Traditional Chinese medicine's intervention in endothelial nitric oxide synthase activation and nitric oxide synthesis in cardiovascular system.

Cardiovascular disease (CVD) is one of the most dangerous diseases which has become a major cause of human death. Many researches evidenced that nitric oxide (NO)/endothelial nitric oxide synthase (eNOS) system plays a significant role in the occurrence and development of CVD. NO, an important signaling molecule, closely associated with the regulation of vasodilatation, blood rheology, blood clotting and other physiological and pathological processes. The synthesis of NO in the endothelial cells primarily depends on the eNOS activity, thus the exploration of the mechanisms and effects of the eNOS activation on NO production is of great significance. Recently, studies on the effects of traditional Chinese medicine (TCM) and its extracts on eNOS activation and NO synthesis have gradually attracted more and more attentions. In this paper, we reviewed the mechanisms of NO synthesis and eNOS activation in the vascular endothelial cells (VECs) and intervention of TCM, so as to provide reference and train of thought to the intensive study of NO/eNOS system and the research and development of new drug for the treatment of CVD.

Effective dose of the combination of salvianolic acids and tanshinones against inflammatory injury in human umbilical vein endothelial cells injured by thrombin.

OBJECTIVE: To investigate the pharmacological effective doses of Chinese Danshen components, (salvianolic acids and tanshinones) combinations on vascular endothelia cells against inflammatory injury. METHODS: Thrombin (50 U/mL) was incubated in human umbilical vein endothelia cells (HUVECs) with 50 and 100 µg/L (1:0, 0:1, 3:1, 2:1, 1:1, and 1:2) salvianolic acids and tanshinones or without the components of Danshen extract. Cell viability was confirmed in (4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium (MTT) assay and cell death was confirmed in lactate dehydrogenase (LDH) released into the cell culture supernatant. RESULTS: Six hours treatment of HUVECs with 50 or 100 µg/L salvianolic acids and tanshinones combinations at ratios 1:0, 0:1, 3:1, 2:1, 1:1, and 1:2 significantly (P<0.05 or P<0.01) improved cell proliferation in vitro compared with model group when assayed in MTT, whereas 100 µg/L groups displayed trend toward upshift in potency compared with 50 µg/L groups (2:1, 1:1, and 1:2), respectively, in addition to inhibition of cell death confirmed in reduced LDH released into the culture medium. CONCLUSIONS: These findings provided some useful information for safe and effective use of Danshen preparations in clinical practice. We made observations regarding the protective effect of Danshen components combinations in vascular endothelial against inflammatory injury caused by thrombin activation of endothelial cell.

[Network analysis of ethanol precipitation process for Schisandrae chinensis fructus].

A set of central composite design experiments were designed by using four factors which were ethanol amount, ethanol concentration, refrigeration temperature and refrigeration time. The relation between these factors with the target variables of the retention rate of schizandrol A, the soluble solids content, the removal rate of fructose and the removal rate of glucose were analyzed with Bayesian networks, and ethanol amount and ethanol concentration were found as the critical process parameters. Then a network model was built with 2 inputs and 4 outputs using back propagation artificial neural networks which was optimized by genetic algorithms. The R2 and MSE from the training set were 0.983 8 and 0.001 1. The R2 and MSE from the test set were 0.975 9 and 0.001 8. The results showed that network analysis method could be used for modeling of Schisandrae Chinensis Fructus ethanol precipitation process and identify critical operating parameters.

[Reliability theory based on quality risk network analysis for Chinese medicine injection].

A new risk analysis method based upon reliability theory was introduced in this paper for the quality risk management of Chinese medicine injection manufacturing plants. The risk events including both cause and effect ones were derived in the framework as nodes with a Bayesian network analysis approach. It thus transforms the risk analysis results from failure mode and effect analysis (FMEA) into a Bayesian network platform. With its structure and parameters determined, the network can be used to evaluate the system reliability quantitatively with probabilistic analytical appraoches. Using network analysis tools such as GeNie and AgenaRisk, we are able to find the nodes that are most critical to influence the system reliability. The importance of each node to the system can be quantitatively evaluated by calculating the effect of the node on the overall risk, and minimization plan can be determined accordingly to reduce their influences and improve the system reliability. Using the Shengmai injection manufacturing plant of SZYY Ltd as a user case, we analyzed the quality risk with both static FMEA analysis and dynamic Bayesian Network analysis. The potential risk factors for the quality of Shengmai injection manufacturing were identified with the network analysis platform. Quality assurance actions were further defined to reduce the risk and improve the product quality.

[Data integration, data mining and visualization analysis of traditional Chinese medicine manufacturing process].

Huge amount of data becomes available from the pharmaceutical manufacturing process with wide application of in- dustrial automatic control technology in traditional Chinese medicine (TCM) industry. The industrial big data thus provides golden op- portunities to better understand the manufacturing process and improve the process performance. Therefore it is important to implement data integration and management systems in TCM plants to easily collect, integrate, store, analyze, communicate and visulize the data with high efficiency. It could break the data island and discover useful information and knowledge to improve the manufacturing process performance. The key supporting technologies for TCM manufacturing and industrial big data management were introduced in this paper, with a specific focus on data mining and visualization technologies. Using historic data collected from a manufacturing plant of Shengmai injection of SZYY group, we illustrated the usefulness and discussed future prospects of data mining and visualization technologies.

[Multi-objective optimization of extraction process for red ginseng based upon extraction efficiency and cost control].

It is the objective of this study to optimize the extraction process of red ginseng to minimize the unit cost of extracting effective ingredients. The relation between the target variables of total quantity of ginsenosides and first extraction time, first extraction solution amount, second extraction time, second extract solution amount were studied with Box-Behnken experimental design method. At the same we also considered the cost of extraction solution and energy usage. The objective function was set as unit cost of target (total quantity of ginsenosides or its purity) for the multi-objective optimization of extraction process. As a result, the optimal process parameters were found as first extraction time (108.7 min), first extraction solution amount folds (12), second extraction time (30 min), second extraction solution amount folds (8) to minimize the unit cost. It indicated that this approach could potentially be used to optimize industrial extraction process for manufacturing Chinese medicine.

[Dynamic predictive modeling of extraction process for red ginseng using near-infrared spectroscopy].

It is the objective of this study to develop dynamic predictive model for the extraction process of red Ginseng using NIR spectroscopy. NIR spectroscopy was collected online and PLSR models were developed for total quantity of ginsenosides. The performance of NIR prediction model achieved R, RMSEC, RMSEP of 0.996 09, 0.018 9, 0.016 8, respectively. A first order dynamic mass transfer model was combined with NIR prediction of the quality indicator to predict the trajectory of the extraction process based upon the initial 3 or 4 data points. The results showed good agreement with actual measurements indicating reasonable accuracy of the predictive model. It could potentially be used for advanced predictive control of the extraction process.

[Strategies and key technologies for risk control and management in quality of Chinese medicine injections based on integrated pharmacology].

This paper focuses on the quality risk control and management of Chinese medicine (CM) injections. The most important technological requirements are analyzed, and a strategy for integrated pharmacology to study CM mechanism is proposed. A key technology system for quality risk control and management was further constructed. The strategy and technology system was finally applied to Shengmai injection for quality risk control and management.

[Progress on chemical components and anti-cerebral injury effects of storax].

The Chinese medicine storax is one of the resuscitation-inducing aromatic herbs used to treat stoke, epilepsy, convulsion, etc. In this review, authors summarized the progress on chemical compositions and anti-cerebral injury effects of storax. Gas chromatography-mass spectrometry (GC-MS) were used to determine the main chemical compositions of storax . And the anti-cerebral ischemia, anti-convulsion, and anti-memory defect effects of storax and its compound preparation were discussed. At last, authors have tried to propose a number of recommendations for the future research.

Cerebrospinal fluid pharmacology: an improved pharmacology approach for chinese herbal medicine research.

Despite many successful applications of Chinese herbal medicine (CHM) in the treatment and prevention of neurological diseases (ND), the fully scientific understanding of CHM's action mechanisms had been hampered for lack of appropriate methods to explore the combinatorial rules, the synergistic mechanisms, and the molecular basis of CHM. As an improved pharmacology approach, cerebrospinal fluid pharmacology (CSFP), based on the fact that cerebrospinal fluid plays an important role in the health maintenance of specific survival environment for neurons and glial cells, has been constructed and applied to CHM research for treating ND. In the present review, the concept and advantages of CSFP are briefly introduced. The approaches and key technologies of CSFP in CHM research are also collated and analyzed. Furthermore, the developing tendency of CSFP is summarized, and its framework in CHM research is also proposed. In summary, CSFP provides a new strategy not only to eliminate some barriers of CHM research for treating ND, but also to broaden the pharmacology research for bridging the gap between CHM and modern medicine. Moreover, the advancements in CSFP will bring about a conceptual move in active ingredients discovery of CHM and make a significant contribution to CHM modernization and globalization.

Characterization of in vivo antioxidant constituents and dual-standard quality assessment of Danhong injection.

Antioxidants and oxidative stress play a critical role in cardiovascular diseases. Danhong injection (DHI) is a well prescribed cardiovascular medication in China, but its detailed chemical basis and mechanisms of action remain unknown. To prove the antioxidant activity of DHI, its free radical scavenging capacity (RSC) was assessed by 2,2-diphenyl-1-picrylhydrazyl (DPPH) spectrophotometric assay. The 50% radical scavenging activity value was 1:129.2 mL/mL, against 0.95 mM DPPH. To further identify the antioxidant compounds, modified thin-layer chromatography combined with DPPH bioautography assay was used. Compared with vitamin C, 11 of 16 available compounds displayed strong antioxidant activity, which were also detected in rat serum after intravenous administration of DHI by ultra-performance liquid chromatography-tandem mass spectrometry, except for hydroxysafflor yellow A. Therefore, 10 antioxidants remaining in the blood as key markers, and six other compounds as general markers, were employed to perform the quality control of DHI by ultra-performance liquid chromatography-ultraviolet detection after systematic methodological validation. The analytical results indicate a high correlation (r = 0.9) between the total content of those antioxidants remaining in blood and RSC of DHI among 10 batches. Further, the antioxidant profiling and chemical marker quantification as dual-standard quality assessment was successfully applied to evaluate Danshen and safflower injections.

Caffeic acid ester fraction from Erigeron breviscapus inhibits microglial activation and provides neuroprotection.

OBJECTIVE: To investigate the effects of caffeic acid ester fraction (Caf) from Erigeron breviscapus, mainly composed of dicaffeoylquinic acids (diCQAs), on microglial activation in vitro and focal cerebral ischemia in vivo. METHODS: The production of nitric oxide (NO), tumor necrosis factor α (TNF-α), and interleukin-1ß (IL-1ß) induced by lipopolysaccharide (LPS) treatment in rat primary cultured microglia were measured by Griess reaction or enzyme-linked immunosorbent assay. Cell viability of cortical neurons was measured using AlamarBlue reagent. The behavioral tests and the infarct area of brain were used to evaluate the damage to central nervous system in rat middle cerebral artery occlusion (MCAO) model of cerebral ischemia. Real time polymerase chain reaction was used to determine the expression of inducible nitric oxide synthase (iNOS), TNF-α and IL-1ß mRNA in ischemic cerebral tissues. RESULTS: Caf inhibited the production of NO, TNF-α and IL-1ß induced by LPS treatment in primary microglia in a dose-dependent manner. Exposure of cortical neurons to conditioned medium from Caf-treated microglia increased neuronal cell viability (P<0.01) compared with conditioned medium from LPS-treated alone. In MCAO rat model of cerebral ischemia, Caf could significantly improve neurobehavioural performance and reduce percentage infarct volume compared with the vehicle group (P<0.05). Caf could also significantly inhibit the up-regulation of iNOS, TNF-α, and IL-1ß gene expressions in ischemic cerebral tissues. CONCLUSION: Caf could suppress microglial activation, which may be one mechanism of its neuroprotective effect against ischemia.

Chinese Medicine's Intervention Effect on Nogo-A/NgR.

Cerebral vascular disease is very common in the elderly and is one of the most dangerous diseases which is hazardous to the body's health, and it is the medical specialists' study hot spot not only in the clinical field but also in the medical basic research field. Neural regeneration has been paid more and more attention in recent years. Nogo's function in the process of neural regeneration has become the focal point since it was discovered in the year 2000. Many studies elucidate that Nogo negatively affects the neural regeneration and plasticity. Chinese medicine plays an important role in the prevention and treatment of neural diseases, and recently some researches about the Chinese medicine's intervention effect on Nogo-A/NgR sprang up, so it is necessary to make a review on this aspect.

Neuroprotective effects of scutellarin against hypoxic-ischemic-induced cerebral injury via augmentation of antioxidant defense capacity.

An increasing number of studies has indicated that hypoxic-ischemic-induced cerebral injury is partly mediated via oxidative stress. Recent researches have focused on searching for drug and herbal manipulations to protect against hypoxic-ischemic-induced oxidative cell damage. Scutellarin is a flavonoid derived from the Erigeron breviscapus (vant.) and has been reported to exhibit neuroprotective properties. However, its precise mechanism, particularly its antioxidation mechanism, remains elusive. In the present study, we investigated the neuroprotective effects of scutellarin on middle cerebral artery occlusion (MCAO)-induced brain damage in rats, and oxygen-glucose deprivation (OGD)-induced toxicity in primary culture of rat cortical neurons. In vivo, intraperitoneal injections of scutellarin (20 and 60 mg/kg) improved the neurological score and diminished the percentage of brain infarct volume. At the same time, scutellarin significantly increased superoxide dismutase (SOD), catalase (CAT) activities and glutathione (GSH) level in ischemic brain tissues, enhancing endogenous antioxidant activity. Moreover, pretreatment of scutellarin (25, 50 and 100 µM) protected neurons against lethal stimuli, decreased the percentage of apoptotic cells and inhibited reactive oxygen species (ROS) generation in OGD-induced primary cortical neurons in vitro. These results suggest that the preventive and therapeutic potential of scutellarin in cerebral injury patients is, at least in part, ascribed to augmentation of cellular antioxidant defense capacity.

In vivo pharmacokinetics of bakuchiol after oral administration of bakuchiol extraction in rat plasma.

ETHNOPHARMACOLOGICAL RELEVANCE: Psoralea corylifolia L. (Fabaceae) was a traditional Chinese medicine for the treatment of various skin diseases such as psoriasis, vitiligo and chronic graft-versus-host, and has been proved to show anticancer, cytotoxic, anti-bacterial, cardiac, diaphoretic, diuretic, stimulant, aphrodisiac and tonic effects. Bakuchiol was one of the main active ingredients of this traditional Chinese medicine. AIM OF THE STUDY: In this paper, pharmacokinetic study was conducted to obtain pharmacokinetic parameters of bakuchiol. MATERIALS AND METHODS: Bakuchiol was enriched using resin inform the ethanol extract of Psoralea corylifolia L., HPLC-UV was used to determine the concentration of bakuchiol in rat plasma at different time points after administration. The main pharmacokinetic parameters of bakuchiol in rat were obtained based on the analysis of the plasma sample. RESULTS: The pharmacokinetics of bakuchiol was fitted with a two-compartment model and it was eliminated relative slowly in rats. CONCLUSIONS: The HPLC-UV method was successfully applied to study the pharmacokinetics of bakuchiol in rats.

Potency fingerprint of herbal products Danshen injection for their quality evaluation.

The fingerprint technique has been studied frequently as a useful strategy for quality of traditional Chinese medicine. A novel potency fingerprint that can quantitatively analyze the antioxidant activity of individual constituent and provide the total antioxidant activities of the samples has been developed by high-performance liquid chromatography coupled with ultraviolet and pyrogallol-luminol chemiluminescence detection (HPLC-diode array detection (DAD)-PLD). Hierarchical clustering analysis has been used as a powerful pattern recognition tool to identify and classify Danshen injection from different factories. In addition, the combination use of the chromatographic fingerprint and potency fingerprint with principal component analysis was applied to quality control of Danshen injection. The results demonstrated that the proposed potency fingerprint was a useful means to control the quality and to clarify the possible mechanism of action of herbal products.

The anti-inflammatory activities of Tanshinone IIA, an active component of TCM, are mediated by estrogen receptor activation and inhibition of iNOS.

Tanshinone IIA (Tan IIA) is a major compound extracted from a traditional herbal medicine Salvia miltiorrhiza BUNGE, which is used to treat cardiovascular diseases, cerebrovascular diseases and postmenopausal syndrome. It has also been shown to possess anti-inflammatory activity. Since Tan IIA has a similar structure to that of 17beta-estradiol (E(2)), the present study was undertaken to characterize the estrogenic activity of Tan IIA and to demonstrate a functional role of this activity in RAW 264.7 cells. In transient transfection assay, Tan IIA (10 microM) increases ERE-luciferase activity in an estrogen receptor (ER) subtype-dependent manner when either ERalpha or ERbeta were co-expressed in Hela cells. In LPS-induced RAW 264.7 cells, Tan IIA exerts anti-inflammatory effects by inhibition of iNOS gene expression and NO production, as well as inhibition of inflammatory cytokine (IL-1beta, IL-6, and TNF-alpha) expression via ER-dependent pathway. Therefore, it could serve as a potential selective estrogen receptor modulator (SERM) to treat inflammation-associated neurodegenerative and cardiovascular diseases without increasing the risk of breast cancer.

The antioxidant-activity-integrated fingerprint: an advantageous tool for the evaluation of quality of herbal medicines.

The activity-integrated fingerprints of Danshen injections have been established by high-performance liquid chromatography coupled with chemiluminescence detection (HPLC-CL) according to their antioxidants activity scavenging hydrogen peroxide. The authentication and validation of the activity-integrated fingerprints were performed. Then, a data-level information fusion method was employed to capture the chemical and antioxidant information encoded in HPLC-CL fingerprints. Finally, based on the fusion results, the quality of different batches of Danshen injections samples were further evaluated by similarity measure. In comparison with conventional fingerprints, the activity-integrated fingerprints, which simultaneously contained the chemical characteristics and antioxidant activities of constitutions could comprehensively and properly reveal the quality characteristics of the Danshen injections. In conclusion, the antioxidant-activity-integrated fingerprints were suitable for quality control of Danshen injections. This study could clearly demonstrate that the activity-integrated fingerprint was a powerful and meaningful tool to comprehensively improve the quality control of complex herbal medicines.

[Effect of dan-shen, san-qi of different proportion on platelet aggregation and adhesion in normal rabbits].

OBJECTIVE: To observe the effect of the crude extracts of Dan-shen root and San-qi of different proportion on platelet aggregation and adhesion in normal rabbits. METHOD: With rabbits, ig. (4d, exsanguinated via carotid artery, percentage of platelet aggregation and adhesion was measured. RESULT AND CONCLUSION: Aspirin (4.4 mg/Kg) could markedly inhibit platelet aggregation and adhesion in normal rabbit. The proportions of Dan-shen root/San-qi (10:0, 10:1, 10:3, 10:6, 1:10) could markedly inhibit platelet aggregation, among which 10:3 was the best. San-qi alone had little effect on aggregation. The proportions of Dan-shen root/San-qi (10:3, 10:6, 0:10) could markedly inhibit platelet adhesion, among which 0:10 was the best, and the proportions(10:0, 10:1, 1:10) had little effect.