Study on quality standard of Yi medicine Gynura japonica and quality evaluation of medicinal materials from different producing areas / 中国药房

OBJECTIVE To improve the quality standard of Yi medicine Gynura japonica， and to evaluate its quality. METHODS Using 15 batches of G. japonica from different producing areas as samples， the contents of water， total ash， acid- insoluble ash and water-soluble extract were determined according to the method stated in part Ⅳ of Chinese Pharmacopoeia （2020 edition）. The contents of total alkaloid （calculated by senecionine） was determined by UV spectrophotometry. The contents of senecionine and seneciphylline were determined by HPLC. Using above 7 indexes as evaluation indexes， cluster heat map analysis， principal component analysis （PCA） and entropy weight approximation ideal ranking （TOPSIS） were used to evaluate the quality of medicinal material comprehensively. RESULTS Among 15 batches of G. japonica， the moisture contents were 8.88%-12.60%， the total ash contents were 4.43%-11.02%， the acid-insoluble ash contents were 0.56%-3.45%， the water-soluble extract contents were 21.71%-53.91%， the total alkaloid contents （calculated by senecionine） were 0.15%-0.39%， and the contents of senecionine and seneciphylline were 0.01% -0.05% and 0.01%-0.06% respectively. According to the results of various indicators， it was preliminarily proposed that the water content in the sample of G. japonica should not exceed 13.00%， the total ash content should not exceed 11.50%， the acid-insoluble ash content should not exceed 3.70%， the water-soluble extract should not be less than 20.70%， the total alkaloid content should not be less than 0.15%， the contents of senecionine and seneciphylline should not be less than 0.01% both. The results of cluster heat map analysis showed that the 15 batches of samples could be divided into four categories； the results of PCA and TOPSIS showed that the samples with high-quality ranking were jsq-2， jsq-5， jsq-6 and jsq-10， and the samples with low-quality ranking were jsq-4， jsq-13 and jsq-14. CONCLUSIONS In this study， the quantitative analysis method of total alkaloids （calculated by senecionine）， senecionine and seneciphylline in G. japonica is established， and the limits of each index are preliminarily determined. Among 15 batches of samples， the qualities of medicinal material collected from Linza Village of Ganluo County of Liangshan Yi Autonomous Prefecture， Machangping Village of Luojishan Town of Puge County of Liangshan Yi Autonomous Prefecture and other places are better.

Antitumor effect and mechanism of total alkaloids of Gelsemium elegans and sempervirine in vitro / 中国药房

OBJECTIVE To explore the antitumor effect and mechanism of total alkaloids of Gelsemium elegans （TA） and sempervirine （SPV） in vitro. METHODS The effects of low， medium and high concentrations of TA （50， 100， 200 μg/mL） and SPV （10， 30， 50 μmol/L） on the morphology of human hepatoma cells （HepG2， Bel-7402）， human lung cancer cells （A549） and human colon cancer cells （HCT-8） were observed， and the toxicity of TA and SPV to four tumor cells was monitored. The effects of TA and SPV on the contents of caspase-3 and caspase-9 in the supernatant of HCT-8 cells， the protein expressions of phosphorylated protein kinase B （p-Akt） （Thr308， Ser473）， B-cell lymphoma 2 （Bcl-2）， Bcl-2-related X protein （Bax）， survivin， C/EBP-homologous protein （CHOP）， immunoglobulin binding protein （Bip） and microtubule-associated protein 1 light chain 3Ⅱ （LC3Ⅱ） in HCT-8 cells were detected. RESULTS After the intervention of TA and SPV， the volume reduction and nuclear shrinkage were founded in four tumor cells； the cell activity decreased to varying degrees， among which TA and SPV had the best inhibitory effect on HCT-8 cells. After the intervention of TA and SPV， the contents of caspase-3 and caspase-9 in the supernatant of HCT-8 cells， the protein expressions of Bax， CHOP， Bip and LC3Ⅱ all increased to different degrees， while the protein expressions of p-Akt （Thr308， Ser473）， Bcl-2 and survivin in HCT-8 cells all decreased to different degrees. CONCLUSIONS TA and SPV have inhibitory effects on the above four tumor cells， and the inhibitory effect on HCT-8 cells is the best. The mechanism of their action on HCT-8 cells may be related to promoting apoptosis， activating endoplasmic reticulum stress and autophagy.

Study on antidepressant effect of total alkaloids of Fibraurea recisa / 中国中药杂志

This study aimed to investigate the antidepressant effects of total alkaloids of Fibraurea recisa in HT22 cells damaged by corticosterone (CORT) in vitro and in a mouse model of chronic unpredictable mild stress (CUMS) as well as the underlying mechanisms.In cellular experiments,the viability of CORT-damaged HT22 cells was detected using cell counting kit-8 (CCK-8),and the cell apoptosis was detected by Hoechst 33258 staining.In animal experiments,C57BL/6N mice were randomly divided into the control group,model group,low (100 mg·kg~(-1)),medium (200 mg·kg~(-1)) and high (400 mg·kg~(-1))-dose of total alkaloids of F.recisa groups,and positive control group.After 21 days of CUMS exposure,their depressive behaviors were observed in behavioral and Morris water maze tests.The serum levels of 5-hydroxytryptamine (5-HT),dopamine (DA),and norepinephrine (NE) were assessed by ELISA.The expression levels of apoptosis-related proteins Bcl-2,Bax and cleaved caspase-3 in HT22 cells and mouse hippocampus were detected by Western blot.The results suggested that total alkaloids of F.recisa alleviated the damage of HT22 cells induced by CORT in a dose-dependent manner.The Hoechst 33258 staining uncovered that total alkaloids of F.recisa better reduced the blue spots and inhibited cell apoptosis.The results of animal experiments showed that total alkaloids of F.recisa significantly improved the depression-like behaviors of mice and increased the serum levels of 5-HT,DA and NE as compared with those in the model group.The Western blot assays revealed a significant up-regulation of Bcl-2 protein expression,but an obvious reduction in Bax and cleaved caspase-3protein expression in the total alkaloids of F.recisa group.In conclusion,total alkaloids of F.recisa inhibited depression possibly by regulating the apoptosis-related protein expression or elevating the monoamine neurotransmitter levels in the brain.

Effect of capparis spinosa total alkaloid on proliferation and apoptosis of nucleus pulposus cells in an intervertebral disc degeneration rat model / 中国组织工程研究

BACKGROUND: Capparis spinosa total alkaloids (CSTA) have certain effects on cell growth and extracellular matrix synthesis. The aging and apoptosis of nucleus pulposus cells are one of the main pathologies of intervertebral disc degeneration. Therefore, it is assumed that CSTA may have certain effect on the degeneration of the intervertebral disc. OBJECTIVE: To study the effect of CSTA on intervertebral disc degeneration rat model and nucleus pulposus cells. METHODS: Thirty-two Sprague-Dawley rats were randomly divided into sham, model, CSTA-H, and CSTA-L groups with eight in each group. Rat models of intervertebral disc degeneration were made in the model group, CSTA-L group and CSTA-H group. The CSTA-L and CSTA-H groups were given intragastric administration of CSTA 225 mg/kg/d and 450 mg/kg/d for 4 weeks respectively. Hematoxylin-eosin staining was used to observe the pathological changes of the intervertebral disc. Immunohistochemistry and western blot were used to detect the expression of type II collagen and aggrecan. Nucleus pulposus cells from the intervertebral disc of another two Sprague-Dawley rats were separated, cultured and divided into a control group, an oxygen-glucose deprivation (OGD) group and an administration group (OGD+CSTA 10 mg/L). After being cultured for 24 hours, the morphology of nucleus pulposus cells was observed, the cell proliferation ability was detected by cell counting kit-8, the cell apoptosis was detected by flow cytometry, and the expression of type II collagen and aggrecan were detected by western blot. RESULTS AND CONCLUSION: (1) Compared with the sham group, the intervertebral disc tissue of the model group showed fiber ring fissures, aggregation and shrinking of nucleus pulposus cells, and different improvements were found in the CSTA-L and CSTA-H groups. (2) The expression levels of type II collagen and aggrecan in the model group were significantly lower than those in the sham, CSTA-L and CSTA-H groups (P < 0.05). (3) Compared with the control group, the cells in the OGD group showed irregular morphology and death status, whereas the cell morphology in the administration group was improved. (4) Compared with the control group, nucleus pulposus cells in the OGD group showed lower proliferation, higher apoptotic rate, and lower levels of type II collagen and aggrecan (P < 0.05). Compared with the OGD group, nucleus pulposus cells in the administration group showed faster proliferation, lower apoptotic rate, and higher levels of type II collagen and aggrecan. To conclude, CSTA can improve intervertebral disc degeneration by promoting proliferation and inhibiting apoptosis of nucleus pulposus cells as well as inhibiting the degradation of extracellular matrix.

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.

Effect of Total Alkaloids from Nandina domfestica in Attenuating Arsenic Trioxide and Its LC-MS / 中国实验方剂学杂志

Objective::To study the effect of total alkaloids of Nandina domestica in attenuating arsenic trioxide, and analyze chemical constituents of alkaloids extract of Nandina, in order to provide the theoretical basis for studying the effect of N. domestica in attenuating arsenic trioxide with alkaloids extract of N. domestica as effective fraction. Method::The model of acute liver injury induced by arsenic trioxide was used to compare the effects on heart and liver functions of mice between arsenic trioxide alone and total alkaloids of N. domestica combined with arsenic trioxide. The detoxification of total alkaloids on arsenic trioxide was evaluated based on biochemical parameters and pathological report. Peakview (Version1.2, AB SCIEX) software was used to process the mass spectrometry data of total alkaloids of N. domestica. The structures of determined compounds were identified by molecular weight of compound (molecular formula), secondary fragments, chromatographic peak retention and literature information. Result::Among biochemical indicators, creatine kinase(CK), lactate dehydrogenase(LDH), blood urea nitrogen(BUN), malondialdehyde(MDA), alanine aminotransferase(ALT) and amino transferase of aspartate(AST) of the arsenic trioxide group were increased, while elimination rates of Na+ -K+ -adenosine triphosphate(ATP), superoxide dismutase(SOD), catalase(CAT) and serum creatinine(SCr) were decreased, compared with those of the combination group. CK and LDH of the alkaloids extract group were more obviously increased than those of the N. domestica extract group, but with no remarkable difference. In histomorphometric examination, edema of mouse heart cells was improved, and some kidney and liver damages in rats were alleviated. Totally 25 alkaloids of alkaloid extract were identified. Among them, 18 were known, and 7 were unknown, including 3 structural types, in which apomorphine alkaloids were mostly. Conclusion::Heart, kidney and liver damage degrees of the combination group were significantly alleviated compared with the arsenic trioxide group. The total alkaloids fraction extracted and purified have a significant attenuation in arsenic trioxide toxicity. The detoxification of total alkaloids extract was equal to that of N. domestica extract. Furthermore, apomorphine alkaloids, such as nantenine and domesticine, can be used as index components to establish a quality control method for total alkaloids.

Total alkaloids of Sophora alopecuroides- and matrine-induced reactive oxygen species impair biofilm formation of Staphylococcus epidermidis and increase bacterial susceptibility to ciprofloxacin / 中草药·英文版

Objective: To investigate the mechanism by which total alkaloids of Sophora alopecuroides (TASA) and matrine (MT) impair biofilm to increase the susceptibility of Staphylococcus epidermidis (S. epidermidis) to ciprofloxacin. Methods: The minimum biofilm inhibitory concentration (mBIC) was determined using a 2-fold dilution method. Structure of biofilm of S. epidermidis was examined by Confocal Laser Scanning Microscope (CLSM). The cellular reactive oxygen species (ROS) was determined using a DCFH-DA assay. The key factors related to the regulation of ROS were accessed using respective kits. Results: TASA and MT were more beneficial to impair biofilm of S. epidermidis than ciprofloxacin (CIP) (P < 0.05). TASA and MT were not easily developed resistance to biofilm-producing S. epidermidis. The mBIC of CIP decreased by 2–6-fold following the treatment of sub-biofilm inhibitory concentration (sub-BIC) TASA and MT, whereas the mBIC of CIP increased by 2-fold following a treatment of sub-BIC CIP from the first to sixth generations. TASA and MT can improve the production of ROS in biofilm-producing S. epidermidis. The ROS content was decreased 23%−33% following the treatment of sub-mBIC CIP, whereas ROS content increased 7%−24% following treatment with TASA + CIP and MT + CIP combination from the first to sixth generations. Nitric oxide (NO) as a ROS, which was consistent with the previously confirmed relationship between ROS and drug resistance. Related regulatory factors-superoxide dismutase (SOD) and glutathione peroxidase (GSH) could synergistically maintain the redox balance in vivo. Conclusion: TASA and MT enhanced reactive oxygen species to restore the susceptibility of S. epidermidis to ciprofloxacin.

Isolation and Identification of Total Alkaloids from Mongolian Medicine Aconitum excelsum and Its Therapeutic Mechanism on Chronic Bronchitis in Rats / 中国药学杂志

OBJECTIVE: To isolate and identify the total alkaloids from Aconitum excelsum and to study the therapeutic mechanism of the total alkaloids from Aconitum excelsum on chronic bronchitis in rats. METHODS: The total alkaloids from Aconitum excelsum was isolated by TLC and the structures of compounds were elucidated by a variety of spectral techniques. The rat model of chronic bronchitis (CB) was established with combined utilization of bacillus calmette-guerin and lipopolysaccharide, dexamethasone tablets and total alkaloids from Aconitum excelsum (high and low dose) were ig administrated. The pathological changes of lung tissue in rats were observed by HE and the level changes of interleukin-1 (IL-1), interleukin-8 (IL-8) and tumor necrosis factor-α (TNF-α) in serum were determined by ELISA method. RESULTS: Seven alkaloids were identified from the total alkaloids of Aconitum excelsum. The administration of total alkaloids from Aconitum excelsum could decrease the degree of infiltration, epithelial cell shedding, bronchial secretion, inhibit the proliferation of bronchial epithelial cells of the rat bronchial inflammatory cells, and make the bronchial diameter increases and ventilation. The total alkaloids could significantly inhibit the increase of IL-1, IL-8 and TNF-α in the serum of rats with CB, compared with the model group with the statistical significance (P<0.05). CONCLUSION: The total alkaloids from Aconitum excelsum have the better treatment effect on CB, which may be related to its main chemical components, namely lappaconitine (1), ranaconitine (2), septentriod-ine (3), 6-demethyldelsodine (4), 6-methylumbrofine (5), 8-methyl-10-hydroxylycoctonine (6) and 8-methyllycoctonine (7).

Study on mechanism of total alkaloids of Sophora flavescens against Staphylococcus epidermidis / 中草药

Objective: To detect the effect of alkaloids of Sophora flavescens on biofilm formation in Staphylococcus epidermidis in vitro, and to observe whether total alkaloids of S. flavescens can affect the tolerance of Staphylococcus epidermidis to lactic acid and hydrogen peroxide, then explore the possible anti-microbial mechanism of alkaloids in Sophora flavescens. Methods: The biofilm of Staphylococcus epidermidis was prepared in vitro. We evaluated the effect of alkaloids in S. flavescens on the biofilm-forming ability of Staphylococcus epidermidis via 96-well cell culture microtiter plates with crystal violet staining as well as Congo red experiment. The biofilm formation of Staphylococcus aureus was observed using scanning electron microscope. Then, we measured the change of tolerance to oxidative stress and sensibility to antibiotics for Staphylococcus epidermidis being treated by alkaloids in S. flavescens Ait. qRT-PCR analysis was performed to show the relative transcript level of serp2195 and gpxA-2 upon Staphylococcus epidermidis strain exposed to alkaloids in S. flavescens at different concentrations. Results: Alkaloids in S. flavescens significantly prevented biofilm formation in Staphylococcus epidermidis (P < 0.001) at the concentration of 5.0 mg/mL during crystal violet staining and Congo red experiment, and thus weakened tolerance to oxidative stress and enhanced sensibility to lactate for Staphylococcus epidermidis. Alkaloids in S. flavescens could downregulate the transcript level of serp2195 and gpxA-2. Conclusion: Alkaloids in S. flavescens has a significant inhibitory effect on biofilm formation of Staphylococcus epidermidis, which weakens the tolerance of bacteria to oxidative stress. The mechanism may be realized by downregulating the transcript level of serp2195 and gpxA-2, and alkaloids in S. flavescens could reduce Staphylococcus epidermidis tolerance to lactic acid. To sum up, Alkaloids in S. flavescens can be applied to the gynecological infection caused by biofilm-producing bacteria.

Optimization of Purification Process of Total Alkaloid Extract of Berberis dictyophylla Cortex by Macroporous Resin and Investigation of Its Quality Standard / 中国实验方剂学杂志

Objective: To optimize purification process of total alkaloid extract of Berberis dictyophylla cortex by macroporous resin,and to establish its quality standard. Method: Acid dye colorimetry was used to investigate the purification process of total alkaloid extract of B. dictyophylla cortex,the process parameters included concentration of sample solution,speed of sampling,diameter-height ratio of resin column,water washing amount,concentration and dosage of eluent,flow rate of elution,etc.In order to determine the optimum process,HPLC was employed to determine the contents of four alkaloids(magnoflorine,jatrorrhizine hydrochloride,palmatine hydrochloride,and berberine hydrochloride) with mobile phase of acetonitrile-0.1% phosphoric acid aqueous solution for gradient elution and detection wavelength at 270 nm.After being purified,quality standard of total alkaloid extract of B. dictyophylla cortex was investigated according to the requirements in the 2015 edition of Chinese Pharmacopoeia. Result: Optimal purification conditions were as following:10 g of HPD100 macroporous adsorption resin with a column diameter-height ratio of 1:8,sampling solution concentration of 11 g·L-1,the loading flow rate of 1 mL·min-1,sampling solution volume of 50 mL,washed with 4 BV of water(1 BV=15 mL) and added 9 BV of 30% ethanol,after being purified,the transfer rate of total alkaloids was>80%,and its purity was>65%.The quality standard of total alkaloid extract of B. dictyophylla cortex was established,there were 19 common peaks in the characteristic chromatogram,and the overall similarity was>0.99. Conclusion: This optimized purification process is stable and feasible, and the established quality standard is controllable.

Screening and Verification of Compatibility Proportion of Effective Components from Traditional Chinese Medicine of Diabetes / 中国实验方剂学杂志

Objective: To screen the best compatibility proportion of total alkaloids of Coptidis Rhizoma and Notoginseng Radix et Rhizoma total saponins (HS composition)by uniform design and pharmacological model and to observe the effect on diabetic complications. Method: The total alkaloids of Coptidis Rhizoma and Notoginseng Radix et Rhizoma total saponins were used as the research objects, U6(62) table was choosed for grouping design.The content of triglyceride fasting blood-glucose (FBG), prothrombin time (PT) and active partial thromboplastin time (APTT) was chosen as index. The best dose ratio was obtained by multipleregression analysis. Rats with type 2 diabetes mellitus induced by high-fat diet combined streptozotocin were divided into blank group, model group, metformin group (150 mg·kg-1), HS composition group (total alkaloids of Coptidis Rhizoma 360 mg·kg-1+ Notoginseng Radix et Rhizoma total saponins 40 mg·kg-1). Rats were administered orally for 10 weeks.By observing the blood glucose, glucose tolerance,area under the curve (AUC), triglyceride (TG), high-density lipoprotein (HDL-C)and low-density lipoprotein (LDL-C), hemorheological indexes and pathological changes of pancreas, heart, kidney and retina in rats of each group, the effect of this composition on diabetic complications was verified. Result: Combination of 625 mg·kg-1 total alkaloids of Coptidis Rhizoma and 60 mg·kg-1 Notoginseng Radix et Rhizoma total saponins was the optimal dosage ratio of HS composition.The validation test showed that compared with blank group, the fasting blood glucose and lipid levels in the model group were significantly increased (PPPPPPPPPPPConclusion: The optimum compatibility dose of HS composition have a good therapeutic effect on diabetic complication rats induced by high-fat diet and streptozotocin.

Comparison of Improvement Effects and Its Mechanism of Different Doses of Total Alkaloids from Aconitum racemulosum on CIA Model Rats Based on Information Entropy Theory / 中国药房

OBJECTIVE： To investigate the effects of different doses of total alkaloids from Aconitum racemulosum （ARTA） on serum inflammation factors and FOS protein expression in synovial tissue of joint in collagen-induced arthritis （CIA） model rats， and to investigate its potential mechanism of anti-rheumatoid arthritis （RA）. METHODS： Male SD rats were randomly divided into blank group， model group， positive group （Compound dexamethasone acetate ointment， 0.2 g/kg）， ARTA low-dose， medium-dose and high-dose groups （56.26， 112.50， 225.00 mg/kg， by the weight of ARTA in the extract）， with 10 rats in each group. Except for blank group， other groups were given subcutaneous injection of Bovine collagen Ⅱ emulsified with incomplete Freund’s adjuvant into the left foot to establish CIA model； the left foot were smeared with relevant medicine from the day of modeling. Blank group and model group were smeared with constant volume of 65％ ethanol， 3 times a day， for consecutive 28 days. On the 7th， 14th， 21st and 28th day of administration， the thickness of left hind toe was measured with vernier caliper， and the degree of foot swelling was calculated. The serum contents of IL-1β， IL-6 and TNF-α in rats were measured by ELISA after last administration. The expression of FOS protein in synovial tissue was determined by immunohistochemical method [expressed by HIS]. The comprehensive score was conculated by entropy weight method. Effects of each dosage on above indexes of CIA model rats were evaluated with the comprehensive score. RESULTS： Compared with blank group， the degree of foot swelling， serum content of inflammatory factors and HIS value were increased significantly in model group （P＜0.05）. Compared with model group， the degree of foot swelling in each administration group， serum contents of IL-1β， IL-6 and TNF-α， HIS in positive group and ARTA high-dose group， serum contents of IL-6 and TNF-α in ARTA medium-dose group as well as serum content of TNF-α in ARTA low-dose group were decreased significantly（P＜0.05）. Comprehensive score of above indicators were 0.37（positive group）， 0.31（ARTA high-dose group）， 0.23（ARTA medium-dose group） and 0.09（ARTA low-dose group）. CONCLUSIONS： ARTA can improve CIA model rats， and the effect tends to increase with the increase of dose. Above effect may be associated with reducing serum content of inflammatory factors and inhibiting the expression of FOS protein in synovial tissue.

Total alkaloids of Coptidis Rhizoma combined with exercise inhibits tumor growth of orthotopically transplanted 4T1 breast cancer mice by blocking cell cycle G_1/S transformation / 中国中药杂志

Breast cancer is one of the leading causes for cancer-related death among women worldwide. Coptidis Rhizoma has antibacterial,anti-inflammatory,anti-tumor and other pharmacological activities,but whether exercise could synergistically promote the role of RC in the treatment of breast cancer has not been reported. In this experiment,the effects and mechanism of total alkaloids of Coptidis Rhizoma combined with exercise on the tumor growth of orthotopically transplanted 4 T1 breast cancer were systemically studied in mice. Balb/C mice transplanted with 4 T1 cells in situ were used as models. The total alkaloids of RC(145 mg·kg-1·d-1) alone or in combination with exercise(10 m·min-1,30 min/time,5 times/week) were given for 28 days,and then the changes in body weight and tumor volume,tumor weight,interleukin-1β(IL-1β),serum estradiol(E2) content,and expression levels of estrogen receptor α(ERα),cell cycle related proteins CDK4,CDK6,cyclin D1,CDK2,and cyclin E in tumor tissues. The results showed that total alkaloids of Coptidis Rhizoma could significantly inhibit the growth of 4 T1 breast cancer in mice(P< 0. 01),and exercise significantly promoted the anti-tumor activity of total alkaloids of Coptidis Rhizoma(P<0. 01),and reduced E2 and IL-1β levels in mice. Western blot and flow cytometry showed that the total alkaloids of Coptidis Rhizoma combined with exercise could down-regulate the protein expression levels of ERα,CDK4,CDK6,cyclin D1,CDK2 and cyclin E in cancer cells,block the transformation of G1/S in 4 T1 cell cycle,and inhibit DNA synthesis in breast cancer cells. The total alkaloids of Coptidis Rhizoma combined with exercise showed synergistic effect in inhibition of tumor growth in mice with orthotopically transplanted 4 T1 breast cancer.

Optimization of purification process for total alkaloids from Longzuan Tongbi Recipe with macroporous resin by central composite design and response surface method / 中草药

bjective To optimize the optimal purification technology for total alkaloids from Longzuan Tongbi Recipe (LTR). Methods Five types of macroporous resins were used to adsorb and desorb the total alkaloids in LTR, and the adsorption capacities, adsorption ratios and desorption ratios of total alkaloids were regarded as the indexes to screen out the suitable resin. The purification technology for total alkaloids was optimized from LTR by single factor investigation and central composite design and response surface method. Results The best purification technology of total alkaloid from LTR were determined as follows: the concentration of sample solution of 0.17 g/mL (crude drug) with pH 1.7, sample flow rate at 1 mL/min, washing impurity with 9-column volume of 80% ethanol at a flow rate of 1 mL/min. The mass scores of total alkaloids was 22.23% and the yield of that was 173.27 mg/g. Conclusion HPD100 resin has good purification effect on total alkaloids from LTR, which could provide reference for the further study of pharmacodynamics.

Pharmacokinetics study of brucine and strychnine in rat liver microsomes under different chemical environments / 中草药

Objective To study the enzyme kinetics of brucine (BRU) and strychnine (STR), total alkaloids, and monomers active components of BRU and STR in extracts of Strychnos nux-vomica in rat liver microsomes, and investigate metabolic differences between BRU and STR monomer, total alkaloids, and BRU and STR in extracts in rat liver microsomes. Methods The contents of BRU and STR in the monomers, total alkaloids, and extracts in the metabolic system in vitro were measured by LC-MS/MS method, and the kinetic parameters of the enzyme were calculated. Results Compared with monomer group, Km and Vmax value of BRU and STR in total alkaloids and extracts of S. nux-vomica were obviously decreased; Compared with total alkaloids, Km and Vmax value of BRU and STR monomers in S. nux-vomica were obviously increased. The CLint of BRU among total alkaloids and monomer extracts groups had no significantly difference; And the CLint of STR in three groups was successively decreased. Conclusion Total alkaloids, BRU and STR in S. nux-vomica extracts, and BRU and STR monomer can be metabolized by the liver microsomes, and the pharmacokinetical parameters of BRU and STR in all groups have significant differences in metabolism.

Study on preparation and evaluation in vitro of TASS-LLCN / 中草药

Objective To prepare a new kind of preparation formulation of the total alkaloids from seed of Strychni Semen (TASS), by which could improve their drug loading content and the transdermal function. Methods TASS-LLCN were prepared by hot solvent and high pressure homogeneous method. Using encapsulation efficiency (EE) measured by ultrafiltration as index, the prescription of TASS-LLCN was optimized by response surface methodology with central composite design, and the basic properties of the optimized TASS-LLCN was evaluated. The Franz diffuser method was used to compare the transdermal capacity of both LLCN gel and ordinary gel of TASS. Results The best prescription of TASS-LLCN was as follow: the content of glycerin monooleate (GMO) was 1 403.19 mg/mL, the ratio between GMO and F127 was 7.25:1, the drug loading content was 9.48% and the predicted encapsulation efficiency was 64.01%. The evaluation of basic properties of the optimized TASS-LLCN showed that the average grain diameter was about 186 nm, zeta potential was -33.1 mV, pH was 6.83, the stability was good. The transdermal experiment in vitro showed that the cumulate osmotic quantities in 24 h and the permeation rate of TASS-LLCN gel were both better than ordinary gel of TASS, which had obvious discrepancy (P < 0.05) and showed that LLCN could promote the absorption of active components; the skin retention volume of LLCN gel was still bigger than ordinary gel, which showed that LLCN gel of TASS could store in the skin and release the drug sustainedly. Conclusion TASS-LLCN could obviously enhance the drug loading content and the transdermal function. It is a potential new kind of preparation formulation which have the sustainable effect.

Study on the Purification Technology of Total Alkaloids from the Leaves of Nitraria sibirica with Macroporous Resin and Its Antioxidant Activity / 中国药房

OBJECTIVE:To optimize the purification technology of total alkaloids from the leaves of Nitraria sibirica with macroporous resin,and to evaluate its antioxidant activity. METHODS:Using the content of total alkaloids(by atropine sulfate)as index,static adsorption and analytical method were used to investigate the type of macroporous resin. Dynamic adsorption and analytical method were used to investigate the concentration of the sample solution,the pH of the sample solution,the flow rate of the sample solution,diameter-height ratio of column,sample capacity,the volume of elution water and volume fraction of elution solvent so as to optimize macroporous resin purification technology. Using vitamin C as positive control,scavenging capacity of total alkaloids to 1,1- diphenyl-2-diphenyl picryl hydrazinyl(DPPH)radicals was investigated to evaluate its antioxidant activity. RESULTS:The adsorption and desorption effects of HPD-450 macroporous resin on total alkaloids were the best. The optimal purification technology included the sample solution concentration 0.88 mg/mL,sample solution pH 6,sample solution flow rate 2 BV/h,diameter-height ratio of column 1∶8,sample capacity 5 BV,the volume of elution water 5 BV,elution solvent 30%ethanol. The content of total alkaloid was 16.94 mg/g,and IC50was (58.78 ± 3.00)μ g/mL by optimal purification technology. CONCLUSIONS:Optimized purification technology is stable and feasible,and can separate and purify total alkaloids from the leaves of N. sibirica. Purified total alkaloids display good antioxidant activities. The total alkaloids eluted with 30% ethanol show the strongest scavenging activity to DPPH radicals.

Inhibition Effect of Total Alkaloid, Total Phenolic Acid and Total Volatile Oil from Ligusticum chuanxiong on the Contraction of Isolated Uterine Smooth Muscle of Rats / 中国药房

OBJECTIVE: To investigate the inhibition effect of total alkaloid, total phenolic acid and total volatile oil from Ligusticum chuanxiong on the contraction of the isolated uterine smooth muscle of rats, and to provide reference for clarifying the promoting blood circulation to restore menstrual flow traditional efficacy of L.chuanxiong. METHODS: The isolated uterine smooth muscle of rats were collected and soaked in Locke's solution. Using dimethyl sulfoxide as blank control, verapamil hydrochloride as positive control, the contraction curves of the isolated uterine smooth muscle in rats were recorded with PowerLab electrophysiolograph. Inhibitory effects of total alkaloid, total phenolic acid (mass concentrations of 0. 025, 0. 05, 0. 1 mg/mL) and total volatile oil (0. 04, 0. 08, 0. 16 mg/mL) on the contraction of the isolated uterine smooth muscle of rats induced by oxytocin were evaluated by observing the changes of contraction activity and contraction tension. Inhibitory rates of contraction activity and contraction tension of uterine smooth muscle were calculated. RESULTS: The contraction activity and contraction tension of uterine smooth muscle in rats were increased significantly after treated with oxytocin (P<0. 05 or P<0. 01). Different concentrations of 3 types of compounds from L. chuanxiong all decreased contraction activity and contraction tension of uterine smooth muscle significantly (P<0. 05 or P<0. 01). Compared with blank control, except for 0. 025 mg/mL total phenolic acid, the inhibitory rates of contraction activity and contraction tension of uterine smooth muscle were all decreased significantly (P<0. 05 or P<0. 01)after treated with other concentration drugs or components. CONCLUSIONS: The total alkaloid, total phenolic acid and total volatile oil from L. chuanxiong show certain inhibitory effect on the contraction of isolated uterine smooth muscle in rats, which provide reference for promoting blood circulation to restore menstrual flow use of L. chuanxiong.

Optimization of Ultrasonic Extraction Process of Total Alkaloids from Oxytropis Falcata Bunge by Central Composite Design and Response Surface Method / 中国药师

Objective:To optimize the ultrasonic extraction process of total alkaloids from Oxytropis falcata bunge. Methods:The independent variables were solvents ratio, extracting time and ethanol concentration, and the dependent variable was content of total al-kaloids. Based on single factor tests, central composite design and response surface methodology was adopted to optimize the extraction technology. Results:The optimal extraction conditions were as follows: extracted twice with 36-fold amount of 72% ethanol ( contai-ning 1% acetic acid) at 60 ℃, and extracted 77 minutes each time. Under the above conditions, the content of total alkaloids was 2. 793 mg·g-1 with the bias ratio less than 2% when compared with the model predictions. Conclusion:Ultrasonic extraction process of total alkaloids from Oxytropis falcata Bunge optimized by central composite design and response surface method is simple, highly pre-cise, reliable and predictable.

Interactions of component (group) compatibility of Sanwu Huangqin Decoction in incubation model of rat liver microsomes / 中成药

AIM To study the interactions of component (group) compatibility of Sanwu Huangqin Decoction in incubation model of rat liver microsomes.METHODS With maackiain and ephedrine as internal standards,HPLC was adopted in the simultaneous content determination of total tlavonoids (baicalin,wogonoside,baicalein,wogonin and oroxylin A) from Scutellariae Radix and total alkaloids (oxymatrine,oxysophocarpine and matrine)from Sophorae flavescentis Radix in whole prescription component (group) compatibility and single medicinal material part (group) at seven time points (0,15,30,45,60,90 and 120 min),followed by the calculation of their in vitro metabolic rates.RESULTS In whole prescription component (group) compatibility,the contents of baicalin and wogonoside were first increased (0-0.5 h) and then decreased (0.5-2.0 h),those of baicalein,wogonin and oroxylin A showed increasing trends (more obvious within 0.5 h).The stable and gende metabolisms of various alkaloids reached balance within 20 min.CONCLUSION The bioavailability improvement and efficacy enhancement of total flavonoids from Scutellariae Radix may attribute to the compatibility with total alkaloids from Sophorae flavescentis Radix and total polysaccharides from Rehmanniae Radix in Sanwu Huangqin Decoction