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.

Effects and mechanisms of total flavones of Abelmoschus manihot in attenuating diabetic tubulopathy by targeting endoplasmic reticulum stress-induced cell apoptosis / 中国中药杂志

Renal tubular injury in patients with diabetic kidney disease(DKD) may be accompanied by glomerular and microvascular diseases. It plays a critical role in the progression of renal damage in DKD, and is now known as diabetic tubulopathy(DT). To explore the multi-targeted therapeutic effects and pharmacological mechanisms in vivo of total flavones of Abelmoschus manihot(TFA), an extract from traditional Chinese medicine for treating kidney disease, in attenuating DT, the authors randomly divided all rats into four groups: a normal control group(normal group), a DT model group(model group), a DT model+TFA-treated group(TFA group) and a DT model+rosiglitazone(ROS)-treated group(ROS group). The DT rat model was established based on the DKD rat model by means of integrated measures. After successful modeling, the rats in the four groups were continuously given double-distilled water, TFA suspension, and ROS suspension, respectively by gavage every day. After 6 weeks of treatment, all rats were sacrificed, and the samples of their urine, blood, and kidneys were collected. The effects of TFA and ROS on various indicators related to urine and blood biochemistry, renal tubular injury, renal tubular epithelial cell apoptosis and endoplasmic reticulum stress(ERS), as well as the activation of the protein kinase R-like endoplasmic reticulum kinase(PERK)-eukaryotic translation initiation factor 2α(eIF2α)-activating transcription factor 4(ATF4)-C/EBP homologous protein(CHOP) signaling pathway in the kidney of the DT model rats were investigated. The results indicated that hypertrophy of renal tubular epithelial cells, renal tubular hyperplasia and occlusion, as well as interstitial extracellular matrix and collagen deposition occurred in the DT model rats. Moreover, significant changes were found in the expression degree and the protein expression level of renal tubular injury markers. In addition, there was an abnormal increase in tubular urine proteins. After TFA or ROS treatment, urine protein, the characteristics of renal tubular injury, renal tubular epithelial cell apoptosis and ERS, as well as the activation of the PERK-eIF2α-ATF4-CHOP signaling pathway in the kidney of the DT model rats were improved to varying degrees. Therein, TFA was superior to ROS in affecting the pathological changes in renal tubule/interstitium. In short, with the DT model rats, this study demonstrated that TFA could attenuate DT by multiple targets through inhibiting renal tubular ERS-induced cell apoptosis in vivo, and its effect and mechanism were related to suppressing the activation of the PERK-eIF2α-ATF4-CHOP signaling pathway in the kidney. These findings provided preliminary pharmacological evidence for the application of TFA in the clinical treatment of DT.

Effects and mechanisms of total flavones of Abelmoschus manihot in improving insulin resistance and podocyte epithelial-mesenchymal transition in diabetic kidney disease based on IRS1/PI3K/Akt pathway / 中国中药杂志

This study aimed to explore the effects and mechanisms of total flavones of Abelmoschus manihot(TFA), the extracts from traditional Chinese medicine indicated for kidney diseases, on insulin resistance(IR) and podocyte epithelial-mesenchymal transition(EMT) in diabetic kidney disease(DKD), and further to reveal the scientific connotation. Thirty-two rats were randomly divided into a normal group, a model group, a TFA group, and a rosiglitazone(ROS) group. The modified DKD model was induced in rats by methods including high-fat diet feeding, unilateral nephrectomy, and streptozotocin(STZ) intraperitoneal injection. After modeling, the rats in the four groups were given double-distilled water, TFA suspension, and ROS suspension correspondingly by gavage every day. At the end of the 8th week of drug administration, all rats were sacrificed, and the samples of urine, blood, and kidney tissues were collected. The parameters and indicators related to IR and podocyte EMT in the DKD model rats were examined and observed, including the general condition, body weight(BW) and kidney weight(KW), the biochemical parameters and IR indicators, the protein expression levels of the key signaling molecules and structural molecules of slit diaphragm in the renal insulin receptor substrate(IRS) 1/phosphatidylinositol 3-kinase(PI3K)/serine-threonine kinase(Akt) pathway, foot process form and glomerular basement membrane(GBM) thickness, the expression of the marked molecules and structural molecules of slit diaphragm in podocyte EMT, and glomerular histomorphological characteristics. The results showed that for the DKD model rats, both TFA and ROS could improve the general condition, some biochemical parameters, renal appearance, and KW. The ameliorative effects of TFA and ROS were equivalent on BW, urinary albumin(UAlb)/urinary creatinine(UCr), serum creatinine(Scr), triglyceride(TG), and KW. Secondly, they could both improve IR indicators, and ROS was superior to TFA in improving fast insulin(FIN) and homeostasis model assessment of insulin resistance(HOMA-IR). Thirdly, they could both improve the protein expression levels of the key signaling molecules in the IRS1/PI3K/Akt pathway and glomerulosclerosis in varying degrees, and their ameliorative effects were similar. Finally, both could improve podocyte injury and EMT, and TFA was superior to ROS. In conclusion, this study suggested that podocyte EMT and glomerulosclerosis could be induced by IR and the decreased activation of the IRS1/PI3K/Akt pathway in the kidney in DKD. Similar to ROS, the effects of TFA in inhibiting podocyte EMT in DKD were related to inducing the activation of the IRS1/PI3K/Akt pathway and improving IR, which could be one of the scientific connotations of TFA against DKD. This study provides preliminary pharmacological evidence for the development and application of TFA in the field of diabetic complications.

Effects of astragalin on the cell proliferation and cell cycle of prostate cancer cells through up-regulating miRNA-513 expression / 肿瘤研究与临床

Objective:To investigate the effects of astragalin on the cell proliferation and cell cycle of prostate cancer cell line C4-2B through up-regulating the expression of miRNA-513 (miR-513).Methods:Prostate cancer cell line C4-2B cells were taken and treated with 125 μg/L of astragalin for 48 h (astragalin group), and untreated C4-2B cells were set as the control group. The methyl thiazolyl tetrazolium (MTT) method was used to detect the proliferation ability of C4-2B cells in the two groups, and cell cycle was detected by using flow cytometry. The miRNAMap prediction software was used to predict that the targeted gene of miR-513 was the forkhead box protein R2 (FOXR2), and the dual luciferase gene reporter assay was used to verify it. Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect the relative expression levels of miR-513 and FOXR2 mRNA in the two groups of cells. Western blotting was used to detect the expressions of FOXR2, cyclin-dependent kinase 7 (CDK7), β-actin and cyclin H in the two groups of C4-2B cells.Results:Compared with the control group, the proliferation activity of C4-2B cells in the astragalin group was decreased from day 2 to day 5 (all P < 0.05). The proportions of S-phase cells in the control group and the astragalin group were (48.1±3.2)% and (36.0±2.1)%, respectively. The proportion of S-phase cells in the astragalin group was decreased ( t = 3.12, P = 0.021); the proportions of G 2-phase cells were (24.9±3.3)% and (11.8±2.4)%, respectively. The proportion of G 2-phase cells in the astragalin group was decreased ( t = 3.18, P = 0.019). The relative expression levels of miR-513 in C4-2B cells of the control group and the astragalin group were 1.01±0.22 and 6.55±0.61, respectively. The relative expression levels of miR-513 in C4-2B cells in the astragalin group was increased ( t = 7.70, P < 0.01). The dual luciferase reporter gene assay verified that FOXR2 was the targeted gene of miR-513. The relative expression level of FOXR2 mRNA in C4-2B cells of the control group and the astragalin group was 1.04±0.14 and 0.19±0.06, respectively, and the difference was statistically significant ( t = 5.53, P = 0.002), suggesting that after astragalin promoted the expression of miR-513, the FOXR2 mRNA expression was decreased. The relative expression levels of FOXR2, CDK7 and cyclin H protein in C4-2B cells in the astragalin group were all decreased compared with those in the control group. Conclusions:Astragalin inhibits the proliferation of prostate cancer C4-2B cells and induces cell cycle arrest by up-regulating the expression of miR-513.

Effects of total flavonoids of Bidens pilosa on insulin resistance in HepG 2 cells / 中国药房

OBJECTI VE To explore the effects of total flavonoids of Bidens polisa L.（TFB）on insulin resistance （IR）of HepG2 cells. METHODS B. polisa L. was refluxed and extracted with 80％ ethanol to obtain TFB. Palmitic acid was used to induce IR mode of HepG 2 cells in vitro . The effects of low-concentration ，medium-concentration and high-concentration （20，40， 80 mg/L） of TFB on the consumption of glucose were investigated. Using metformin as positive control ，the effects of low-concentration，medium-concentration and high-concentration （20，40，80 mg/L）of TFB on the protein expression of insulin receptor substrate- 1（IRS-1），c-Jun N-terminal kinase （JNK）and protein kinase C （PKC）were investigated. Molecular docking technology was used to explore the interaction between eight main active components of TFB such as quercetin ，quercitrin and IRS-1，JNK and PKC proteins. RESULTS The glucose consumption of TFB low-concentration ，medium-concentration and high-concentration groups were increased significantly （P＜0.05 or P＜0.01）. Compared with normal group ，the expression of IRS-1 and JNK protein in the model group decreased significantly ，and the expression of PKC protein increased significantly （P＜ 0.01）. Compared with model group ，the protein expression of IRS- 1 and JNK could up-regulated while the protein expression of PKC down-regulated in TFB low-concentration ，medium-concentration and high-concentration groups and metformin positive control group （P＜0.05 or P＜0.01）. The score of molecular docking energy between maritimetin in TFB and IRS- 1 protein was －7.9 kcal/mol（1 kcal＝4.816 kJ）. The scores of molecular docking energy of maritimetin ，rutin and JNK protein were －9.3 kcal/mol. The score of molecular docking energy between quercitrin and PKC protein was －4.9 kcal/mol. Interactions between components and proteins included forming hydrogen bonds ，hydrophobic bonds and so on. CONCLUSIONS TFB can significantly improve IR of HepG 2 cells，the mechanism of which may be related to the regulation of protein expression of IRS ，JNK and PKC. Maritimetin，rutin and quercitrin may be potential active ingredients for improving IR.

Chemical constituents of petroleum ether extract from the stems and leaves of Humulus scandens / 中国药科大学学报

@#To study the chemical constituents of petroleum ether extract from the stems and leaves of Humulus scandens (family of Moraceae), fifteen compounds were isolated from the stems and leaves of H.scandens by silica gel, Sephadex LH-20, ODS, and preparative HPLC chromatography.The structures were identified by physicochemical data and spectroscopic method as tectochrysin (1), chrysin (2), 5-hydroxy-3, 4'', 6, 7-tetramethoxyflavone (3), (2S)-5-hydroxy-7, 8-dimethoxyflavanone (4), imperatorin (5), phellopterin (6), ethyl 4-hydroxy-3-(3''-methyl-2''-butenyl)benzoate (7), p-hydroxy-phenylpropionic acid (8), ethyl p-hydroxycinnamate (9), p-hydroxybenzaldehyde (10), anofinic acid (11), 5,6-dehydrokavain (12), physcion (13), olean-12-ene-3,?11-dione (14) and ergosta-4, 6, 8 (14), 22-tetraen-3-one (15), respectively.All compounds were isolated from this plant for the first time.

Research progress of the chemoprevention agents for breast cancer / 国际外科学杂志

As one of the most common malignant tumors in women,breast cancer has the characteristics of high inc idence,strong invasion,easy recurrence and metastasis.Although the mortality of breast cancer has decreased in recent years,its incidence has not been effectively controlled.Chemoprevention is a kind of preventive measure to reduce the incidence of disease by using natural or synthetic chemical drugs to prevent the asymptomatic high-risk population.At present,only estrogen receptor modulators and aromatase inhibitors are allowed to be used in clinical chemoprevention of breast cancer.Therefore,researcher should actively take measures to find new highefficient,low-toxic chemoprevention agents that take into account the majority of the population.The main purpose of this review is to summarize the mechanisms and related experimental studies of six chemoprevention agents for breast cancer including estrogen receptor modulators,include aromatase inhibitors,aspirin,metformin,flavones and poly(ADP-ribose) polymerases inhibitors.

Rhizoma drynariae flavonoids inhibits rat intervertebral disc degeneration by inhibiting nuclear factor-kappa B signaling pathway / 中国组织工程研究

BACKGROUND: The combined action of mechanical compression, inflammatory factors, and matrix metalloproteinases leads to intervertebral disc degeneration. Rhizoma drynaria flavones have been shown to have anti-inflammatory effect. However, its protective effects against degeneration of the intervertebral discs remain unclear. OBJECTIVE: To investigate the potential molecular mechanisms underlying the protective effects of rhizoma drynaria flavones in a rat model of cervical intervertebral discs degeneration and analyze the clinical value. METHODS: Fifty Sprague-Dawley rats (half male and half female) were randomly divided into two groups: 10 in sham-operated group and 40 in degeneration group. Animal models of cervical intervertebral discs degeneration due to dynamic and static imbalance were made in the degeneration group. After successful modeling, 36 model rats were randomized into model group, high-dose group, middle-dose group and low-dose group (n=9 per group). Rats in the sham and model groups were intragastrically given 4 mL of normal saline per day, and those in the high-, middle- and low-dose groups were given 62.5, 125, and 250 mg/kg rhizoma drynaria solution, respectively. The C3-4, C4-5, C5-6, and C6-7 segments of the rat spine were taken. Immunohistochemistry and ELISA were used to detect expression of tumor necrosis factor α and matrix metalloproteinase 3 proteins. Quantitative RT-PCR was used to determine the mRNA expression of tumor necrosis factor α and matrix metalloproteinase 3. Western blot was used to detect the phosphorylation of IkBα. RESULTS AND CONCLUSION: Results from the immunohistochemistry, western blot, and RT-PCR, and ELISA showed that the phosphorylation of IkBα and expression of tumor necrosis factor α and matrix metalloproteinase 3 were significantly higher in the model group than the sham group (P < 0.01). However, interventions with rhizoma drynaria flavones, especially with low-dose rhizoma drynaria flavones, significantly lowered the phosphorylation of IkBα and expression of tumor necrosis factor α and matrix metalloproteinase 3. To conclude, high-, middle- and low-dose rhizoma drynaria flavones can exert protective effects against degeneration of the intervertebral discs by suppressing the nuclear factor-κB pathway and inhibiting the expressions of tumor necrosis factor α and matrix metalloproteinase 3. And low-dose rhizoma drynariae flavonoids have the optimal outcomes.

Effects of lactagogue of four total alkaloids of barley malt via dopamine D2 receptor in postpartum hypogalactia rat / 中草药

Objective: To explore the regulatory effect of malt alkaloid on prolactin (PRL) secretion in the model rat with postpartum hypogalactia induced by bromocriptine based on dopamine D2 receptor, and determine the active fraction of the malt with galactogogue effect. Methods: The postpartum hypogalactia model was established by intragastric administration of bromocriptine mesylate. After the model was successfully established, all groups were given corresponding drug treatment. The concentration of serum PRL, estradiol (E2) and progesterone (P) in each group was detected by ELISA kits. HE staining was used to observe the pathologic changes of breast tissue. RT-PCR was used to determine the mRNA levels of prolactin receptor (PRLR) and dopamine D2 receptor (DRD2) in pituitary gland of rats. Results: Compared with the control group, the levels of serum PRL, P, and E2 were significantly decreased in the model group as well as the mRNA expression of the pituitary PRL cells. But the mRNA expression of the pituitary DRD2 in the model group was significantly increased compared with the control group. Compared with the model group, the malt total alkaloid significantly increased the volume of mammary lobule and dilated the duct. There was a lot of milk in the duct and acinar in the malt total alkaloid group. Besides, the total alkaloids increased the concentration of serum PRL, P, and E2 and the mRNA expression of the pituitary PRL cells, and decreased the mRNA expression of the pituitary DRD2. Conclusion: The primary the active fraction of malt for galactogogue action is total alkaloids, and its mechanism may be related to promoting PRL secretion, increasing serum PRL receptor level and decreasing the mRNA expression of dopamine D2 receptor.

Constituent analysis of Jingzhi Guanxin Soft Capsule based on UHPLC-Q-Orbitrap HRMS / 中草药

Objective: In order to establish a rapid and efficient analysis method for identification of the complex components in Jingzhi Guanxin Soft Capsule, and provide the basic research data for the systematic elaboration of its chemical constituents. Methods: An ultra-high performance liquid chromatography-quadrupole/Orbitrap high resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) was used for the identification analysis of the components in Jingzhi Guanxin Soft Capsule, and the multistage fragments ions data was compared with the standard substance and literature consulting. Results: Forty-three compounds were identified in this study, including flavones, phthalides, organic acids, quinones and other categories. Conclusion: The chemical constituents of Jingzhi Guanxin Soft Capsule are identified systematically, accurately and efficiently, which provide the theory basis for the further research of its pharmacodynamic material basis and quality control.

Research progress of the chemoprevention agents for breast cancer / 国际外科学杂志

As one of the most common malignant tumors in women, breast cancer has the characteristics of high incidence, strong invasion, easy recurrence and metastasis. Although the mortality of breast cancer has decreased in recent years, its incidence has not been effectively controlled. Chemoprevention is a kind of preventive measure to reduce the incidence of disease by using natural or synthetic chemical drugs to prevent the asymptomatic high-risk population. At present, only estrogen receptor modulators and aromatase inhibitors are allowed to be used in clinical chemoprevention of breast cancer. Therefore, researcher should actively take measures to find new high-efficient, low-toxic chemoprevention agents that take into account the majority of the population. The main purpose of this review is to summarize the mechanisms and related experimental studies of six chemoprevention agents for breast cancer including estrogen receptor modulators, include aromatase inhibitors, aspirin, metformin, flavones and poly(ADP-ribose) polymerases inhibitors.

Content determination of total flavones and polysaccharides in Fangshu Qingre mixture by UV-Vis spectrophotometry / 药学实践杂志

Objective To establish a method for content determination of total flavones and polysaccharides in Fangshu Qingre mixture by UV-Vis Spectrophotometry. Methods The contents of total flavones and polysaccharides in Fangshu Qingre mixture were determined by UV-Vis spectroscopy with rutin and anhydrous glucose as reference substance, and the wavelength was set at 508 nm and 487 nm. Results The contents were from 0.00 to 59.20 μg/ml for total flavones and from 10.92 to 109.20 μg/ml for total polysaccharides in Fangshu Qingre mixture. The recoveries of total flavones and total polysaccharides were 104.4% and 104.8% respectively. Conclusion The method of using ultraviolet spectroscopy was simple, reproducible, accurate and reliable, which could be preferably used as the method for content determination of total flavones and polysaccharides in Fangshu Qingre mixture.

Rapid Identification on Chemical Components in Dalbergiae Odoriferae Lignum by UPLC-Q-TOF-MS/MS / 中国实验方剂学杂志

Objective::Dalbergiae Odoriferae Lignum is a rare traditional Chinese medicine material in China. However, there are many varieties of various sources and different qualities in the market at present. In order to further define the pharmacodynamic substance basis, electrospray time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS) was used to rapidly analyze chemical constituents of methanol extract of Dalbergiae Odoriferae Lignum. Method::Chromatographic separation was performed on an UPLC RRHD SB-C18(3.0 mm×100 mm, 1.8 μm)for gradient elution, with mixtures of acetonitrile and 0.1%formic acid-water as mobile phases at a flow rate of 0.3 mL·min-1. The column temperature was maintained at 40 ℃. The data was collected in a negative ion mode with electro-spray ionization source(ESI). Result::According to molecular ion peaks and MS2 mass spectrometry characteristic fragment ions, Mass Bank databases, as well as the mass spectrometry information of reference substances and relevant literatures, a total of 83 constituents were identified, including 18 flavones, 31 isoflavones, 10 neoflavonoids, 9 isoflavanones, 7 other flavonoids and 8 other components. Conclusion::UPLC-Q-TOF-MS/MS can quickly, accurately and comprehensively identify chemical constituents in methanol extract of Dalbergiae Odoriferae Lignum, and isoflavones, flavones, neoflavonoids and isoflavanones are the main chemical constituents, which laid a foundation for the basic research of medicinal substances of Dalbergiae Odoriferae Lignum, and provided theoretical basis and technical support for the improvement of quality standards of Dalbergiae Odoriferae Lignum.

Multiple constituents study of Qianliexin Capsule based on UHPLC-Q-Orbitrap HRMS / 中草药

Objective: In order to establish an efficient analysis method for the identification of the complex components in Qianliexin Capsule rapidly, and provide the basic research data for the systematic elaboration of its chemical constituents. Methods: An ultra-high performance liquid chromatography-quadrupole/Orbitrap high resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) was used for the identification analysis of the components in Qianliexin Capsule accurately, and the multistage fragments ions data was compared with the standard substance and consulting the literatures. Results: Sixty-one compounds were identified in this study, including 12 flavones, 11 coumarins, six terpenes, 19 organic acids and other categories. Conclusion: The accurate and rapid identification of various chemical constituents of Qianliexin Capsule was achieved in this study, which provided the theory basis for the pharmacodynamic material and the quality control study of Qianliexin Capsule.

Study on chemical constituents of Shensong Yangxin Capsule based on UHPLC-Q-Orbitrap HRMS / 中草药

Objective: To investigate the chemical composition of Shensong Yangxin Capsule, a novel ultra high performance liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) was employed to establish a method that could identify the ingredients systematically and rapidly. Methods: The information of accurate mass and multistage fragment ions was obtained by the novel UHPLC-Q-Orbitrap technology. Chemical constituents were characterized by comparing the relative retention time and the mass data of the reference substance, meanwhile consulting the reference literature or the Mass Bank, Chemical Book network database. Results: A total of 54 compounds were finally identified in this research, including the phenolic acids, flavones, terpenoids, anthraquinones, alkaloids and others. Conclusion: A method was established in this study to identify various chemical constituents of Shensong Yangxin Capsule systematically, rapidly and accurately. The identified chemical components mostly cover the main constituents of each medicinal material in the formula. Our work will lay a scientific foundation for the bioactive components screening, quality control improvement and further clinical application of this herbal formulation.

Effect of Rosae Chinensis Flos Total Flavones on Focal Cerebral Ischemia-reperfusion in Rats / 中国实验方剂学杂志

Objective: To study the effect of Rosae Chinensis Flos total flavones(RCTF) on the focal cerebral ischemia-reperfusion model in rats, in order to preliminarily explore the mechanism of action. Method: Rats were randomly divided into sham-operated group, model group, large, medium, and low-dose RCTF group(200,100,50 mg ·kg-1) and positive group[Nimodipine group(20 mg ·kg-1) and Naoluotong group (500 mg ·kg-1)]. After 7 days of continuous administration, 1 hour later after the last administration, the middle cerebral artery middle cerebral artery occlusion (MCAO) model was duplicated. After 2 hours of modeling, perfusion was performed for 22 hours. Mortality and neurological deficits were scored. Serum S-100β was detected; brain tissue malondialdehyde(MDA), superoxide dismutase (SOD), nitric oxide (NO), nitric oxide synthase (NOS), tumour necrosis factor-α(TNF-α), interleukin-1β(IL-1β), intercellular adhesion molecule-1(ICAM-1), adenosine triphosphate (ATP)ase were measured. The brain tissue morphological changes were observed. Result: The rat model of focal cerebral ischemia and reperfusion was successfully replicated. Compared with the model group, RCTF in large, medium, and low-dose RCTF group significantly decreased the score of neurological deficit in rats (Pβ in serum (PPP+K+-ATPase, Mg2+-ATPase, and Ca2+ in brain tissue (Pα content, IL-1β, ICAM-1 content in brain tissue (PPConclusion: RCTF have a protective effect on cerebral ischemia-reperfusion injury in rats. The mechanism may be related to the resistance of anti-free radicals, the reduction of inflammation in brain tissue and the improvement of brain energy metabolism after cerebral ischemia reperfusion injury.

Study on Protective Effect of Total Flavonoids from the Leaves of Mongolian Medicine Choerospondias axillaris on Myocar- dial Ischemia Reperfusion Injury Model Rats / 中国药房

OBJECTIVE： To investigate the protective effect of total flavonoids from the leaves of Choerospondias axillaris （TFLC） on myocardial ischemia reperfusion injury （MIRI） model rats. METHODS： Male SD rats were randomly divided into sham operation group， model group， positive control group （verapamil， 0.02 g/kg）， TFLC low-dose and high-dose groups （0.1， 0.4 g/kg）， with 10 rats in each group. Administration groups were intragastrically given relevant medicine （2 mL/100 g）； sham operation group and model group were given constant volume of normal saline intragastrically， once a day， for consecutive 7 d. After last medication， MIRI model was induced by modified ligation method. The times and duration of ventricular tachycardia （VT） and ventricular fibrillation （VF） in rats were recorded with biological function experiment system during reperfusion period.The activity of CK and contents of TNF-α， IL-6， NF-κB and NO in serum were determined by ELISA double antibody clip art assay. The morphological characteristics of myocardial tissue was observed by HE staining. The myocardial infarction scope （i.e. the ratio of myocardial tissue mass to ventricular mass） was measured by TTC method. RESULTS： Compared with sham operation group， the times and duration of VT and VF were increased or prolonged significantly in model group； CK activity， serum contents of TNF-α， IL-6 and NF-κB were enhanced or increased significantly， while NO content was decreased significantly （P＜0.05 or P＜0.01）. Obvious myocardial infarction focus， serious cell structure damage， disorderly muscle fibers arrangement， cell nucleus pyknosis and accompanied inflammatory cell infiltration were all observed in cardiac tissue； the mass of infarcted myocardial tissue and ventricular as well as the scope of myocardial infarction increased significantly （P＜0.01）. Compared with model group， the times and duration of VT and VF were decreased or shortened significantly in administration groups； CK activity， serum contents of TNF-α， IL-6， NF-κB were decreased significantly， while NO content was increased significantly （P＜0.05 or P＜0.01）. The above symptoms of myocardial injury were improved；the mass of infarcted myocardial tissue and ventricular as well as the scope of myocardial infarction was significantly reduced （P＜0.05 or P＜0.01）. CONCLUSIONS： TFLC can relieve MIRI-induced ischemic arrhythmia and myocardial damage， reduce the release of inflammatory factors， promote the recovery of myocardial and endothelial cell function， reduce the scope of myocardial infarction and has a certain protective effect.

Sophoral flavones inhibit proliferation of cardiac fibroblasts via regulating the expression of prohibitin / 药学学报

The aim of the present study was to investigate the effect of Sophoral flavones on proliferation of cardiac fibroblasts (CFb) induced by high glucose and its underlying mechanism. Cardiac fibroblasts were exposed to different concentration of D-glucose (15, 25 and 35 mmol·L-1) at different time point (24, 48 and 72 h) in order to determine cell proliferation, and the model group was established by culturing CFb with 25 mmol·L-1D-glucose for 48 h. Sophoral flavones (12.5, 25 and 50 mg·L-1) were employed for intervention. The cell viability was measured by MTT assay, and the levels of transforming growth factor-β1 (TGF-β1), matrix metalloproteinase-2 (MMP-2), collagen Ⅰ and collagen Ⅲ were measured by ELISA. In addition, flow cytometry was employed to detect the cell cycle; while the protein expression of prohibitin (PHB) was observed via immunocytochemistry and Western blot. This animal experiment had been approved by Jilin Medical University Experiment Animal Ethics Review Committee. The results showed that 25 mmol·L-1 glucose could promote the proliferation of CFb; and the contents of TGF-β1, MMP-2, collagen Ⅰ and collagen Ⅲ in the model group were higher than that of control (P<0.05). The number of cells in S and G2 phase increased under high glucose condition. In the model group, PHB translocation occurred at 6 h and protein expression decreased at 48 h (P<0.01). Compared with the model group, 12.5-50 mg·L-1 Sophoral flavones reduced the contents of TGF-β1, MMP-2, collagen Ⅰ and collagen Ⅲ, increased the number of G1 phase cells, and increased the expression of PHB protein at 48 h (P<0.05), with no effect on the nuclear translocation of PHB. These results indicated that Sophoral flavones could prevent the proliferation of CFb induced by high glucose, the mechanism of which may be related to increasing the expression of PHB protein.

Effects of total flavones of rhododendra on contractility of rat myocardial cells and its mechanism / 安徽医科大学学报

Objective To investigate the total flavones of rhododendra(TFR) on contractility of rat myocardial cells and its possible mechanism. Methods The contraction amplitude and contraction frequency of primary cultured rat myocardial cells were observed by image analysis system. The intracellular free Ca2 + content was measured by calcium ion imaging system. Results 10 and 100 nmol /L hU Ⅱ significantly accelerated the contraction frequency of myocardial cells,and 10 nmol /L hU Ⅱ increased the contraction amplitude of myocardial cells,but 100 nmol /L hU Ⅱ reduced the contraction amplitude of myocardial cells. TFR 300 mg /L significantly slowed the contraction frequency of rat myocardial cells and increased the contraction amplitude. TFR in the range of 33. 3 ～ 300 mg /L could significantly inhibit the increase of contraction frequency,the decrease of contraction amplitude and the increase of intracellular free Ca2 + content induced by 100 nmol /L hU Ⅱ. Conclusion TFR can slow down the contraction frequency of myocardial cells and increase its contractility,which may be related to the decrease of free Ca2 + content in myocardial cells.

Comparison of bioactive compounds content in leaf extracts of Passiflora incarnata, P. caerulea and P. alata and in vitro cytotoxic potential on leukemia cell lines

ABSTRACT Passiflora caerulea L., P. alata Curtis and P. incarnata L. (synonym for P. edulis Sims), are the most popular representatives of the Passiflora genus in South America. In recent years, a growing attention is paid to the biological activity and phytochemical profiles of crude extracts from various species of Passiflora in worldwide. The aim of this study was to evaluate and to compare of anti-leukemic activity of the dry crude extracts from leaves of three Passiflora species from greenhouse of Poland in two human acute lymphoblastic leukemia cell lines: CCRF-CEM and its multidrug resistant variant. Two systems of liquid chromatography in order to assessment of phytochemical composition of extracts were applied. Extracts of P. alata and P. incarnata showed the potent inhibitory activity against human acute lymphoblastic leukemia CCRF-CEM, while P. caerulea not showed activity (or activity was poor). Despite similarities in quality phytochemical profile of extracts from P. caerulea and P. incarnata, differences in quantity of chemical compounds may determine their various pharmacological potency. For the activity of P. alata extract the highest content of terpenoids and a lack of flavones C-glycosides are believed to be crucial. Summarizing, the crude extract from P. alata leaves may be considered as a substance for complementary therapy for cancer patients.