Synergistic amelioration between Ligusticum striatum DC and borneol against cerebral ischemia by promoting astrocytes-mediated neurogenesis.

ETHNOPHARMACOLOGICAL RELEVANCE: Ligusticum chuanxiong Hort (LCH), with the accepted name of Ligusticum striatum DC in "The Plant List" database, is a widely used ethnomedicine in treating ischemic stroke, and borneol (BO) is usually prescribed with LCH for better therapy. Our previous study confirmed their synergistic effect on neurogenesis against cerebral ischemia. However, the underlying mechanism is still unclear. AIM OF THE STUDY: More and more evidence indicated that astrocytes (ACs) might be involved in the modulation of neurogenesis via polarization reaction. The study was designed to explore the synergic mechanism between LCH and BO in promoting astrocyte-mediated neurogenesis. MATERIALS AND METHODS: After primary cultures and identifications of ACs and neural stem cells (NSCs), the oxygen-glucose deprivation (OGD) model and the concentrations of LCH and BO were optimized. After the OGD-injured ACs were treated by LCH, BO, and their combination, the conditioned mediums were used to culture the OGD-injured NSCs. The proliferation, migration, and differentiation of NSCs were assessed, and the secretions of BDNF, CNTF, and VEGF from ACs were measured. Then the expressions of C3 and PTX3 were detected. Moreover, the mice were performed a global cerebral ischemia/reperfusion model and treated with LCH and (or) BO. After the assessments of Nissl staining, the expressions of Nestin, DCX, GFAP, C3, PTX3, p65 and p-p65 were probed. RESULTS: The most appropriate duration of OGD for the injury of both NSCs and ACs was 6 h, and the optimized concentrations of LCH and BO were 1.30 µg/mL and 0.03 µg/mL, respectively. The moderate OGD environment induced NSCs proliferation, migration, astrogenesis, and neurogenesis, increased the secretions of CNTF and VEGF from ACs, and upregulated the expressions of C3 and PTX3. For the ACs, LCH further increased the secretions of BDNF and CNTF, enhanced PTX3 expression, and reduced C3 expression. Additionally, the conditioned medium from LCH-treated ACs further enhanced NSC proliferation, migration, and neurogenesis. The in vivo study showed that LCH markedly enhanced the Nissl score and neurogenesis, and decreased astrogenesis which was accompanied by downregulations of C3, p-p65, and p-p65/p65 and upregulation of PTX3. BO not only decreased the expression of C3 in ACs both in vitro and in vivo but also downregulated p-p65 and p-p65/p65 in vivo. Additionally, BO promoted the therapeutic effect of LCH for most indices. CONCLUSION: A certain degree of OGD might induce ACs to stimulate the proliferation, astrogenesis, and neurogenesis of NSCs. LCH and BO exhibited a marked synergy in promoting ACs-mediated neurogenesis and reducing astrogenesis, in which LCH played a dominant role and BO boosted the effect of LCH. The mechanism of LCH might be involved in switching the polarization of ACs from A1 to A2, while BO preferred to inhibit the formation of A1 phenotype via downregulating NF-κB pathway.

Exploring the synergic mechanism of Ligusticum striatum DC. and borneol in attenuating BMECs injury and maintaining tight junctions against cerebral ischaemia based on the HIF-1α/VEGF signalling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Ligusticum striatum DC., also known as Ligusticum chuanxiong Hort. (LCH), is widely used in China for its excellent effect in ischaemic stroke (IS) patients, and borneol (BO) has been confirmed to maintain the blood‒brain barrier (BBB) after stroke. They are often used as a combination in the prescriptions of IS patients. Although the advantage of their combined treatment in improving brain ischaemia has been verified, their synergistic mechanism on BBB maintenance is still unclear. AIM OF THE STUDY: This study was designed to evaluate the synergistic effect of maintaining the BBB between LCH and BO against IS and to further explore the potential mechanism. MATERIALS AND METHODS: After primary mouse brain microvascular endothelial cells (BMECs) were extracted and identified, the duration of oxygen-glucose deprivation (OGD) and the doses of LCH and BO were optimized. Then, the cells were divided into five groups: control, model, LCH, BO, and LCH + BO. Cell viability, injury degree, proliferation and migration were detected by CCK-8, LDH, EdU and wound-healing assays, respectively. Hoechst 33342 staining was adopted to detect the apoptosis rate, and western blotting was employed to observe the expressions of Bax, Bcl-2, caspase-3 and cleaved caspase-3. The TEER value and NaF permeability were measured to assess tight junction (TJ) function, while ZO-1, occludin and claudin-5 were also probed by western blotting. Moreover, the HIF-1α/VEGF pathway was observed to explore the underlying mechanism of BBB maintenance. In vivo, global cerebral ischaemia/reperfusion (GCIR) surgery was performed to establish an IS model. After treatment with LCH (200 mg/kg) and/or BO (160 mg/kg), histopathological structure and BMECs repair were observed by HE staining and immunohistochemistry of vWF. Meanwhile, TJ-associated proteins in vivo were also detected by western blotting. RESULTS: Basically, LCH and BO had different emphases. LCH significantly attenuated the vacuolar structure, nuclear pyknosis and neuronal loss of GCIR mice, while BO focused on promoting BMECs proliferation and angiogenesis and inhibiting the degradation of TJ-associated proteins in vivo after IS. Interestingly, their combination further enhanced these effects. OGD injury markedly reduced the viability, proliferation and migration of primary BMECs; decreased the ratio of Bcl-2/Bax, TEER value, and the expressions of ZO-1, occludin and claudin-5; induced LDH release and apoptosis; and increased the cleaved caspase-3/caspase-3 ratio and NaF permeability. Meanwhile, BO might be the main contributor to the combinative treatment in ameliorating OGD-induced damage of BMECs and degradation of TJ-related proteins, and the potential mechanism might be involved in upregulating the HIF-1α/VEGF signalling pathway. Although LCH showed no obvious improvement, it could enhance the therapeutic effect of BO. Interestingly, their combination even produced some new improvements, including the reduction of cleaved caspase-3 and increase in TEER value, none of which were exhibited in their monotherapies. CONCLUSIONS: LCH and BO exhibited complementary therapeutic features in alleviating cerebral ischaemic injury by inhibiting BMECs apoptosis, maintaining the BBB and attenuating the loss of neurons. LCH preferred to protect ischaemic neurons, while BO played a key role in protecting BMECs, maintaining the BBB and TJs by activating the HIF-1α/VEGF signalling pathway.

Nobiletin inhibits breast cancer cell migration and invasion by suppressing the IL-6-induced ERK-STAT and JNK-c-JUN pathways.

BACKGROUND: Breast cancer is one of the most common cancers in women, affecting more than 2 million women worldwide annually. However, effective treatments for breast cancer are limited. Nobiletin is a flavonoid present in the dried mature pericarp of mandarin orange (Citrus reticulata Blanco), which is used to prepare Citri Renetulatae Pericarpium and can inhibit tumour growth and progression according to modern pharmacological studies. However, whether nobiletin exhibits an antimetastatic role in breast cancer and its potential mechanism need to be further investigated. PURPOSE: This study aims to evaluate the inhibitory effect of nobiletin on breast cancer and to elucidate potential mechanisms against invasion and migration. METHODS: Cell viability was determined by cell counting kit-8 and colony formation assays. Wound healing and Boyden chamber assays detected cancer cell migration and invasion capabilities. Immunoblotting and qPCR were applied to determine the protein and mRNA expression levels of extracellular signal-regulated kinases (ERK) and the c-Jun N-terminal kinase (JNK) signalling pathways. Molecular docking was used to assess the degree of nobiletin binding to phosphatidylinositol 3-kinase (PI3K). Xenografts and liver metastases were constructed in BALB/c nude mice to evaluate the anticancer effect of nobiletin in vivo. H&E staining and immunohistochemistry were used to detect proliferation and the expression of related proteins. RESULTS: Nobiletin induced cell death in a concentration- and time-dependent manner and possessed anti-invasion and anti-migration effects on MCF-7 and T47D cells by suppressing the interleukin-6-induced ERK and JNK signalling pathways. In addition, nobiletin docked with the binding site of PI3K, and the binding score was -8.0 kcal/mol. Furthermore, the inhibition of breast cancer growth and metastasis by nobiletin was demonstrated by constructing xenografts and liver metastases in vivo. CONCLUSION: Nobiletin inhibited liver metastasis of breast cancer by downregulating the ERK-STAT and JNK-c-JUN pathways, and its safety and efficacy were verified, indicating the potential of nobiletin as an anticancer agent.

The mechanism of FZXJJZ decoction suppresses colorectal liver metastasis via the VDR/TGF-ß/Snail1 signaling pathways based on network pharmacology-TCGA data-transcriptomics analysis.

ETHNOPHARMACOLOGICAL RELEVANCE: Fuzheng Xiaojijinzhan (FZXJJZF) decoction is an effective prescription for treating colorectal cancer liver metastasis (LMCRC). AIM OF THE STUDY: To elucidate the pharmacological mechanism of the FZXJJZF decoction therapy on LMCRC. MATERIALS AND METHODS: Firstly, a network pharmacological approach was used to characterize the underlying targets of FZXJJZF on LMCRC. Secondly, LMCRC-related genes are obtained from the public database TCGA, and those genes are further screened and clustered through Mfuzz, an R package tool. Then, targets of FZXJJZF predicted by network pharmacology were overlapped with LMCRC related genes screened by Mfuzz. Meanwhile, FZJZXJF intervened in LMCRC model,epithelial-to-mesenchymal transition (EMT), and migration and invasion of HCT-116 cells. Thirdly, the transcriptomics data of FZJZXJF inhibited HCT-116 cells of EMT cells were overlapped with EMT database data to narrow the possible range of targets. Based on this, the potential targets and signal pathways of FZJZXJF were speculated by combining the transcriptomics data with the targets from network pharmacology-TCGA. Finally, the anti-cancer mechanism of FZXJJZF on LMCRC was verified in vitro by Real-Time PCR and Western Blot in vitro. RESULTS: By network pharmacological analysis, 282 ingredients and 429 potential targets of FZXJJZF were predicted. The 9268 LMCRC-related genes in the TCGA database were classified into 10 clusters by the Mfuzz. The two clustering genes with the most similar clustering trends were overlapped with 429 potential targets, and 32 genes were found, such as CD34, TRPV3, PGR, VDR, etc. In vivo experiments, FZJZXJF inhibited the tumor size in LMCRC models, and the EMT, migration, and invasion of HCT-116 also be inhibited. Intersecting transcriptomics dates with 32 target genes, it is speculated that the VDR-TGF-ß signaling pathway may be an effective mechanism of FZXJJZF. Additionally, it is shown that FZXJJZF up-regulated the expression levels of VDR and E-cadherin and down-regulated the expression levels of TGF-ß and Snail1 in vitro. These results confirmed that FZXJJZF plays an effective role in LMCRC mainly by inhibiting EMT phenotype via the VDR-TGF-ß signaling pathway. CONCLUSIONS: Collectively, this study reveals the anti-LMCRC effect of FZXJJZF and its potential therapeutic mechanism from the perspective of potential targets and potential pathways.

Synergic Effect of Borneol and Ligustrazine on the Neuroprotection in Global Cerebral Ischemia/Reperfusion Injury: A Region-Specificity Study.

The cooperation of ligustrazine (LI) and borneol was proved to be much better than each of them in treating cerebral ischemia. However, the mechanism of their synergic therapy is unclear till now. Moreover, whether their cooperation brought different degrees of protection among different brain regions was also unclear. In the present study, the effects of LI, borneol, and their mixture were observed in global cerebral ischemia-reperfusion (GCIR) injury by detecting microcirculation, expressions of caspase-3 and p53, levels of IL-1ß, IL-6, and TNF-α, and contents of SOD, GSH-Px, and MDA in cortex, hippocampus, hypothalamus, and striatum, respectively. Furthermore, Nissl bodies were scored also. Monotherapy of LI or borneol showed obvious improvements in the four regions, specially in cortex and hippocampus. Interestingly, the cooperation of LI and borneol brought some new improvements, specially in hypothalamus and striatum. Thus, the synergic effect of the two drugs showed region-specificity in GCIR injury except the expressions of caspase-3 and p53.

Cordyceps sinensis Oral Liquid Inhibits Damage Induced by Oxygen and Glucose Deprivation in SH-SY5Y Cells.

CONTEXT: Cordyceps sinensis has been used in traditional Chinese medicine for thousands of years. It has been demonstrated to have a variety of biological activities, and an extract of it has been demonstrated to possess a protective effect in occlusion-induced focal cerebral ischemia of the middle cerebral artery in rats. It could be explored as an agent for treatment of ischemic stroke, and the mechanisms need to be studied further. OBJECTIVE: The study intended to investigate the protective effects of the Cordyceps sinensis oral liquid (CSOL) against damage induced by oxygen and glucose deprivation (OGD) in SH-SY5Y cells. DESIGN • The research team designed an in vitro study. SETTING: The study occurred at the Naval Medical Research Institute in Shanghai, China. INTERVENTION: SH-SY5Y cells were exposed to CSOL in doses of 0.01, 0.03, 0.10, 0.30, and 1.00 mg/mL, creating 5 intervention groups. The OGD condition was induced by transfer of the cells from high-glucose Dulbecco's Modified Eagle's medium (DMEM) in a box gassed with air containing 5% CO2 to glucose-free DMEM in a box gassed with 94% N2, 5% CO2, and 1% O2. Like the cells for the interventions groups, the cells for a model group were cultured with high-glucose DMEM and were transferred to the OGD, but they received no dose of COSL. Cells in a control group were cultured with high-glucose DMEM, were not transferred to the OGD condition, and did not receive any dose of COSL. OUTCOME MEASURES: Cell viability was assayed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. The apoptosis and the mitochondrial membrane potential (MMP) were detected by flow cytometry, and the protein expression of caspase-3 was observed by western blot. RESULTS: After exposure to OGD, the cell viability of cells treated with 0.01, 0.03, 0.10, 0.30, and 1.00 mg/mL of CSOL increased in a dose-effect relationship. Compared with the cells in the model group, the treatment of CSOL at all the experimental concentrations significantly inhibited both the cell apoptosis (P < .01) and the capase-3 activation (P < .01). The MMP dissipation in the cells of the model group increased significantly compared with those of the control group (P < .01). The treatment with all doses of CSOL significantly inhibited the MMP dissipation (P < .01). CONCLUSIONS: CSOL protects against the damage induced by OGD through inhibiting the mitochondrial apoptosis pathway in SH-SY5Y cells.

Attenuation of stress-induced gastrointestinal motility disorder by gentiopicroside, from Gentiana macrophylla Pall.

AIM: The current study was designed to explore the mechanism of the prokinetic activity of Gentiopicroside (Ge), from Gentiana macrophylla Pall which is widely used to strengthen gastric motility in clinic. METHODS: Gastrointestinal motility disorder rats were induced by stress stimulation and the rats were treated with Ge. The functions of gastric emptying and intestinal propelling were measured after blood was obtained to assay the levels of plasmatic motilin (MTL), vasoactive intestinal peptide (VIP), somatostatin (SST), gastrin (GAS), neurotensin (NT) and substance of P (SP). The expressions of MTL receptor (MTLR), VIP receptor 2 (VIPR2) and SST receptor 2 (SSTR2) were measured also. In addition, an isolated guinea pig ileum was applied to evaluate the influences of Ge on M-R, H1-R, 5-HT4-R and D-R in vitro. RESULTS: Ge increased gastric emptying and intestinal propelling obviously. It also decreased the level of SST and increased GAS in plasma significantly. Moreover, it promoted the expressions of MTLR in gastric antrum, duodenum, jejunum and ileum, and restrained the expression of VIPR2 in duodenum. Piboserod and loratadine had no obvious restrain to Ge' exciting ileum effect and Ge also didn't affect dopamine paralyzing ileum. However, Ge failed to improve the hypofunction of guinea pigs ileums pre-treated with atropine sulfate. CONCLUSION: The mechanisms of Ge' prokinetic effect were associated with modulating the levels of SST and GAS in plasma, raising the expressions of MTLR in gastric antrum, duodenum, ileum and jejunum, reducing the expression of VIPR2 in duodenum and activating M-R.

[Effect of electroacupuncture intervention on expression of ATP-binding cassette protein and gene in the cerebral cortex of depression rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on the expression of blood-brain barrier (BBB)-related functional protein, ATP-binding cassette protein and gene in the cerebral cortex of depression rats. METHODS: Forty SD rats were equally and randomly divided into control group, EA group, depressive (model) group and depression + EA group. The depression model was established by chronic unpredictable mild stress. EA (2 Hz, 1 mA) was applied to "Baihui"(GV 20) and "Yamen"(GV 15) for 20 min, once daily for 14 days. The expression levels of multidrug resistance (Mdr) 1a, Mdr 1b, multidrug resistance-associated protein (Mrp) 1, Mrp 4, Mrp 5, and breast cancer resistance protein (Bcrp) gene and proteins in the cerebral cortex tissue were detected by RT-PCR semi-quantitative analysis and Western blot, respectively. RESULTS: In comparison with the control group, no obvious changes were found in the expression levels of cerebral Mdr la, Mdr 1b, Mrp 1, Mrp 4, Mrp 5 and Bcrp genes and proteins in the model group (P > 0.05), and only cerebral Mdr 1a gene and protein expression levels in the EA group were evidently down-regulated (P < 0.01, P < 0.05). Compared with the model group, only cerebral Mdr la gene and protein expression levels were obviously decreased in the depression+ EA group (P < 0.01, P < 0.05). No significant changes were found in the Mdr 1b, Mrp 1, Mrp 4, Mrp 5 and Bcrp genes and proteins after EA intervention in depression rats (P > 0.05). CONCLUSION: EA of GV 20 and GV 15 can obviously down-regulate the expression of cerebral Mdr 1a in depression rats which might be involved in the opening activity of blood-brain barrier.

Effects of borneol on the pharmacokinetics of geniposide in cortex, hippocampus, hypothalamus and striatum of conscious rat by simultaneous brain microdialysis coupled with UPLC-MS.

It has been verified that borneol could promote the accumulation of other drugs in the whole brain. In this study, a microdialysis sampling system coupled with UPLC-MS was developed to evaluate the delivery of geniposide to four brain regions (cortex, hippocampus, hypothalamus and striatum) of conscious rats in the absence/presence of borneol: rats were administrated with geniposide alone (300mg/kg, iv) or administrated with both geniposide and borneol (0.2g/kg, ig). The dialysate collected from specific brain area was analyzed by a UPLC-MS system: separated on a BEH C18 column (50mm×2.1mm id, 1.7µm) within 1.5min, and detected in positive ion electrospray mode. The calibration curve was in good linearity over the concentration range of 0.009-90µg/mL. The inter- and intra-day accuracies were within ±10%, and the precisions were within 9.13%. The established method was applied to study the brain pharmacokinetics of geniposide and the results demonstrated that borneol markedly facilitated the delivery of geniposide to hippocampus and hypothalamus, but slightly hampered its delivery in cortex.

The mechanism of the opening of the blood-brain barrier by borneol: a pharmacodynamics and pharmacokinetics combination study.

ETHNOPHARMACOLOGICAL RELEVANCE: Borneol is widely used in traditional Chinese medicine to facilitate the distribution of central nervous system (CNS) drugs in brain due to its ability to open blood-brain barrier (BBB), however, the underlying mechanism is still unclear. In this study, the effect of borneol on different brain regions were investigated to explore the mechanism. MATERIALS AND METHODS: After oral administration of borneol (0.1, 0.2 g/kg) for seven consecutive days, SD rats were injected with Rh123 (1.0 mg/kg). The concentrations of Rh123 were detected in four brain regions of cortex, hippocampus, hypothalamus and striatum by a small animal vivo imaging system and a fluorescence microplate reader respectively. The ultrastructures of BBB were examined. Moreover, the expressions of the four transporters of ATP-binding cassette (ABC) family, multidrug resistance 1a (Mdr1a), multidrug resistance 1b (Mdr1b), multidrug resistance protein 1 (Mrp1), Mrp4, Mrp5 and breast cancer resistance protein (Bcrp) in the four brain regions were analyzed. Finally, the deliveries of borneol in the plasma and the four brain regions were examined by a pharmacokinetics study. RESULTS: Administration of 0.2 g/kg borneol produced loose structure in the tight junction and void structure between the endothelial cell and mesangial cell. Borneol at 0.1 g/kg and 0.2 g/kg increased the delivery of Rh123 in hippocampus and hypothalamus obviously. Permeability index followed a similar trend. Protein expression assays showed that borneol decreased the expression of Mdr1 and Mrp1 in hippocampus and hypothalamus. Further RT-PCR study showed that borneol decreased the expressions of both Mdr1a and Mdr1b in hippocampus and hypothalamus. The pharmacokinetics study demonstrated that the delivery of borneol in cortex was the most and that in striatum the least, with the deliveries of borneol in hippocampus and hypothalamus in between. CONCLUSIONS: Borneol showed tissue specific BBB-opening effect, which was associated with its regulation of the ultrastructure of brain tissues and the expressions of Mdr1a, Mdr1b and Mrp1. The present study indicated that borneol should be used in concert with drugs targeting hippocampus or hypothalamus to exert its synergistic effect to the maximum.

Influence of cyclovirobuxine D on intracellular [Ca(2+)] regulation and the expression of the calcium cycling proteins in rat myocytes.

Our previous studies have shown that cyclovirobuxine D (CVB-D) ameliorated the cardiac function in heart failure rats. Considering the relationship between cardiac function and [Ca(2+)]i, and the role of calcium cycling on regulating [Ca(2+)]i, the present study was designed to evaluate the influence of CVB-D on the calcium transient of myocytes from neonatal rats and adult heart failure (HF) rats. The expression of calcium cycling proteins, including L-type calcium channel (LTCC), ryanodine receptor 2 (RYR2), sarcoplasmic reticulum calcium ATPase 2a (SERCA2a) and sodium-calcium exchanger (NCX), were investigated to explore the underlying mechanism. CVB-D increased the intensity of calcium transient, accelerated the process of calcium transient and attenuation in the neonatal and adult myocytes. Furthermore, CVB-D shortened T(peak) and T(attenuation) in the adult myocytes and slowed down the heart rate of neonatal myocytes. Besides, CVB-D increased the expression of RYR2 and SERCA2a, decreased the expression of NCX, but showed no significant effect on LTCC. Thus, it was concluded that CVB-D increased the release and uptake of Ca(2+) in systolic and diastolic period, respectively. CVB-D might not only facilitate the utilization of intracellular Ca(2+), but also prevent the loss of Ca(2+).

Qualitative and quantitative analysis of the major constituents in Chinese medicinal preparation Guan-Xin-Ning injection by HPLC-DAD-ESI-MS(n).

Guan-Xin-Ning (GXN) injection, a traditional Chinese medicinal preparation consisting of Radix Salvia miltiorrhiza and Rhizoma Ligusticum chuanxiong, has been used to treat coronary heart disease and angina pectoris in China for decades. In this paper, a HPLC/DAD/ESI-MS(n) method was successfully developed for qualitative and quantitative analysis of the active components in GXN injection for the first time. 28 compounds were identified by comparison of their retention times and MS spectra (HPLC/DAD/ESI-MS(n)) with those elucidated standards or recorded literature. 19 of them (danshensu, furoic acid, 3-O-caffeoylquinic acid, protocatechuic aldehyde, p-hydroxybenzoic acid, chlorogenic acid, caffeic acid, 4-O-caffeoylquinic acid, vanillin, 1,3-dicaffeoylquinic acid, 4-hydroxycinnamic acid, ferulic acid, senkyunolide I, senkyunolide H, isosalvianolic acid A, rosmarinic acid, salvianolic acid B, salvianolic acid A and isosalvianolic acid C) were simultaneously determined by HPLC-DAD quantitatively. The analytical method was validated and successfully applied for simultaneous determination of major components in GXN injections from seven different production batches, indicating that the proposed approach was applicable for the routine analysis and quality control of GXN injection.

Acute morphine administration and withdrawal from chronic morphine increase afterdepolarization amplitude in rat supraoptic nucleus neurons in hypothalamic explants.

Supraoptic nucleus (SON) neurons secrete either oxytocin or vasopressin into the bloodstream from their axon terminals in the posterior pituitary gland. SON neurons are powerfully inhibited by the classical µ-opioid receptor agonist, morphine. Oxytocin neurons develop morphine dependence when chronically exposed to this opiate, and undergo robust withdrawal excitation when morphine is subsequently acutely antagonized by naloxone. Morphine withdrawal excitation is evident as an increased firing rate and is associated with an increased post-spike excitability that is consistent with the expression of an enhanced post-spike afterdepolarization (ADP) during withdrawal. Here, we used sharp electrode recording from SON neurons in hypothalamic explants from morphine naïve and morphine treated rats to determine the effects of morphine on the ADP, and to test the hypothesis that morphine withdrawal increases ADP amplitude in SON neurons. Acute morphine administration (0.05-5.0 µM) caused a dose-dependent hyperpolarization of SON neurons that was reversed by concomitant administration of 10 µM naloxone, or by washout of morphine; counter-intuitively, acute exposure to 5 µM morphine increased ADP amplitude by 78 ± 11% (mean ± SEM). Naloxone-precipitated morphine withdrawal did not alter baseline membrane potential in SON neurons from morphine treated rats, but increased ADP amplitude by 48 ± 11%; this represents a hyper-activation of ADPs because the basal amplitude of the ADP was similar in SON neurons recorded from explants prepared from morphine naïve and morphine treated rats. Hence, an enhanced ADP might contribute to morphine withdrawal excitation of oxytocin neurons.

[Effects of CVB-D on hemodynamic in anaesthetized dogs in vivo].

OBJECTIVE: To investigate hemodynamic in anaesthetized dogs after the intravenous injection of cyclovirobuxine D (CVB-D). METHODS: The hemodynamic of anaesthetized dogs were observed after intravenous injection of CVB-D at doses of 0.1, 0.2, 0.4 mg/kg. RESULTS: CVB-D of 0.2 and 0.4 mg/kg could decrease HR, TPVR and increase CBF. In addition, CVB-D of 0.4 mg/kg could increase SAP and SV. Yet MAP and CO of dogs showed no remarkable changes with the treatment of CVB-D. CONCLUSION: CVB-D has effect of improving cardiac function, which may be the mechanism of anti-myocardial ischemia effect of CVB-D.

[Transformation of astragaloside IV in bidirectional solid fermenting of Astragalus membranaceus].

OBJECTIVE: To study the transformation of Astragaloside IV in bidirectional solid fermenting of Astragalus membranaceus var. mongholicus. METHODS: The chemical components of pre and post-fermenting were comparatively analyzed by techniques of HPLC, column chromatography and spectral identification. Antioxidant effect of transformational product on vascular endothelial cells was researched. RESULTS: In fermenting of Astragalus membranaceus, Astragaloside IV was transformed into 6-O-beta-D-glucoside cycloastragenol, which had significant antioxidant effect. CONCLUSION: Biotransformation of Astragaloside IV in Astragalus membranaceus was occurred after bidirectional solid fermenting with Ganoderma lucidum, and the biotransformation could enhance effect.

[The hepatoprotective effect of aqueous extracts from Ficus hirta on N, N-dimethylformamide induced acute liver injury in mice].

OBJECTIVE: To investigate the effect of aqueous extract from Ficus hirta on N, N-Dimethylformamide (DMF) induced liver injury in mice. METHODS: C57BL/6 mice and ICR mice were randomly divided into 5 groups: negative control group, positive control group and three treated groups respectively. Treated groups were administered orally with 100, 200, 300 g/kg Ficus hirta aqueous extract per day respectively for 5 days. On the 4th day, 2. 3 g/kg DMF was given by intraperitoneal injection to all C57BL/6 mice except negative control group, while for ICR, DMF was administration at a 2.75 g/kg dose. 48h after DMF injection, serum samples were collected to determine the activities of ALT, AST and LDH and the pathological changes of liver tissue were analyzed under microscope. RESULTS: Compared with the positive control, the activities of ALT, AST and LDH were significantly reduced and the liver injury obviously attenuated in treated groups. CONCLUSION: The aqueous extract of Ficus hirta has an obvious protective effect against DMF-induced acute liver injury in mice.

[Effect of roots of Ficus hirta on cocaine-induced hepatotoxicity and active components].

OBJECTIVE: To investigate the protective effect of the roots of F. hirta against the cocaine-induced hepatotoxicity and it's active components. METHOD: Cocaine hydrochloride was subcutaneously injected to make male ICR mice liver wounded. Male ICR mice were randomly ig administered with the F. hirta decoction. The dose groups are 100, 200, 300 g x kg(-1) herb materials per body weight. Cocaine hydrochloride was subcutaneously injected into the mice after the administration. The serum ALT, AST activity and the activity of CAT in liver homogenate were assayed, and liver change of pathomorphism was evaluated to prove the effect of the F. hirta decoction on cocaine-induced hepatotoxicity. And the activity of psoralean which was separated from the F. hirta decoction by bioassay-guided fractionation, was proofed in the same method. RESULT: We find that the F. hirta decoction shows a distinct effect on reducing serum transferase. The serum transferase and the content CAT in liver homogenate were dose-related reduced, and the histopathological examination found a significantly change of the liver tissues. And the psoralean, qua the mainly component, shows the same effect. CONCLUSION: F. hirta has the protective effect against the cocaine-induced hepatotoxicity. Psoralean is the basis.