Liwei Guo

Comparative pharmacokinetics of baicalin in plasma after oral administration of Huang-Lian-Jie-Du-Tang or pure baicalin in MCAO and sham-operated rats.

A sensitive and specific HPLC method was developed to analyze baicalin in rat plasma. The author had compared the pharmacokinetics of baicalin after oral administration of HLJDT decoction or pure baicalin in MCAO and sham-operated rats. All the rats were divided into two groups, MCAO and sham-operated rats. Each group contained two subgroups: HLJDT decoction and pure baicalin subgroup. The HLJDT subgroup oral administration of HLJDT decoction extract 10.00 g/kg according to body weight (containing baicalin 400.00 mg/kg according to body weight), the pure baicalin subgroup received a gavages at a dosage of baicalin 400.00 mg/kg according to body weight too. The pharmacokinetics parameters were analyzed by kinetica. The results indicated that the pharmacokinetics of baicalin in rat plasma was non-linear and there were significant differences between different groups. No matter in MCAO or sham-operated rats, pure baicalin had shown better absorption than HLJDT decoction. Whether administration of pure baicalin or HLJDT decoction, the MCAO rats show better, quicker absorption of baicalin than sham-operated rats. It was good for baicalin to exert pharmacological effects on healed cerebrovascular diseases. The method had been applied successfully to pharmacokinetics of baicalin in rat plasma after oral administration of pure baicalin or HLJDT decoction.

Systematic review of recent advances in pharmacokinetics of four classical Chinese medicines used for the treatment of cerebrovascular disease

Recent studies have focused more on Chinese medicine used for the treatment of cerebrovascular disease. The current review covers researches on the pharmacokinetics of Chinese medicine, providing a convenient reference for researchers to increase efficiency of drug discovery, by compiling and discussing the pharmacokinetics of four classical Chinese medicines for therapy of cerebrovascular disease containing: Panax notoginseng, Salvia miltiorrhiza, Ligusticum Chuanxiong and Gardenia. It also helps to eliminate side effect as far as possible from inappropriate Chinese medicine usage. Current integrative and comprehensive review of Chinese medicine for cerebrovascular disease including 1) the absorption of some constituents is limited such as ginsenosides Rg1 and Rb1. It may be affected by gastric juice, first-pass effect, etc. 2) The interactions between Chinese medicine and prescription can occur. Borneol and carbomer would enhance the absorption of R1 and Rg1 in vivo by increasing adjacent cell transport ability. 3) The distribution of active constituents in brain is important for cerebrovascular disease. BBB protects brain from xenobiotic. Intranasal, intra-tympanic administration is a promising alternative to conventional administration to reach brain for ligustrazine. 4) Renal excretion is the uppermost route of these Chinese medicines. But biliary, fecal and urinary excretion are the other major routes. Theoretical and practical aspects are described with pharmacokinetic examples. In the end, this paper also discusses recent development of bio-analysis of Chinese medicine.

Integrated pharmacokinetics of major bioactive components in MCAO rats after oral administration of Huang-Lian-Jie-Du-Tang.

ETHNOPHARMACOLOGICAL RELEVANCE Huang-Lian-Jie-Du-Tang (HLJDT, or Oren-gedoku-to in Japanese), an important multi-herb remedy in China and other Asia countries, has been used clinically to treat cerebral ischemia for decades. MATERIALS AND METHODS According to the previous studies we have reported, an HPLC method was developed and validated for determination of berberine, palmatine, baicalin, baicalein and geniposide simultaneously in MCAO rat plasma after administration of HLJDT aqueous extract. A classified integral pharmacokinetic method was put forward after having compared the integrated concentration-time profile with that of single component. An AUC based weighting approach was used for integrated principle. RESULTS The results indicated the classified integral pharmacokinetic profile of index components from HLJDT could reveal the pharmacokinetic behavior of original components, and was corresponding to the holistic pharmacological effects of anti-ischemia with HLJDT. CONCLUSIONS This study was aimed to explore an approach that could be applied to integrate the pharmacokinetic behavior of different components derived from HLJDT. The integrated pharmacokinetic results also provided more information for further understanding of the clinical cerebrovascular disease in use of HLJDT.

Immuno-enhancement effects of ethanol extract from Cyrtomium macrophyllum (Makino) Tagawa on cyclophosphamide-induced immunosuppression in BALB/c mice.

ETHNOPHARMACOLOGICAL RELEVANCE Cyrtomium macrophyllum (Makino) Tagawa has been traditionally used as a herbal medicine for the treatment of various infectious diseases such as tapeworm infestation, colds, and viral diseases. However, no systematic study of the immunity of Cyrtomium macrophyllum ethanol extracts (CM) has yet been reported. The present work evaluates these traits. MATERIALS AND METHODS 120 male BALB/c mice were divided into 6 groups of 20 mice each: (1) normal group (sterile physiological saline), which served as a blank control; (2) model group (Cyclophosphamide, CY) group (sterile physiological saline), which served as a negative control; (3) low-dose CM (50 mg/kg BW); (4) intermediate-dose CM (100 mg/kg BW); (5) high-dose CM (200 mg/kg BW); (6) CM group (200 mg/kg BW). CY (0.2 ml) was administered via intraperitoneal injection. The other regimens were administered via gavage in 0.2 ml solution. Phytochemical of CM was characterized by HPLC-LTQ-Orbitrap. The acute toxicity effect of the ethanol extract of Cyrtomium macrophyllum was also investigated. RESULTS The spleen and thymus indices of mice receiving low, intermediate, and high doses of CM recovered more quickly than those of CY mice, and they did so in a dose-dependent manner. These mice also showed higher T cell and B cell proliferation responses and macrophage function than those of CY mice, and their serum levels of interleukin-6 and interferon-γ had become normal. In acute toxicity test, CM exhibited no mortality and behavioral changes in mice. Quantitative phytochemical analysis showed flavonoids, polyphenols, and tannins to be the major compounds present in the extract, at 27.64%, 30.87%, and 11.22%, respectively. We found that 16 compounds were characterized by the interpretation of their mass spectra obtained by the MS/MS. CONCLUSION The current study demonstrates that Cyrtomium macrophyllum ethanol extract improved immune function in CY-treated mice.

Preconditioning with the traditional Chinese medicine Huang-Lian-Jie-Du-Tang initiates HIF-1α-dependent neuroprotection against cerebral ischemia in rats

ETHNOPHARMACOLOGICAL RELEVANCE Huang-Lian-Jie-Du-Tang (HLJDT) is a classical heat-clearing and detoxicating formula of traditional Chinese medicine that is widely used to treat stroke. The present study was designed to investigate the effects of HLJDT preconditioning on neurons under oxygen and glucose deprivation (OGD) and rats subjected to middle cerebral artery occlusion (MCAO). MATERIALS AND METHODS A stroke model of rats was obtained through MCAO. Following HLJDT preconditioning, the cerebral infarction volume, cerebral water content, and neurological deficient score were determined. Cerebral cortical neurons cultured in vitro were preconditioned with HLJDT and then subjected to OGD treatment. The release of lactate dehydrogenase (LDH) from neurons was detected. The levels of hypoxia-inducible factor-1α (HIF-1α) and PI3K/Akt signaling were analyzed by western blotting, and the levels of erythropoietin (EPO) and vascular endothelial growth factor (VEGF) in the supernatant of the neurons and the plasma of MCAO rats were measured through a radioimmunological assay. The apoptosis and proliferation of neurons were analyzed by immunohistochemistry. RESULTS HLJDT preconditioning significantly reduced the cerebral infarction volume and cerebral water content and ameliorated the neurological deficient score of MCAO rats. In addition, HLJDT preconditioning protected neurons against OGD. Increased HIF-1α, EPO, and VEGF levels and the activation of PI3K/Akt signaling were observed as a result of HLJDT preconditioning. Furthermore, HLJDT preconditioning was found to inhibit ischemia-induced neuron apoptosis and to promote neuron proliferation under conditions of ischemia/reperfusion. CONCLUSION Both rats and neurons subjected to HLJDT preconditioning were able to resist ischemia/reperfusion or hypoxia injury through the inhibition of apoptosis and the enhancement of proliferation, and these effects were primarily dependent on the activation of the PI3K/Akt signaling pathway and HIF-1α.

Distribution of α-asarone in brain following three different routes of administration in rats.

The goal of the present paper is to compare the distributions of α-asarone administered to rats through three different routes: oral, intravenous and intranasal. The concentrations of α-asarone in seven distinct brain regions, the olfactory bulb, cerebellum, hypothalamus, frontal cortex, striatum, hippocampus and medulla/pons as well as in plasma and cerebrospinal fluid (CSF), were determined by HPLC. The quantities of α-asarone accumulated in liver were measured to determine whether α-asarone could generate hepatotoxicity when administered via the three different routes. The results indicated that α-asarone could be absorbed via two different routes into the brain, after intranasal administration of dry powders. In the systemic route, α-asarone immediately entered the brain through the blood-brain barrier (BBB) after uptake into the circulatory system. In the olfactory bulb route, α-asarone traveled from the olfactory epithelium in the nasal cavity straight into brain tissue via the olfactory bulb. Furthermore, intranasal administration of α-asarone as a dry powder can ensure quick absorption and avoid excessive concentrations in the blood and liver, while achieving concentrations in the brain comparable to those attained by intravenous and oral administration routes.

Increased involvement of Panax notoginseng in the mechanism of decreased hepatotoxicity induced by Tripterygium wilfordii in rats.

ETHNOPHARMACOLOGICAL RELEVANCE The key problem with toxic Chinese herbs in clinical applications is how to find the most effective method to reduce toxicity. This study focuses on discussing the mechanism of decreased hepatotoxicity by the usage compatibility of two commonly used traditional Chinese drugs that are used clinically: Tripterygium wilfordii Hook. f. (TW) and Panax notoginseng (Burkill) F.H. Chen (PN). Additionally, based on the results from using metabonomics technology, the usage compatibility with these two herbs that was originated from clinical experience is the first to clarify the rationality of the drug combination. MATERIALS AND METHODS Through a fast and effective HPLC method, plasma concentration-time profiles and triptolide distribution characteristics in liver, heart, spleen, lung and kidney tissues were simultaneously determined in rats after oral administration of the aqueous extract of TW and TW-PN. The reduced hepatotoxicity data of the usage compatibility with TW and PN were also investigated, and then a UHPLC-QTOF/MS method was developed and validated for the explanation of the reduced hepatotoxicity mechanism. RESULTS It was indicated that nine endogenous metabolites might be potential biomarkers for hepatotoxicity induced by TW. In addition, the plasma concentration-time profiles and the distribution characteristics of TP in rats were changed after oral administration of the aqueous extract of TW-PN, and simultaneously, the hepatotoxicity was obviously decreased. CONCLUSIONS The results indicated that usage compatibility with TW and PN was reasonable in clinical use. To the best of our knowledge, this is the first report to describe the mechanism of reducing hepatotoxicity with the combined use of TW and PN from clinical experience.

Berberine Preconditioning Protects Neurons Against Ischemia via Sphingosine-1-Phosphate and Hypoxia-Inducible Factor-1α

Berberine exerts neuroprotective and modulates hypoxia inducible factor-1-alpha (HIF-1[Formula: see text]. Based on the role of HIF-1[Formula: see text] in hypoxia preconditioning and association between HIF-1[Formula: see text] and sphingosine-1-phosphate (S1P), we hypothesized that berberine preconditioning (BP) would ameliorate the cerebral injury induced by ischemia through activating the system of HIF-1[Formula: see text] and S1P. Adult male rats with middle cerebral artery occlusion (MCAO) and rat primary cortical neurons treated with oxygen and glucose deprivation (OGD) with BP at 24[Formula: see text]h (40[Formula: see text]mg/kg) and 2[Formula: see text]h (10[Formula: see text][Formula: see text]mol/L), respectively, were used to determine the neuroprotective effects. The HIF-1[Formula: see text] accumulation, and S1P metabolism were assayed in the berberine-preconditioned neurons, and the HIF-1[Formula: see text]-mediated transcriptional modulation of sphingosine kinases (Sphk) 1 and 2 was analyzed using chromatin immunoprecipitation and real-time polymerase chain reaction. BP significantly prevented cerebral ischemic injury in the MCAO rats at 24[Formula: see text]h and 72[Formula: see text]h following ischemia/reperfusion. In OGD-treated neurons, BP enhanced HIF-1[Formula: see text] accumulation with activation of PI3K/Akt, and induced S1P production by activating Sphk2 via the promotion of HIF-1[Formula: see text]-mediated Sphk2 transcription. In conclusion, BP activated endogenous neuroprotective mechanisms associated with the S1P/HIF-1 pathway and helped protect neuronal cells against hypoxia/ischemia.

Study on the Absorption Mechanism of Geniposide in the Chinese Formula Huang-Lian-Jie-Du-Tang in Rats

Huang-Lian-Jie-Du-Tang (HLJDT) is a classical recipe for relieving fever and toxicity for thousands of years in China. Geniposide is one of the main components in HLJDT. The present study was conducted in order to investigate the differences of absorption of geniposide after oral administration of geniposide alone and HLJDT in rats. Pharmacokinetic differences of geniposide following oral administrations of pure geniposide and HLJDT were investigated in vivo. The absorption of geniposide in pure compound and HLJDT was evaluated using intestinal perfusion and Caco-2 models. The in vivo and in vitro studies showed good relevance and consistent results. The co-occurring components in HLJDT were found to promote the absorption of geniposide from the pharmacokinetic study in vivo, intestinal perfusion, and Caco-2 model. Geniposide had better absorption in the duodenum and jejunum from the intestinal perfusion model, which was mainly absorbed by passive diffusion. Verapamil influenced the transportation of geniposide, while EDTA did not, demonstrating that geniposide might be the potential substance of P-glycoprotein in intestinal perfusion and Caco-2 models. The absorption of geniposide was studied systematically to guide the design of the oral dosage of geniposide and HLJDT in clinical therapy.

Purification and biochemical characterization of a novel fibrinolytic enzyme, PSLTro01, from a medicinal animal Porcellio scaber Latreille.

A novel protease, named PSLTro01, with fibrinolytic and anticoagulant activity was isolated from Porcellio scaber Latreille and was purified by a combination of hollow fibre membrane molecular weight cut-off (MWCO), ammonium sulfate fractionation, gel filtration and ion-exchange chromatography. PSLTro01 is a single-chain protein with a molecular mass of 38,497 Da as estimated by non-reduced SDS-PAGE and MALDI-TOF MS spectrometry, and its N-terminal 15 amino acid sequence was determined as DINGGGATLPQPLYQ. PSLTro01 is stable in the range of 20-40 °C and pH 6.0-10.0, with a maximum fibrinolytic activity at 40 °C and pH 7.0. The PSLTro01-induced fibrinolytic activity was not influenced by K(+) or Na(+) but was slightly increased by Mg(2+) and completely inhibited by aprotinin and pepstatin A. Fibrin plate assays revealed that PSLTro01 could not directly degrade fibrin but was a plasminogen activator. PSLTro01 exhibited high specificity for the substrate S-2251 for plasmin, followed by S-2238 for thrombin and S-2444 for urokinase. Moreover, the fibrinogenolysis pattern of PSLTro01 was Aα-chains>Bβ-chains>γ-chain. Tail-thrombus of the enzyme treated group was significantly shorter than the physiological saline treated group and the thrombus decrement was correlated with the enzyme dose. PSLTro01 prolongs both thrombin time (TT) and activated partial thromboplastin time (APTT). These results indicate that PSLTro01 may have potential applications in the prevention and treatment of thrombosis.