-furo[3,4-h]chromen-7( 9 H)-one( 1) by NMR,MS,IR and UV spectra,and was also evaluated for its antibacterial activity. The results showed that it showed prominent antibacterial activity with MIC90 value of( 53. 7±4. 5) mg·L-1 for methicillin resistant Staphylococcus aureus( MRSA) strain. This value is close to that of levofloxacin [with MIC90 value( 50. 2± 4. 2) mg·L-1].

Therapeutic effect of total triterpenoids of Chaenomeles speciosa combined with omeprazole on gastric ulcer induced by indomethacin in rats / 中国中药杂志

The aim of this paper was to observe the combination therapy with total triterpenoids of Chaenomeles speciosa and omeprazole on indomethacin-induced gastric ulcer in rats, and explore its possible mechanism. Rats were randomly divided into normal group, model group, omeprazole monotherapy(3.6 mg·kg~(-1)) group, total triterpenoids of C. speciosa monotherapy(100 mg·kg~(-1)) group, total triterpenoids of C. speciosa and omeprazole combination therapy(100 mg·kg~(-1)+3.6 mg·kg~(-1)) group. Except for the normal group, the other groups were given indomethacin(20 mg·kg~(-1)) by oral once a day for 7 consecutive days. Then the treated groups were given corresponding drugs by gavage, once a day for 14 consecutive days. The next day after the last administration, half of the rats in each group were measured the gastric mucosal blood flow, gastric juice volume and serum TNF-α, IL-1β, IL-6, IL-4 and IL-10. After the remaining rats in each group were underwent pyloric ligation 4 hours after the last administration, the gastric endocrine volume, pH value and total acidity of gastric secretion were measured, then histological analysis was performed, MPO activity, cAMP content and histomorphological analysis were conducted. Real-time PCR was applied to detect the mRNA expressions of gastric tissue TNF-α,IL-1β, IL-6, IL-4, IL-10, VEGFA, A_(2A)R; the protein expressions of VEGFA, A_(2A)R, PKA, p-PKA, CREB, p-CREB, EGF, EGFR, p-EGFR, MUC6, TFF2 in gastric tissue were detected by Western blot. The results indicated that total triterpenoids of C. speciosa and omeprazole combination therapy might significantly increase gastric mucosal blood flow, gastric mucus volume, reduce gastric endocrine volume, secretion acidity and mucosal damage, decrease the levels of TNF-α,IL-1β and IL-6, increase the levels of IL-4 and IL-10 in blood and gastric tissue, inhibit the activity of MPO, increase the content of cAMP in gastric tissue, up-regulate the mRNA expressions of VEGFA, A_(2A)R and protein expressions of VEGFA, A_(2A)R, PKA, p-PKA, CREB, p-CREB, EGF, EGFR, p-EGFR, MUC6, TFF2 in gastric tissue, elevate p-PKA/PKA, p-CREB/CREB and p-EFGR/EFGR. Moreover, the combination therapy with total triterpenoids of C. speciosa and omeprazole was more obvious than those of two monotherapies. These aforementioned findings suggested that the combination therapy with total triterpenoids of C. speciosa and omeprazole on indomethacin-induced gastric ulcer have significant therapeutic effect on indomethacin induced gastric ulcer in rats, its mechanism might be related to regulating A_(2A)R/AKT/CREB, A_(2A)R/VEGFA, EGF/EGFR and MUC6/TFF2 signaling pathways, inhibiting pro-inflammatory factors, increasing gastric mucosal blood flow, up-regulating mucosal cell proliferation factors and promoting mucosal protective factors.

Multi-components determination and fringerprints analysis by HPLC in Chaenomeles thibetica / 中草药

Objective To establish a HSDE-HPLC-DAD-ESI-TOF/MS method for the rapid identification of chemical ingredients for Chaenomeles thibetica, and establish the multi-component quantitative fingerprint to evaluate its quality. Methods The separation was developed via an Agilent Zorbax SB-C18 column (250 mm × 4.6 mm, 5.0 μm) with a gradient elution at a temperature of 25 ℃, the acetonitrile was used as the mobile phase A and 0.5% glacial acetic acid and 10 mM ammonium acetate solution was used as the mobile phase B with a flow rate of 0.8 mL/min, and the detection wavelength was 280 nm. ESI-TOF/MS was used for the identification of chemical ingredients in C. thibetica. The fingerprint of C. thibetica was established based on the quantitative detection of multi-components. Results The HPLC fingerprint of C. thibetica has 30 common peaks, nine ingredients (chlorogenic acid, vanillic acid, caffeic acid, veratric acid, rutin, quercitrin, hyperin, oleanolic acid, and uosolic acid) of C. thibetica were identified, and the contents of them were determined. Similarity analysis shows that the similarity of 10 batches of C. thibetica is relatively close, while the similarity between C. thibetica and other varieties' is quite different. Conclusion HSDE-HPLC-DAD-ESI-TOF/MS method can be used for the rapid extraction and analysis of C. thibetica fresh samples. On this basis, the fingerprint analysis combined with similarity analysis can be used for quality evaluation of C. thibetica.

Analysis on moisture transfer laws of Chaenomeles sinensis during drying process based on low-field NMR / 中草药

Objective To study the moisture transfer laws of Chaenomeles sinensis in different drying processes. Methods Using the non-destructive and non-invasive technique of low field-nuclear magnetic resonance (LF-NMR), the transverse relaxation time (T2) inversion spectrum of C. sinensis slice was monitored under different drying methods (hot air drying, drying after evaporation, segmental drying and drying in the shade) to analyze the changes of moisture migration. Results There were three different types water that were detected in C. sinensis (free water > bound water > immobilized water). The internal water distribution and water content changed during drying process. The moisture changes were similar in hot air drying, drying after steaming, and drying in shade, the total water gradually decreased, and the combining degree between moisture and non-water components enhanced. Steaming promoted the water loss rate of C. sinensis slice, the water loss rate was higher in drying after steaming than in hot air drying, and the difference was significant (P < 0.05). During the intermittent drying, the conversion of different states of water would occur in order to return to a relatively stable equilibrium. During the low temperature drying process, immobilized water content decreased and free water content increased. The low-temperature drying has less damage to the tissue, which is more conducive to the conversion of immobilized water into free water, and thus the water dissipated faster. During the early of drying, high temperature caused tissue structure damage, the bonding force between water and non-aqueous tissue would be strengthened because of the tissue shrinkage. Conclusion The three different types water content and peak area in T2 was positively correlated. The LF-NMR technique would provide useful guides for the investigation of water distribution and variation of C. sinensis, which will provide a theoretical basis for C. sinensis processing.

Analysis on chemical constituents of fruit of Chaenomeles speciosa by UPLC-Q-Exactive Orbitrap-MS / 中草药

Objective An ultra-high performance liquid chromatography coupled with hybrid quadrupole-orbitrap mass spectrometry (UPLC-Q-Exactive Orbitrap-MS) method was developed to rapidly analyze and identify the chemical constituents from the fruit of Chaenomeles speciosa. Methods The analysis was performed on a Halo C18 column (100 mm × 2.1 mm, 2.7 μm) by gradient elution. The mobile phase consisted of 0.05% formic acid-acetonitrile and 0.05% formic acid-water at a flow rate of 0.3 mL/min. The column temperature was at 40 ℃. The information of the compounds was acquired in positive and negative mode. Results Totally 25 compounds were identified, including five kinds of free amino acids, 11 organic acids, two glycosides, three flavonoids, and four triterpenes. Conclusion The established method is accurate, rapid, and sensitive, which provides a reference for further clarifying the material basis of its efficacy and the selection of quality control indicators.

Chaenomelesalkaloid A, a new alkaloid from the fruits of Chaenomeles sinensis (Thouin) Koehne / 药学学报

The chemical constituents of the fruits of Chaenomeles sinensis (Thouin) Koehne were investigated using chromatographic methods, including Diaion HP-20, Toyopearl HW-40, MCI Gel CHP-20, ODS, Silica gel chromatography and semi-preparative-HPLC. Three compounds were isolated and their structures were elucidated with spectral data and physicochemical properties, which were identified as chaenomeles alkaloid A (1), ginsenine (2) and 1,2,3,4-tetrahydro-1-methyl-β-carboline-3-car-boxylic acid (3). Among those, compound 1 is a new alkaloid, compound 2 and 3 were isolated from this plant for the first time. To investigate the protective effect of compounds 1-3 on Rat adrenal pheochromocytoma (PC-12) injury induced by the β-amyloid protein (Aβ25-35). The results show that compounds 2 and 3 have a significant protective effect on the PC12 cells exposed to Aβ25-35.

Effect of total triterpenoids of Chaenomeles speciosa on PPARγ/SIRT1/NF-κBp65 signaling pathway and intestinal mucosal barrier of ulcerative colitis induced by DSS in mice / 中国中药杂志

To observe the effect of total triterpenoids of Chaenomeles speciosa on PPARγ/SIRT1/NF-κBp65 signaling pathway and intestinal mucosal barrier of ulcerative colitis induced by dextran sulfate sodium (DSS) in mice, C57BL/6 mice were randomly divided into normal group, model group, total triterpenoids of C. speciosa (50, 100 mg·kg⁻¹) groups and sulfasalazine (250 mg·kg⁻¹) group. The ulcerative colitis (UC) model was induced by orally administering 2.5% DSS to the experimental mice, and the corresponding drugs were given to each group 3 days before the administration with 2.5% DSS. The normal group and the model group were given the equal volume of 0.5% carboxymethyl cellulose sodium solution by gavage continuously for 10 days, q.d. The general conditions of the mice were observed on a daily basis, and the disease activity index (DAI) score was recorded. On the 10th day after the treatment, mice were put to death, the contents of TNF-α, IL-1β, IL-6, IFN-γ, IL-4 and IL-10 in the blood were detected, colon length was measured, colon mucosa damage index (CMDI) score was calculated, and MPO activity detection and histomorphology analysis were conducted. Real-time PCR was applied to detect the mRNA expressions of E-cadherin, occluding,MUC2 and TFF3; the protein expressions of SIRT1, IKKβ, p-IKKβ, IκBα, p-IκBα and cytosol and nucleus PPARγ, NF-κBp65 in intestinal tissue were detected by western blot. The results indicated that total triterpenoids of C. speciosa (50, 100 mg·kg⁻¹) could significantly improve the general conditions of UC mice, reduce the DAI, CMDI and histopathological scores, increase the colon length, reduce the colonic mucosa ulcers, erosion and inflammatory infiltration, restore the normal intestinal mucosal barrier function, reduce the contents of TNF-α, IL-1β, IL-6, IFN-γ, increase the contents of IL-4 and IL-10 in the blood, inhibit MPO activity in colon tissue, up-regulate the mRNA expressions of E-cadherin, occludin, MUC2 and TFF3 in colon tissue, down-regulate the protein expressions of cytosol PPARγ, tissue p-IKKβ, p-IκBα and nucleus NF-κBp65 in the colon tissue, decrease the p-IKKβ/IKKβ and p-IκBα/IκBα ratios, up-regulate the protein expressions of nucleus PPARγ, tissue SIRT1 and cytosol NF-κBp65 (<0.05 or <0.01, respectively), with a dose-effect relationship between the total triterpenoids of C. speciosa treated groups. These findings suggested that total triterpenoids of C. speciosa had a significantly therapeutic effect on UC mice induced by DSS, its mechanism might be related to the regulation of PPARγ/SIRT1/NF-κBp65 signaling pathway, the inhibition of pro-inflammatory factor formation and the up-regulation of protein expression of protective factors.

Rapid determination of oleanolic acid and ursolic acid in eight kinds of Chinese materia medica by HPLC / 中草药

Objective: To establish a method of rapid analysis for simultaneous determination of oleanolic acid and ursolic acid in Chinese materia medica (CMM) of Verbena officinalis, Ligustrum lucidum, Prunella vulgaris, Hedyotis diffusa, Patrinia scabiosaefolia, Eriobotrya japonica, Crataegus pinnatifida, Chaenomeles Fructus papaya. Methods: The analyses of preparation were conducted by accelerated solvent extraction (ASE), methanol was used as solvent extraction, and the static extraction time was 6 min. Separation was carried out on Acclaim C30 Thermo column with acetonitrile and 0.2% acetic acid (85:15) as mobile phase, flow rate was 0.3 mL/min, and UV detection wavelength was set at 205 nm. Results: After 10 min sample extraction time, oleanolic acid and ursolic acid reached baseline separation, and no sample matrix was interfered, the linear correlation coefficient was over 0.999, the average recovery between 95.8% and 102.7%; Compared to Chinese Pharmacopoeia 2015, the determination of average mass fraction difference was 3.7% and 5.8%. Conclusion: This method is simple, fast, and suitable for the analysis of these drugs, and it can be used for content determination of oleanolic acid and ursolic acid for eight kinds of CMM.

Effects of Extracted Active Components of Chaenomeles Speciosa on Non-alcoholic Fatty Liver Disease in Model Mice induced by High-fat–high-fructose Diet / 中国中医药信息杂志

Objective To study the effects of extracted active components of Chaenomeles Speciosa (EACCS) on non-alcoholic fatty liver disease (NAFLD) in mice; To discuss the possible molecular mechanism. Methods Forty male KM mice were randomized into four groups, namely normal group, model group, low-dose (50 mg/kg) EACCS group and high-dose (100 mg/kg) EACCS group. Except that the normal group was daily given routine diet, the other groups were given high-fat–high-fructose diet (HFFD). The mice were put to death 4 weeks later. Body weight, liver weight and serum TG were measured. HE and oil red O staining were used to observe liver tissue morphology. RT-PCR and Western blot were used to detect the expression of lipid metabolism related genes. Results Compared with the normal group, the liver size, liver index (P<0.01) and epididymal fat index (P<0.05) increased significantly;The ALT and GLU in serum increased (P<0.05), TG increased (P<0.05), and pathological findings showed significant steatosis; RT-PCR and Western blot showed that the expression levels of SIRT1 and FoxO1 mRNA decreased and the level of SERBP-1c increased in the model group. Compared with the model group, the hepatic lipid accumulation of EACCS groups was obviously improved, and the serum ALT, GLU, and TG levels significantly decreased, the expression levels of hepatic SIRT1 and FoxO1 mRNA increased. Conclusion EACCS has protective effects on NAFLD mice induced by HFFD, and its mechanism may be related to the activation of SIRT1-FoxO1 signaling pathway in the liver tissues.

Degranulation and anti-inflammatory effects of shikimic acid in Chaenomeles speciosa on peritoneal mast cells in rats / 中国免疫学杂志

Objective:To investigate the effect of Chaenomeles speciosa total extract shikimic acid(SA) on the degranulation of compound 48/80(C48/80) stimulated rat peritoneal mast cells and the anti-inflammatory.Methods: Qualitative analysis of shikimic acid in the total extract of Chaenomeles speciosa by high performance liquid phase;RPMC were identified by toluidine blue,immunohistochemistry and electron microscopy;CCK-8 method was used to detect the proliferation of RPMC in each concentration group.The release rate of β-hexosidase was determined by substrate method.The content of histamine in supernatant was detected by ELISA.Results: SA had no significant effect on the growth of RPMC at the concentration of 0-90 μg/ml.Compared with the positive control group,SA could effectively inhibit the release of β-hexosidase and inhibit the secretion of histamine.Conclusion: SA inhibits the release of histamine in inflammatory mediators by inhibiting the degranulation of RPMC stimulated by C48/80,and then exerts anti-inflammatory effects.

Study on DNA barcoding of medicinal plants in Chaenomeles Lindl / 中草药

Objective: To provide evidence for the molecular identification of medicinal plants in Chaenomeles Lindl. Methods: Nineteen ITS2 and psbA-trnH sequence samples of five species in Chaenomeles Lindl. were PCR amplified and sequenced. The intra-specific and inter-specific K-2P distances were calculated, and cluster analysis was performed using Neighbor-joining (NJ) method by MEGA 6.0. Results: The original plant of Chaenomeles Lindl. could be identified by the sequence differences of the ITS2 and psbA-trnH sequences. Different samples of Chaenomeles Lindl. were gathered together and could be distinguished by NJ tree. There were significant differences among the ITS2 secondary structures in five species of Chaenomeles Lindl. Conclusion: ITS2 and psbA-trnH are efficient barcodes for the authentication of plants of Chaenomeles Lindl.

Effects of Total Saponins of Chaenomeles speciosa on Degranulation Ratio of RBL-2H3 Mast Cells / 医药导报

Objective To study the effects of total saponins of Chaenomeles speciosa on release of β-hexosaminidase from rat basophilic leukemia-2H3 ( RBL-2H3 ) mast cells. Methods After rat RBL-2H3 mast cells were prepared, total saponins of Chaenomeles speciosa and the RBL-2H3 mast cells were co-cultured.The toxic effects of total saponins of Chaenomeles speciosa on mast cells were detected by MTT method, β-hexosaminidase release rate was measured by fluorescence quantitative spectrophotometric method, and cell supernatants of tumor necrosis factor-α( TNF-α) release were detected by ELISA. Results After total saponins of Chaenomeles speciosa were co-cultured with RBL-2H3 mast cells with different antigen stimulation, β-hexosaminidase release rates and the levels of TNF-α of mast cells significantly decreased compared with the control group. Conclusion Total saponins of Chaenomeles speciosa inhibit the degranulation of RBL-2H3 mast cells in a dose dependent manner, which provids basis for studying mechanism of inhibiting allergic reactions.

Content Determination of Chlorogenic Acid,Protocatechuic Acid and Total Phenolics in Chaenomeles sinersis from Different Areas / 中国药房

OBJECTIVE:To establish a method for the content determination of chlorogenic acid,protocatechuic acid and total phenolics in Chaenomeles sinensis,compare the content of total phenolics and 2 phenolic acids from different areas. METHODS:Wavelength switching HPLC method was conducted to determine the chlorogenic acid and protocatechuic acid. The column was Shim-pack CLC-ODS(M)with the mobile phase of acetonitrile-0.1% phoephoric acid(15∶85,V/V),the detection wavelength was 259 nm(for protocatechuic acid)and 325 nm(for chlorogenic acid)and the switching time was 14 min. With the index of proto-catechuic acid,Folin-Ciocalteau colorimetric method was conducted to determine the total phenolics. RESULTS:The 2 methods of quantitative analysis showed that the precision,repeatability,recoveries and standard curves were all validated by methodology. The mass fraction of total phenolics was 0.87%-3.77% with the average of 2.16%;the chlorogenic acid was 0.053%-0.387% with the average of 0.192% and the protocatechuic acid was 0.024%-0.541% with the average of 0.087%. The order of total phenolics content in C. sinensis from different areas was Yunnan>Anhui Xuancheng>Sichuan>Hubei and the order of total amount of chlo-rogenic acid and protocatechuic acid from different areas was the same as the total phenolics. There were differences among the con-tents of C. sinensis from different areas,however,the positive correlation was found between the content of total phenolics and the total amount of chlorogenic acid and protocatechuic acid with the pearson correlation coefficient of 0.719(P<0.01).CONCLU-SIONS:The established method is simple,accurate and reproducible and can be used for the content determination of chlorogenic acid,protocatechuic acid and total phenolics in chaenomelis fructus.

Research Progress in Chemical Constituents and Pharmacological Effects of Chaenomeles Speciosa Nakai and Chaenomeles / 中国药师

The studies on the chemical compositions and pharmacological actions of Chaenomeles speciosa Nakai were systemized and compared with those of the other plants of Chaenomeles in this paper. The pharmacological effects of the fruit of Chaenomeles inclu-ding anti-tumor, anti-oxidation, anti-inflammation and analgesia, antibacterial, hypoglycemic and hypolipidemic action and so on were reviewed to provide scientific basis for the further studies and utilization of Chaenomeles.

Effects of Different Processing Methods on the Contents of Oleanolic Acid and Ursolic Acid in Chaenomeles Medicinal Materials / 中国中医药信息杂志

Objective To investigate effects of different processing methods on the contents of oleanolic acid and ursolic acid in chaenomeles medicinal materials. Methods The contents of oleanolic acid and ursolic acid in samples processed by different processing methods in producing area were determined by HPLC with Agilent HC-C18 (2) chromatographic column (4.6 mm×250 mm, 5 μm), mobile phase of methanol-water-acetic acid-triethylamine (89∶11∶0.1∶0.05), detection wavelength of 210 nm, column temperature at 25 ℃, the flow rate of 1.0 mL/min. Results The total contents of oleanolic acid and ursolic acid in the samples prepared through 4 different processing methods were 0.86%, 0.93%, 1.29%, and 1.47%, respectively, showing that different processing methods in producing area had a significant effect on contents of oleanolic acid and ursolic acid in chaenomeles medicinal materials. Conclusion This study provides a basis for processing methods of chaenomeles medicinal materials in producing area.

Influence of Harvest Time on the Characters of Chaenomeles Spciosa (Sweet) Nakai / 中国药师

Objective:To study the influence of harvest time on the characters of Chaenomeles Spciosa ( Sweet) Nakai to determine the optimal harvest time with traditional characters of the Chinese medicine. Methods: In order to explore the optimal harvest time of Chaenomeles Spciosa ( Sweet) Nakai, the content of alcohol-soluble extract, weight and acidity of Chaenomeles Spciosa ( Sweet) Nakai with different harvest time were determined, the weather conditions in recent 3 years was summarized, and the drying process was also studied. Results:The average weight of Chaenomeles Spciosa(Sweet)Nakai was the lowest in June and highest in August, and the den-sity reached maximum in mid-July. During the whole harvest time, the content of alcohol-soluble extract was stable. The weather con-ditions from mid-July to late-July were with high temperature, dry and little rain, which was suitable for drying of Chaenomeles Spciosa ( Sweet) Nakai. Conclusion:The optimal harvest time of Chaenomeles Spciosa( Sweet) Nakai is confirmed in mid-July according to the traditional customs, drying conditions and characters of the Chinese medicine.