Oligomeric proanthocyanidins ameliorates osteoclastogenesis through reducing OPG/RANKL ratio in chicken's embryos.

Skeletal disorders can seriously threaten the health and the performance of poultry, such as tibial dyschondroplasia (TD) and osteoporosis (OP). Oligomeric proanthocyanidins (OPC) are naturally occurring polyphenolic flavonoid compounds that can be used as potential substances to improve the bone health and the growth performance of poultry. Eighty 7-day-old green-eggshell yellow feather layer chickens were randomly divided into 4 groups: basal diet and basal diet supplementation with 25, 50, and 100 mg/kg OPC. The results have indicated that the growth performance and bone parameters of chickens were significantly improved supplementation with OPC in vivo, including the bone volume (BV), the bone mineral density (BMD) and the activities of antioxidative enzymes, but ratio of osteoprotegerin (OPG)/receptor activator of NF-κB (RANK) ligand (RANKL) was decreased. Furthermore, primary bone marrow mesenchymal stem cells (BMSCs) and bone marrow monocytes/macrophages (BMMs) were successfully isolated from femur and tibia of chickens, and co-cultured to differentiate into osteoclasts in vitro. The osteogenic differentiation derived from BMSCs was promoted treatment with high concentrations of OPC (10, 20, and 40 µmol/L) groups in vitro, but emerging the inhibition of osteoclastogenesis by increasing the ratio of OPG/RANKL. In contrary, the osteogenic differentiation was also promoted treatment with low concentrations of OPC (2.5, 5, and 10 µmol/L) groups, but osteoclastogenesis was enhanced by decreasing the ratio of OPG/RANKL in vitro. In addition, OPG inhibits the differentiation and activity of osteoclasts by increasing the autophagy in vitro. Dietary supplementation of OPC can improve the growth performance of bone and alter the balance of osteoblasts and osteoclasts, thereby improving the bone health of chickens.

Baicalin and N-acetylcysteine regulate choline metabolism via TFAM to attenuate cadmium-induced liver fibrosis.

(Background): Cadmium is an environmental pollutant associated with several liver diseases. Baicalin and N-Acetylcysteine have antioxidant and hepatoprotective effects. (Purpose): However, it is unclear whether baicalin and N-Acetylcysteine can alleviate Cadmium -induced liver fibrosis by regulating metabolism, or whether they exert a synergistic effect. (Study design): We treated Cadmium-poisoned mice with baicalin, N-Acetylcysteine, or baicalin+ N-Acetylcysteine. We studied the effects of baicalin and N-Acetylcysteine on Cadmium-induced liver fibers and their specific mechanisms. (Methods): We used C57BL/6 J mice, and AML12, and HSC-6T cells to establish in vitro assays and in vivo models. (Results): Metabolomics was used to detect the effect of baicalin and N-Acetylcysteine on liver metabolism, which showed that compared with the control group, the Cadmium group had increased fatty acid and amino acid levels, with significantly reduced choline and acetylcholine contents. Baicalin and N-Acetylcysteine alleviated these Cadmium-induced metabolic changes. We further showed that choline alleviated Cadmium -induced liver inflammation and fibrosis. In addition, cadmium significantly promoted extracellular leakage of lactic acid, while choline alleviated the cadmium -induced destruction of the cell membrane structure and lactic acid leakage. Western blotting showed that cadmium significantly reduced mitochondrial transcription factor A (TFAM) and Choline Kinase α(CHKα2) levels, and baicalin and N-Acetylcysteine reversed this effect. Overexpression of Tfam in mouse liver and AML12 cells increased the expression of CHKα2 and the choline content, alleviating and cadmium-induced lactic acid leakage, liver inflammation, and fibrosis. (Conclusion): Overall, baicalin and N-Acetylcysteine alleviated cadmium-induced liver damage, inflammation, and fibrosis to a greater extent than either drug alone. TFAM represents a target for baicalin and N-Acetylcysteine, and alleviated cadmium-induced liver inflammation and fibrosis by regulating hepatic choline metabolism.

Optimization of Ultrasonic-Assisted Extraction of Chlorogenic Acid from Tobacco Waste.

Using tobacco waste as raw material, the ultrasonic-assisted extraction of chlorogenic acid was optimized by response surface methodology (RSM). After repeated freezing and thawing of tobacco waste twice, the effect of pH value, ethanol volume fraction, temperature and extraction time on the extraction rate of chlorogenic acid was investigated by a single factor experiment. On the basis of this, the factors affecting the yield of chlorogenic acid were further optimized by using RSM. The optimum extraction conditions for chlorogenic acid were set at pH = 4.1, ethanol volume fraction was 49.57% and extraction time was 2.06 h. Under the above conditions, the extraction rate of chlorogenic acid could reach 0.502%, which was higher than traditional extraction and unpretreated ultrasonic extraction. All these results can be used as a reference for the extraction of effective ingredients in tobacco waste.

Ligustilide attenuates nitric oxide-induced apoptosis in rat chondrocytes and cartilage degradation via inhibiting JNK and p38 MAPK pathways.

Ligustilide (LIG) is the main lipophilic component of the Umbelliferae family of pharmaceutical plants, including Radix angelicae sinensis and Ligusticum chuanxiong. LIG shows various pharmacological properties associated with anti-inflammation and anti-apoptosis in several kinds of cell lines. However, the therapeutic effects of LIG on chondrocyte apoptosis remain unknown. In this study, we investigated whether LIG had an anti-apoptotic activity in sodium nitroprusside (SNP)-stimulated chondrocyte apoptosis and could delay cartilage degeneration in a surgically induced rat OA model, and elucidated the potential mechanisms. In vitro studies revealed that LIG significantly suppressed chondrocyte apoptosis and cytoskeletal remodelling, which maintained the nuclear morphology and increased the mitochondrial membrane potential. In terms of SNP, LIG treatment considerably reduced the expression levels of cleaved caspase-3, Bax and inducible nitric oxide synthase and increased the expression level of Bcl-2 in a dose-dependent manner. The LIG-treated groups presented a significantly suppressed expression of activating transcription factor 2 and phosphorylation of Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MAPK). The inhibitory effect of LIG was enhanced by the p38 MAPK inhibitor SB203580 or the JNK inhibitor SP600125 and offset by the agonist anisomycin. In vivo studies demonstrated that LIG attenuated osteoarthritic cartilage destruction by inhibiting the cartilage chondrocyte apoptosis and suppressing the phosphorylation levels of activating transcription factor 2, JNK and p38 MAPK. This result was confirmed by histological analyses, micro-CT, TUNEL assay and immunohistochemical analyses. Collectively, our studies indicated that LIG protected chondrocytes against SNP-induced apoptosis and delayed articular cartilage degeneration by suppressing JNK and p38 MAPK pathways.

A myrsinol diterpene isolated from a traditional herbal medicine, LANGDU reverses multidrug resistance in breast cancer cells.

ETHNOPHARMACOLOGICAL RELEVANCE: LANGDU, a Chinese traditional herbal medicine, was the dried roots of Euphorbia prolifera Buch-Ham. The herbal medicine has been used as anti-cancer and anti-inflammatory drug in local folk medicine for several hundred years. AIM OF THE STUDY: P-glycoprotein (P-gp) is a transmembrane exporter, which can expel a variety of anti-cancer drugs. Over-expressed P-glycoprotein in cancer cells impairs the effect of cancer chemotherapy and results in multidrug resistance (MDR). To elucidate the effect of LANGDU on MDR cancer cells, the constituents of Euphorbia prolifera Buch-Ham were analyzed. We found that a myrsinol diterpene, J196-10-1 could reverse multidrug resistance. MATERIALS AND METHODS: Cytotoxicity assays were performed to measure reversal efficiency of J196-10-1. Efflux assay, ATPase assay, and real-time PCR were used to elucidate the mechanism of the chemical. RESULTS: J196-10-1 could reverse the resistance to daunorubicin, vincristine, and topotecan effectively. The diterpene inhibited P-gp mediated efflux and did not alter transcription of the target gene significantly. The compound stimulated ATP hydrolysis at a low concentration and inhibited it at a high concentration. CONCLUSIONS: J196-10-1 inhibits P-gp competitively and reverses P-gp induced MDR in breast cancer cells.

A myrsinol diterpene isolated from Euphorbia prolifera reverses multidrug resistance in breast cancer cells.

P-glycoprotein (P-gp), a member of ATP-Binding Cassette transporter superfamily, can expel a variety of anti-cancer drugs so that it impairs the effect of cancer chemotherapy and results in multidrug resistance (MDR). The P-gp inhibitors are important to circumvent MDR and improve efficacy of cancer chemotherapy. The dried root of Euphorbia prolifera Buch.-Ham. has been used to treat cancer and inflammation in Chinese folk medicine for several hundred years. A myrsinol diterpene derived from Euphorbia prolifera Buch.-Ham. (J196-9-4) could modulate multidrug resistance. Cytotoxicity assays were performed to measure the reversal efficiency of J196-9-4. Efflux assay and ATPase assay were used to elucidate the mechanism of the chemical. J196-9-4 potentiated cytotoxicity of anti-cancer drugs in the P-gp over-expressing resistant breast cancer cell line MCF-7/Adr as compared to MCF-7 cells. Concentrations of 5 and 10 µM J196-9-4 could reverse the resistance to daunorubicin, vincristine, and topotecan significantly. Since J196-9-4 inhibited P-gp mediated efflux and stimulated ATP hydrolysis, J196-9-4 was a substrate of P-gp. Thus J196-9-4 is a competitive inhibitor of P-gp and reverses multidrug resistance induced by the transporter.

An isoquinoline alkaloid from the Chinese herbal plant Corydalis yanhusuo W.T. Wang inhibits P-glycoprotein and multidrug resistance-associate protein 1.

Overexpression of P-glycoprotein (P-gp) and multidrug resistance-associate protein 1 (MRP1) is a major mechanism leading to multidrug resistance (MDR) of cancer cells. These transporters expel anti-cancer drugs and greatly impair therapeutic efficacy of chemotherapy. A Chinese herbal plant Yanhusuo (Corydalis yanhusuo W.T. Wang, YHS) is frequently used in functional food and traditional Chinese medicine to improve the efficacy of chemotherapy. The objective of this work was to study effects of glaucine, an alkaloid component of YHS, on P-gp and MRP1 in resistant cancer cells. The resistant cancer cell line, MCF-7/ADR and corresponding parental sensitive cells were employed to determine reversal properties of glaucine. Glaucine inhibits P-gp and MRP1-mediated efflux and activates ATPase activities of the transporters, indicating that it is a substrate and inhibits P-gp and MRP1 competitively. Furthermore, glaucine suppresses expression of ABC transporter genes. It reverses the resistance of MCF-7/ADR to adriamycin and mitoxantrone effectively.

DHA derivatives of fish oil as dietary supplements: a nutrition-based drug discovery approach for therapies to prevent metabolic cardiotoxicity.

INTRODUCTION: During the early 1970s, Danish physicians Jorn Dyerberg and colleagues observed that Greenland Eskimos consuming fatty fishes exhibited low incidences of heart disease. Fish oil is now one of the most commonly consumed dietary supplements. In 2004, concentrated fish oil was approved as a drug by the FDA for the treatment of hyperlipidemia. Fish oil contains two major omega-3 fatty acids: eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). With advancements in lipid concentration and purification techniques, EPA- or DHA-enriched products are now commercially available, and the availability of these components in isolation allows their individual effects to be examined. Newly synthesized derivatives and endogenously discovered metabolites of DHA exhibit therapeutic utility for obesity, metabolic syndrome and cardiovascular disease. AREAS COVERED: This review summarizes our current knowledge on the distinct effects of EPA and DHA to prevent metabolic syndrome and reduce cardiotoxicity risk. Since EPA is an integral component of fish oil, we will briefly review EPA effects, but our main theme will be to summarize effects of the DHA derivatives that are available today. We focus on using nutrition-based drug discovery to explore the potential of DHA derivatives for the treatment of obesity, metabolic syndrome and cardiovascular diseases. EXPERT OPINION: The safety and efficacy evaluation of DHA derivatives will provide novel biomolecules for the drug discovery arsenal. Novel nutritional-based drug discoveries of DHA derivatives or metabolites may provide realistic and alternative strategies for the treatment of metabolic and cardiovascular disease.

Four new sesquiterpenes from Commiphora myrrha and their neuroprotective effects.

Four new sesquiterpenes, myrrhterpenoids K-N (1-4) have been isolated from the resin of Commiphora myrrha. Their structures and relative configurations were elucidated by spectroscopic methods (IR, ESIMS, HR-ESIMS, 1D and 2D NMR). Compounds 1 and 4 showed neuroprotective effects against MPP(+)-induced neuronal cell death in SH-SY5Y cells.

Iridoids from the roots of Valeriana jatamansi and their neuroprotective effects.

Three new iridoids, valeriandoids A-C (1-3), together with three known analogues (4-6), were isolated from the roots of Valeriana jatamansi. Their structures and relative configurations were elucidated by spectroscopic methods (IR, ESIMS, HRESIMS, 1D and 2D NMR) and by comparison of their NMR data with those of related compounds. All the isolated compounds were evaluated for their neuroprotective effects and compounds 1, 3, 4 and 6 showed moderate neuroprotective effects.

Sesquiterpenoids from the resinous exudates of Commiphora myrrha and their neuroprotective effects.

Ten new furanosesquiterpenoids, myrrhterpenoids A-J (1- 10), together with eight known analogues (11- 18), were isolated from the resinous exudates of COMMIPHORA MYRRHA. Their structures and relative configurations were elucidated by spectroscopic methods (IR, ESIMS, HRESIMS, 1D and 2D NMR) and by the ChemDraw 3D modeling using MM2. All isolated furanosesquiterpenes showed neuroprotective effects against MPP (+)-induced neuronal cell death in SH-SY5Y cells.

Piperine, a piperidine alkaloid from Piper nigrum re-sensitizes P-gp, MRP1 and BCRP dependent multidrug resistant cancer cells.

Over-expression of P-gp, MRP1 and BCRP in tumor cells is one of the important mechanisms leading to multidrug resistance (MDR), which impairs the efficacy of chemotherapy. P-gp, MRP1 and BCRP are ABC (ATP-Binding Cassette) transporters, which can expel a variety of lipophilic anti-cancer drugs and protect tumor cells. During a screening of MDR reversal agents among alkaloids of various structural types, a piperidine alkaloid, piperine (a main piperidine alkaloid in Piper nigurm) was identified as an inhibitor. Piperine can potentiate the cytotoxicity of anti-cancer drugs in resistant sublines, such as MCF-7/DOX and A-549/DDP, which were derived from MCF-7 and A-549 cell lines. At a concentration of 50 µM piperine could reverse the resistance to doxorubicin 32.16 and 14.14 folds, respectively. It also re-sensitized cells to mitoxantrone 6.98 folds. In addition, long-term treatment of cells by piperine inhibits transcription of the corresponding ABC transporter genes. These results suggest that piperine can reverse MDR by multiple mechanisms and it may be a promising lead compound for future studies.

Neuroprotective labdane diterpenes from Fritillaria ebeiensis.

By bioactivity-guided isolation, two labdane diterpenes (1 and 2) have been isolated from the bulbs of Fritillaria ebeiensis. Their structures were elucidated as 6α,7ß-dihydroxy-labda-8(17),12(E),14-triene (1) and 6-oxo-2α-hydroxy-labda-7,12(E), 14-triene (2), on the basis of spectroscopic data analysis (IR, ESI-MS, HR-ESI-MS, 1D and 2D NMR). Both of the isolates showed neuroprotective effects against MPP(+)-induced neuronal cell death in human dopaminergic neuroblastoma SH-SY5Y cells.

Sesquiterpenes from Vladimiria souliei and their inhibitory effects on NO production.

As a part of our ongoing search for plant-derived compounds that inhibit nitric oxide production, the methanol extract of the roots of Vladimiria souliei was found to show significant inhibitory effects on INF-γ-induced nitric oxide production in murine macrophage RAW264.7 cells. Bioactivity-guided isolation of the extract yielded two most active sesquiterpenes, including a new compound, named souliene A (1) and alismol (2). Their structures were elucidated by spectroscopic methods (IR, ESIMS, HRESIMS, 1D and 2D NMR). Two isolates showed promising inhibitory effects on INF-γ-induced nitric oxide production in murine macrophage RAW264.7 cells.

The beta-carboline alkaloid harmine inhibits BCRP and can reverse resistance to the anticancer drugs mitoxantrone and camptothecin in breast cancer cells.

Multidrug resistance (MDR), mediated by highly expressed ABC transporters, is one of the most important mechanisms in tumor cells. Breast cancer resistance protein (BCRP) is a member of the ABC transporter family. This transporter expels different kinds of lipophilic anticancer drugs, which have diffused into the cells. In this study, 96-well plate based assays and flow cytometry analysis were employed to screen natural products for BCRP inhibition. The beta-carboline alkaloid harmine inhibited BCRP in a BCRP overexpressing breast cancer cell line (MDA-MB-231). Harmine reduced resistance to the anticancer drugs mitoxantrone and camptothecin mediated by BCRP and might be an interesting new reversal agent. Harmine did not inhibit P-glycoprotein (P-gp) mediated drug efflux.

[Study toxicity-attenuating effect and dose-toxicity relationship of rhubarb by processing based on correspondence analysis].

OBJECTIVE: To study the toxicity-attenuating effect and the dose-toxicity relationship of rhubarb by processing based on correspondence. METHOD: The effects of different processed materials of rhubarb on the hepatic and renal functions of mice was researched in a way of parallel comparison, as well as the chemical alteration induced by processing was observed. Correspondence analysis, a kind of multivariate statistical analysis, was performed to explore the dose-toxicity relationship of processed materials of rhubarb. RESULT: No obvious toxic effect was found in mice after single intragastric administration of crude drug of rhubarb at dosage of 76 g x kg(-1), while some lesions to liver and kidney tissues were observed in mice after repeated administration of rhubarb and its processed materials for 14 days at the same dosage. In the correspondence analysis diagram, it could be deduced that there was definite dose-toxicity relationship of processed rhubarb as the distance between trial groups and control group increased along with the dosage and the toxicity. The distance of the processed rhubarb showed as the following consequence: crude drug of rhubarb > vinegar-processed rhubarb > alcohol-processed rhubarb > steamed rhubarb > carbonized rhubarb > Qingning pian. Theerefoer, the toxicity of processed rhubarb was much lower than that of crude drug and the extent of toxicity attenuation was related to the processing intensity. Meanwhile, the toxicity-attenuating effect of processed rhubarb was related to the decline of the contents of both anthraquinone glycosides and tannins, and the former was contributed remarkably to toxicity. CONCLUSION: The toxicity-attenuating effect of processed rhubarb was verified in this study and the toxicity of steamed rhubarb attenuated notably while the pharmacological substances degraded little. The correspondence analysis would be useful to assess the pharmacological and toxic effects with multiple indexes of traditional Chinese medicines.

Lobeline, a piperidine alkaloid from Lobelia can reverse P-gp dependent multidrug resistance in tumor cells.

Multidrug resistance (MDR) can limit efficacy of chemotherapy. The best studied mechanism involves P-gp (P-glycoprotein) mediated drug efflux. This study focuses on MDR reversal agents from medicinal plants, which can interfere with P-gp. Rhodamine 123 accumulation assay and flow cytometry analysis were employed to screen for P-gp dependant efflux inhibitors. Lobeline, a piperidine alkaloid from Lobelia inflata and several other Lobelia species, inhibited P-gp activity. MDR reversal potential of lobeline could be demonstrated in cells treated with doxorubicin in that lobeline can sensitize resistant tumor cells at non-toxic concentrations. However, lobeline cannot block BCRP (Breast Cancer Resistance Protein) dependent mitoxantrone efflux. Lobeline could be a good candidate for the development of new MDR reversal agents.

[Fingerprints of different samples of processed Rhizoma Coptidi by HPLC].

OBJECTIVE: To set up the HPLC fingerprints of different samples of processed Rhizoma Coptidi (RC) in order to offer evidence for identifying different processed RC and preliminary discussion on processing mechanism. METHOD: Seven different samples of processed RC were determined by HPLC and the results were analyzed by Hierarchical clustering and similarity evaluation system for chromatographic fingerprint of TCM. RESULT: The characteristic of common peaks for fingerprints of different samples of processed RC is distinct, but there are differences among various processed RC. The different samples of processed RC were classified into two and the result of hierarchical clustering is correlated with the property of traditional Chinese drug. CONCLUSION: It is proved that the method which is convenient, rapid and repeat well can be used to identify different samples of processed RC. The change of property of different processed RC is related with processing adjuvant, the possible mechanism is that new components are produced or the contents of known components have changed due to the adjuvant during processing.

[Probing into objectivity of adverse drug reaction of traditional Chinese medicine].

The adverse drug reaction (ADR) of traditional Chinese medicine (TCM) has been concerned nowadays. Some bans were published to limit their use abroad. There wasn't a feasible criterion to reply to the ADR of TCM yet in China. The ADR of 6 kinds of typical TCM which state attached importance to was chosen to discuss. Based on the retrospect investigation, the objectivity of ADR was explored in this study.

[Effects of different extracts from Radix isatidis on lymphocytes of mice by biothermodynamics].

OBJECTIVE: To observe the different extracts from Radix Isatidis on multiplication of mice lymphocytes. METHOD: Lymphocytes were separated and cultured. Immunological activities of different extracts from Radix Isatidis were studied by thermodynamics and the results were tested by the conventional pharmacological experiments. RESULT: The results showed that the water extract and residue had significant immunological effects while organic solvent extracts had immunological activity to some extent. CONCLUSION: The comparison of immunological activity among the extracts from Radix Isatidis were as follows: residue after extracting > general extract > nBuOH extract > EtOAc extract > CHCl3 extract > P E extract.