Isolation, molecular characterization, immunological and anticoagulatant activities of polysaccharides from frankincense and its vinegar processed product.

Frankincense (FRA), the oily resin consisting of essential oils, boswellic acids (BAs) and polysaccharides, has been used to improve the blood circulation and relieve pain against carbuncles. According to the theory of traditional Chinese medicine, vinegar processed frankincense (VPF) can increase the effects of promoting blood circulation and relieving pain. Existing studies have carried out much on BAs and essential oils. However, the comparative analysis of polysaccharides from FRA and VPF has not been reported. In this paper, two polysaccharides were isolated and purified from FRA and the other two were from VPF, and their structures and physicochemical properties were analyzed. The immunological and anticoagulatant activities of the four polysaccharides were tested in RAW 264.7 cell and Sprague-Dawley rats, respectively. The polysaccharides purified from VPF showed better immunological and anticoagulatant activities than those in FRA. Therefore, polysaccharides may be one of the active substances for the synergistic effect of VPF.

Capsella bursa-pastoris (L.) Medic. extract alleviate cataract development by regulating the mitochondrial apoptotic pathway of the lens epithelial cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Capsella bursa-pastoris (L.) Medic. (CBP) is a cruciferous plant valuable in reducing fever, improving eyesight and calming the liver. This herb was recorded in the Compendium of Materia Medica for cataract treatment. AIM OF THE STUDY: To determine the effects and mechanism of CBP on cataract prevention and treatment using a selenite cataract model. MATERIALS AND METHODS: The main compounds in CBP extract were analyzed by UPLC, 1H-NMR and 13C-NMR spectroscopic techniques. Flavonoids formed a significant proportion of its compounds, thus necessitating an evaluation of their inhibitory effects on the development of cataract using a selenite cataract model. The protective effects of CBP flavonoids (CBPF) against oxidative damage and the modulation of mitochondrial apoptotic pathway were subsequently verified on H2O2-treated SRA01/04 lens epithelial cells. RESULTS: CBPF significantly alleviated the development of cataract by decreasing the MDA level and increasing the GSH-Px and SOD levels in the lens. It also inhibited H2O2-induced apoptosis in SRA01/04 cells, increased the expression of Bcl-2 protein and decreased the expressions of Caspase-3 and Bax proteins. CONCLUSION: CBPF exerts a significant preventive effect on cataract development by regulating the mitochondrial apoptotic pathway of the lens epithelial cells. It is thus a potent traditional Chinese medicine (TCM) whose application should be further developed for the clinical treatment of cataract.

Systematic characterization of the metabolites of defatted walnut powder extract in vivo and screening of the mechanisms against NAFLD by UPLC-Q-Exactive Orbitrap MS combined with network pharmacology.

ETHNOPHARMACOLOGICAL RELEVANCE: Walnut kernel, a well-known TCM, is often used after being defatted in tradition. And defatted walnut powder extract (DWPE) has the actions of tonifying the liver and kidney, dissipating stagnation and removing blood stasis, which has the effect on non-alcoholic fatty liver disease (NAFLD). However, the effective components of DWPE in vivo were unclear and the multiple mechanisms of DWPE against NAFLD have not been explored. AIM OF THE STUDY: The studies were performed to screen the effective substances in vivo by identification of the metabolites of DWPE in rats and to seek the potential mechanisms of DWPE on NAFLD by construction of the network pharmacology based on metabolites and verification of the highly correlated pathway. MATERIALS AND METHODS: To explore the effective substances in vivo, the metabolites of DWPE were identified in SD rats' bio-samples through UPLC-Q-Exactive Orbitrap MS. To analyze the mechanisms of DWPE on NAFLD, a Metabolite-Target-Disease network was established and the potential mechanisms were predicted. Then, highly correlated pathway was verified in animal and cells studies. RESULTS: A total of 52 metabolites of DWPE were identified in vivo, which were derived from gallic acid, ellagic acid (EA) and glansreginin A (Gla A). The possible metabolic pathways were phase Ⅰ (hydroxylation, hydrolyzation, etc) and phase Ⅱ metabolic reactions (methylation, sulfation and glucuronidation). Furthermore, in the network pharmacology, 54 core targets were enriched into pathways in cancer, nitrogen metabolism and other 9 pathways, which were essential pathways of DWPE against NAFLD. And the mechanism of nitrogen metabolism was verified in both of animal and cells studies. The results showed that DWPE could decline the concentration of ammonia and increase the expressions of carbonic anhydrase 2 (CA2) and carbamoylphosphate synthetase (CPS1) in nitrogen metabolism. CONCLUSION: Taken together, the study revealed the absorption components and their metabolic pathways and demonstrated the mechanism of nitrogen metabolism of DWPE on anti-NAFLD.

New tirucallane-type triterpenoids from the resin of Boswellia carteriiand their NO inhibitory activities.

Six new tirucallane-type triterpenoids (1-6), along with ten known triterpenoids, were isolated from methylene chloride extract of the resin of Boswellia carterii Birdw. By the application of the comprehensive spectroscopic data, the structures of the compounds were clarified. The experimental electronic circular dichroism spectra were compared with those calculated, which allowed to assign the absolute configurations. Compounds 5 and 6 possesed a 2, 3-seco tirucallane-type triterpenoid skeleton, which were first reported. Their inhibitory activity against NO formation in LPS-activated BV-2 cells were evaluated. Compound 9 showed appreciable inhibitory effect, with an IC50 value of 7.58 ± 0.87 µmol·L-1.

Anti-NAFLD effect of defatted walnut powder extract in high fat diet-induced C57BL/6 mice by modulating the gut microbiota.

ETHNOPHARMACOLOGICAL RELEVANCE: Walnut kernel has the actions of removing meteorism, dissipating stagnation and removing blood stasis and is used after being defatted in TCM. Defatted walnut powder extract (DWPE) has the abilities of anti-oxidation and lowering lipid levels in vivo. However, the effects and the potential mechanisms of DWPE on NAFLD have not been explored. AIM OF THE STUDY: The study were to investigate the anti-NAFLD effect of DWPE in high fat diet-induced C57BL/6 mice and demonstrate that whether DWPE developed the effect on anti-NAFLD by remodeling the compositions and abundances of gut microbiota. MATERIALS AND METHODS: The inhibitory effect of DWPE on the development of NAFLD was conducted on C57BL/6 mice with a high fat diet and the regulation effect of DWPE on gut microbiota was verified on pseudo-sterile mice with treatment of broad spectrum antibiotics. RESULTS: The results showed that the oral administration of DWPE remarkably alleviated hepatic lipid accumulation by decreasing the levels of TG, TC, LDL, MDA and increasing HDL. Meanwhile, the expressions of NF-κB and MAPKs family proteins were reduced by DWPE compared with HFD group. Otherwise, the efficacy of anti-NAFLD of DWPE was significantly decreased after treatment of antibiotics, which indicated the key role of gut microbiota in the therapeutic process. Furthermore, sequencing of 16S rRNA gene revealed that DWPE could revert the decreased relative abundance of gut microbiota caused by the long term of a high fat diet. And the disordered microflora was remodeled by DWPE including the reduction of Erysipelotrichia, Firmicutes and Actinobacteria as well as the increment of Bacteroidetes, Clostridiales, Bacteroidales S24-7, Prevotellaceae and Bacteroides. CONCLUSION: Taken together, DWPE had a preventing effect on NAFLD, which might be associated with the regulation of gut microbiota.

Morning glory seed keeps laxative effect while retains less subchronic toxicity after being fried.

ETHNOPHARMACOLOGICAL RELEVANCE: Morning glory seed (MGS), has been widely used in treating constipation especially towards children. Clinically, people usually take fried MGS (MGSF) in formulas to reduce its side effect. However, the safety of MGSF other than MGS has yet to be explored. OBJECTIVE: The study aimed to reveal the potential mechanisms of using MGSF instead of MGS basing on chemistry, pharmacodynamics and toxicology. METHODS: The chemical compositions of the extracts of MGS and MGSF were compared using UPLC-Q-TOF/MS method. Simultaneously, to prove the availability and safety of MGSF, we investigated the laxative effect and subchronic toxicity of MGS and MGSF and addressed the mechanism of laxative effect of them. RESULTS: In this study, less phenolic acids and more fatty acids were detected in MGSF compared with the compounds in MGS. Moreover, we found that MGS group had stronger laxative effect than MGSF group via downregulating the expression of AQP3 protein. As for subchronic toxicity test, the body weights of MGS group were lower than MGSF group. In serum biochemistry and histopathological examinations, MGS group could cause more serious toxicity in liver, kidney and colon than MGSF group with higher values of BUN, Cr, AST and ALP. CONCLUSION: Based on the findings in this study, MGSF with varied compounds contents could still keep the laxative effect while retain less subchronic toxicity, which emphasized the necessity of processing and provided an insight into the rational use of MGSF in clinical practice.

Systematically characterize the substance basis of Jinzhen oral liquid and their pharmacological mechanism using UPLC-Q-TOF/MS combined with network pharmacology analysis.

Jinzhen oral liquid (JZ) is a classical traditional Chinese medicine formula used for the treatment of children lung disease. However, the effective substance of JZ is still unclear. In this study, we used lung injury rat model to study the protective effect of JZ, through UPLC-Q-TOF/MS detection coupled with metabolic research and network pharmacology analysis. Fortunately, 31 absorbed prototype constituents and 41 metabolites were identified or tentatively characterized based on UPLC-Q-TOF/MS analysis, and the possible metabolic pathways were hydroxylation, sulfation and glucuronidation. We optimized the data screening in the early stage of network pharmacology by collecting targets based on adsorbed constituents, and further analyzed the main biological processes and pathways. 24 selected core targets were frequently involved in reactive oxygen species metabolic process, dopaminergic synapse pathway and so on, which might play important roles in the mechanisms of JZ for the treatment of lung injury. Overall, the absorbed constituents and their possible metabolic pathways, as well as the absorbed constituent-target-disease network provided insights into the mechanisms of JZ for the treatment of lung injury. Further studies are needed to validate the biological processes and effect pathways of JZ.

Xanthones from Garcinia paucinervis with in vitro anti-proliferative activity against HL-60 cells.

Three new xanthones, paucinervins H-J (1-3), as well as eleven known compounds (4-14), were isolated from the leaves of Garcinia paucinervis. The structures of the new compounds (1-3) were elucidated by 1D, 2D NMR spectra and HR ESIMS. In vitro antiproliferative activity against human promyelocytic leukemia HL-60 cells was tested, among which, compounds 2, 5, 6 and 7 exhibited strong growth inhibitory effects with GI50 values ranging from 1.30 to 9.08 µM, respectively. Preliminary SARs were also discussed.

Comparative studies of pharmacokinetics and anticoagulatory effect in rats after oral administration of Frankincense and its processed products.

ETHNOPHARMACOLOGICAL RELEVANCE: Frankincense (FRA), Ruxiang, is the resin of Boswellia carterii Birdw and Boswellia bhaw-dajiana Birdw which has been used for centuries as formulas to improve the circulation and to relieve pain against carbuncles. Stir-fried Frankincense (SFF) and vinegar processed Frankincense (VPF) are two major processed Frankincense, and the processing procedures reportedly enhance the curative efficacy or reduce the side effects of FRA. This paper describes the comparisons in plasma pharmacokinetic behaviors of 11-keto-ß-boswellic acid (KBA) and 3-acetyl-11-keto-ß-boswellic acid (AKBA) in FRA and its processed products, and their effects on coagulation factors and blood clotting tetrachoric, using an acute cold blood-stasis animal model after oral administration of FRA, SFF, and VPF. MATERIALS AND METHODS: For pharmacokinetic study, Sprague-Dawley (SD) rats were randomly divided into three groups, including group FRA, group SFF and group VPF. And the plasma samples were analyzed by HPLC. For study of anticoagulatory effect, SD rats were randomly divided into six groups, including control, acute cold blood-stasis model, Fu-fang-dan-shen tablet- (0.75g/kg), FRA-, SFF-, and VPF-treated (2.7g/kg) groups, respectively. The serum contents of thrombin-antithrombin complex (TAT), D-dimer (D-D), and prostacyclin (PGI2) of each group were measured by ELISA. The values of prothrombin time (PT), thrombin time (TT), activated partial thromboplastin time (APTT) and fibrinogen (FIB) were also assessed by hematology analyzer. RESULTS: Significantly increased levels of Cmax, AUC, T1/2, and MRT were found in rats treated with the processed products. In addition, decreased levels of D-D and TAT and increased contents of PGI2 were observed in rats given FRA and its processed products, compared with that of the model group. Moreover, VPF improved anticoagulation more than SFF in the animals. CONCLUSIONS: The observed improvement of anticoagulation by processed FRA may result from the increased absorption and bioavailability of triterpenoids.

Xiaochaihu Decoction attenuates the vicious circle between the oxidative stress and the ALP inactivation through LPS-catecholamines interactions in gut, liver and brain during CCI4+ethanol-induced mouse HCC.

BACKGROUND: Xiaochaihu Decoction (XCHD) prevents hepatocarcinogenesis in association with inhibition of oxidative stress. However, alkaline phosphatase (ALP) activity, lipopolysaccharides (LPS)-catecholamines (CA) interactions in gut, liver and brain may play an important role in the status of oxidative stress. This study was to assess whether XCHD attenuates the vicious circle between oxidative stress and ALP inactivation through LPS-CA interactions. METHODS: Hepatocellular carcinoma group (HCC) were induced by CCI4 + ethanol; HCC with Liver Depression and Spleen Deficiency (HCC + LDSD) were induced by squeezing tails (30 min/day), solitary breeding and intermittent fasting on the basis of HCC; XCHD was administered after 4 weeks of the HCC + LDSD. The degree of tissue injury were studied using a scoring system, and brain weights were measured. Peroxynitrite (ONOO(-)), malondialdehyde (MDA), 4-hydroxy-3-methoxymandelic acid (VMA, CA metabolites), lipopolysaccharide-phosphate (LPS-P), ALP activity (ALP-A) and Concanavalin A (ConA)-binding rate of ALP (ALP-C) were determined by colorimetric method and lectin (ConA) affinity precipitation method. RESULTS: More injuries and ONOO(-), MDA, VMA, LPS-P, ALP-C were increased, ALP-A were decreased in the gut, liver and brain of HCC group, the most in HCC + LDSD group, after treatment with XCHD, all of which were improved. A positive association found between gut-liver-brain injury and ONOO(-), MDA, VMA, LPS-P, ALP-C, between ONOO(-), MDA, VMA, LPS-P and ALP-C in the gut, liver and brain, and a negative association found between gut-liver-brain injury and ALP-A, between ALP-A and ONOO(-), MDA, VMA, LPS-P, ALP-C in the gut, liver and brain. CONCLUSIONS: XCHD can attenuates the vicious circle between the oxidative stress, nitrosative stress, N-glycan deficiency and inactivation of ALP through LPS-CA interactions in gut, liver and brain.

Four new cytotoxic tetrahydrofuranoid lignans from Sinopodophyllum emodi.

Four new tetrahydrofuranoid lignans, (-)-tanegool-7'-methyl ether ( 1), (+)-7'-methoxylariciresinol (2), sinolignan C (3), and epipinoresinol-4,4'-di- O- ß- D-glucopyranoside (4), were isolated from the roots and rhizomes of SINOPODOPHYLLUM EMODI together with one known lignan (5). Their structures and stereochemistry were elucidated on the basis of spectroscopic and mass spectrometric evidence. The isolation of compounds 1- 5 represents the first report of tetrahydrofuran lignans from the genus SINOPODOPHYLLUM. The cytotoxic activities of all isolated compounds were evaluated against HeLa and KB cell lines, and compound 1 showed the most potent cytotoxicity with IC50 values of 9.7 µM and 4.7 µM, respectively.

Three new phenolic compounds from the lichen Thamnolia vermicularis and their antiproliferative effects in prostate cancer cells.

Three new phenolic compounds, thamnoliadepsides A (1), B (2), and thamnolic acid A (3), and seven known compounds, everninic acid (4), baeomycesic acid (5), ß-orcinol (6), ß-resorcylic acid (7), ethyl orsellinate (8), squamatic acid (9), and vermicularin (10), were isolated from the lichen Thamnolia vermicularis (Sw.) Ach. ex Schaerer. Their structures were determined based on spectroscopic analysis, including 2D-NMR experiments and HR-MS techniques. Compound 1 inhibited growth of prostate cancer cells and bonded to G-quadruplex DNA based on NMR determination.

Triterpenes and xanthones from the stem bark of Garcinia tetralata.

A new compound, 3ß,18,19ß-trihydroxylupane, was isolated from Garcinia tetralata, along with five known compounds, garcinexanthone B, morolic acid acetate, toxyloxanthone A, 6,11-dihydroxy-2,2-dimethylpyrano[3,2-c]xanthen-7(2H)-one, and 1,4-dihydroxy-5,6-dimethoxy-xanthone. The structure of the new compound was established by extensive spectroscopic techniques.

Effect of Bu-Zhong-Yi-Qi-Tang on deficiency of N-glycan/nitric oxide and islet damage induced by streptozotocin in diabetic rats.

AIM: To investigate the effect of Bu-Zhong-Yi-Qi-Tang (Decoction for Reinforcing Middle Jiao and Replenishing Qi) on deficiency of N-glycan/nitric oxide (NO) and islet damage induced by injecting two medium doses of streptozotocin (STZ). METHODS: Diabetes was induced by intraperitoneal injection of STZ at 55 mg/kg on day 1 and day 8. Islet damage was evaluated using a scoring system. Nitrite, nitrate, alpha-mannosidase and amylase activities were measured by colorimetry. N-glycan patterns of amylase were determined with lectin [ConA, pisum sativum agglutinin (PSA), peanut agglutinin (PNA), and lens culinaris agglutinin (LCA)] affinity precipitation method. RESULTS: Severe islet necrosis and mild islet atrophy were observed in diabetic rats. The number and size of islets, the activities of alpha-mannosidase, amylase and nitrite were decreased, while the binding of PNA and LCA to amylase was increased. All of which were improved after treatment with Bu-Zhong-Yi-Qi-Tang. Islet damage was significantly correlated with nitrite, nitrate, alpha-mannosidase, amylase and the binding of LCA, PNA, and PSA to amylase. CONCLUSION: STZ-induced islet damage is related to N-glycan deficiency in proteins by blocking alpha-mannosidase activity and no deficiency, accumulation of unfolded proteins, and endoplasmic reticulum stress and activation of cellular signals, all of which are improved after treatment with Bu-Zhong-Yi-Qi-Tang.

[Current quality research situation of precious Chinese patent medicine].

Along with the development of modern medicinal manufacture and pharmaceutical analysis technology, the precious Chinese patent medicine is also growing quickly. However, after the investigation of the dosage-forms and relative government standards, we find that the precious Chinese patent medicine needs more variety in administration and more precise and full-scale analysis standards based on the chemical materials because of the contradiction between the "global thoughts" logic in traditional Chinese medicine (TCM) theory and the "local study" of modern pharmaceutical analysis technology. Currently rising theories, metabonomics and coordination chemistry of TCM, with the superiority of entirety, can not only fulfill the requirement for " local study" but also accord with the "global thoughts" logic in TCM theory. They will be the reasonable guide for the development of the precious Chinese patent medicine.

Simultaneous qualitative and quantitative analysis of commercial bistorta rhizome and its differentiation from closely related herbs using TLC and HPLC-DAD fingerprinting.

A qualitative and quantitative analysis method was established to improve quality assessment standards for Rhizoma Polygoni Bistortae (Polygonum bistorta L.) and differentiate commercial bistorta rhizome from closely related herbs by TLC and HPLC-DAD fingerprinting. Three compounds including phenolic acid and flavane were identified by comparison with standard compounds and quantified simultaneously by HPLC-DAD simultaneously. A comprehensive validation of the method that included sensitivity, linearity, repeatability and recovery was conducted. Paris polyphylla SM., a herb often mixed with Polygonum bistorta L. in China due to their same popular name "Caoheche" in history, was successfully distinguished by thin-layer chromatography (TLC) fingerprinting of the petroleum-soluble fraction. Polygonum paleaceum WALL., another herb often mixed with Polygonum bistorta L. due to their similar external appearances, was distinguished by HPLC fingerprinting.

[Chemical constituents from barks of Garcinia tetralata].

OBJECTIVE: To study the chemical constituents of the barks of Garcinia tetralata. METHOD: Compounds were isolated and purified repeatedly by silica gel and ODS column chromatography and their chemical structures were elucidated by their physicochemical properties and spectral data analysis. RESULT: Ten compounds were obtained and identified as 1, 7-dihydroxyxanthone (1), buchanaxanthone (2), garciniaxanthone H (3), 1, 2, 5-trihydroxyxanthone (4), subelliptenone H (5), 6-desoxyjacareubin (6), 6-deoxyisojacreubin (7), subelliptenone G (8), methyl orsellinate (9) and beta-sitosterol (10). CONCLUSION: Compounds 1-10 were isolated from this plant and compound 9 was obtained from the genus Garcinia for the first time.

[Studies on chemical constituents from herbs of Botrychium lanuginosum].

OBJECTIVE: To study the chemical constituents of Botrychium lanuginosum. METHOD: Various chromatographic techniques were used to isolate and purify the constituents. The structures were elucidated by chemical evidence and spectroscopic methods. RESULT: Ten compounds were isolated from the 95% ethanol extract of the herb of B. lanuginosum and their structures were elucidated as 30-nor-21beta-hopan-22-one (1), beta-sitosterol (2), luteolin (3), thunberginol A (4), apigenin (5), (6'-O-palmitoyl)-sitosterol-3-O-beta-D-glucoside (6), daucosterol (7), 1-O-beta-D-glucopyranosyl-(2S, 3R, 4E, 8Z)-2-[(2R-hydroxy hexadecanoyl) amino]-4, 8-octadecadiene-1, 3-diol (8), luteolin-7-O-glucoside (9), sucrose (10). CONCLUSION: Compounds 1-10 were isolated from this genus for the first time.