Chongqing Engineering Research Center for Pharmacological Evaluation

Chongqing, China

Chongqing Engineering Research Center for Pharmacological Evaluation

Chongqing, China
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Liu Y.,Southwest University | Liu Y.,Chongqing Engineering Research Center for Pharmacological Evaluation | Tang Q.,Southwest University | Tang Q.,Chongqing Engineering Research Center for Pharmacological Evaluation | And 7 more authors.
International Journal of Biological Sciences | Year: 2017

Catalpol and puerarin are two monomers of Rehmannia glutinosa and Lobed Kudzuvine Root, which are two herbs commonly used together in ancient prescriptions of traditional Chinese medicine for cerebral ischemia. Our previous study shows that the lyophilized powder of the two monomers improved the outcome of cerebral ischemia excellently in rodents. However, if it protects vessels from ischemia is unknown. The present research studied the protection of lyophilized powder of catalpol and puerarin (CP) on endothelial cells and the relative mechanism in vivo and in vitro. Middle cerebral artery occlusion (MCAO) rats were used to study the improvement of CP on neurological deficiency, regional cerebral blood flow (rCBF), and infarct volume. The morphology of vessels and the apoptosis of brain vascular endothelial cells (BVECs) were observed and detected by immunohistochemistry approaches. To study how CP protected primary BVECs (pBVECs) from ischemic penumbra, oxygen glucose deprivation (OGD)-damaged pBVECs were cultured in the condition of insufficient nutrition and low oxygen which recapitulate the low perfusion of ischemic penumbra. Using the cell model, the mechanism by which CP protected pBVECs was studied by shRNA and pathway inhibitors. CP at the dose of 65.4 mg/kg increased regional cerebral blood flow (rCBF), reduced infarct volume, protected vessel integrity and inhibited endothelial cell apoptosis in vivo. But it only improved rCBF, vessel integrity and BVECs apoptosis at the dose of 32.7 mg/kg. In vitro, the protection of CP on pBVECs was proved to be ERK/HIF-1a- and PI3K/AKT/mTOR/HIF-1a-dependent. This study indicates a possibility of CP being a new drug for cerebral ischemia. Besides, this research provides an alternative cell model for penumbra ECs study. © Ivyspring International Publisher.


Zhu H.,Southwest University | Zhu H.,Chongqing Engineering Research Center for Pharmacological Evaluation | Wang Y.,Southwest University | Wang Y.,Chongqing Engineering Research Center for Pharmacological Evaluation | And 10 more authors.
Chinese Medicine (United Kingdom) | Year: 2016

Background: Diabetes, associated with hyperlipidemia and oxidative stress, would lead to an increased production of reactive oxygen species. Rehmannia glutinosa (Di Huang) is widely used to nourish yin, invigorate the kidney (shen), and treat xiao ke (a diabetes-like syndrome in Chinese medicine). This study aims to investigate the antidiabetic and antioxidant effects of catalpol from R. glutinosa on rat diabetes induced by streptozotocin (STZ) and high-fat, high-sugar feed. Methods: Rats (eight rats in each group at least) were induced diabetes by an initial high-fat high-sugar feed for 3weeks, followed by an intraperitoneal injection of STZ (30mg/kg) for 3days, and rats were fasted overnight before treatments. Catalpol at a dose of 0, 5, 10, 20 or 50mg/kg was administrated through bolus intravenous injection to the experimental rats to find the most effective anti-hyperglycemic dose of catalpol to further study body weight loss, water intake, and food intake. The most effective catalpol dose was given to the diabetic model rats with hyperlipidemia, and the levels of blood sugar, plasma total cholesterol (TC), triglyceride (TG), and high-density lipoprotein cholesterol (HDL-C) were measured after catalpol administration once a day for 2weeks. An oral glucose challenge test (OGCT) was performed after above experiments in which the most effective dose of catalpol has been determined. Levels of glutathione peroxidase (GSH-PX), catalase (CAT), superoxide dismutase (SOD), and malondialdehyde (MDA) were measured by corresponding reagent kits and morphological changes of the pancreas were observed with histopathological examination using H&E stain. Results: Catalpol at a dose of 50mg/kg ameliorated body weight loss and increased water and food intake. Catalpol also attenuated the increase of plasma TC (P=0.0067) and TG (P=0.0084) and increased HDL-C (P=0.0336). The OGCT revealed that catalpol reduced the increase of plasma glucose. The activities of antioxidative enzymes (SOD, P=0.0037; GSH-PX, P=0.0066; CAT, P=0.005) were enhanced and MDA was reduced (P=0.003). Furthermore, catalpol reduced the morphological impairment of the pancreas. Conclusion: Catalpol protected against STZ-induced diabetes with high-fat and high-sugar feed with ameliorated structural impairment of the pancreas and restored balance between oxidative enzymes and antioxidative enzymes. © 2016 Zhu et al.


Tang D.-D.,Southwest University | Li N.,Southwest University | Wang L.-W.,Southwest University | Zhang J.-F.,Southwest University | And 2 more authors.
Zhongguo Zhongyao Zazhi | Year: 2014

To study the interaction of drugs of different properties, namely puerarin, borneol and catalpol iu the process of inclusion, in order to explore the inclusion regilarity of inulti-coiujxnient and multi-property traditional Chinese medicine compound inclusions. With HP-β-CD as the inclusion material, the freeze-drying method was used to prepare the inclusion. The inclusion between puerarin, borneol and catalpol was tested by i easuring the inclusion concentration, DSC and X-ray diffraction. According to the findings, when insoluble drugs puerarin and lx>rneol were included simultaneously, and puerarin was ovenlosed, puerarin included was almost equal to puerarin included, and bonieol was not included. When puerarin was under-dosed, and HP-β-CD was overdosed, borneol was included, and the simultaneous inclusion was lower than the separate inclusion of borneol. When water-soluble drug catalpol was jointly included with puerarin or borneol, the simultaneous inclusion was almost the same with their separate inclusion, without characteristic peak of catalpol iu DSC and X-ray diffraction patterns. There is a competition iu the simultaneous inclusion between water-soluble drugs puerarin and borneol and a stronger competition iu puerarin. The water-soluble drug catalpol could lx included with HP-β-CD with no impact on the inclusion of puerarin or borneol.


Xue Q.,Southwest University | Liu Y.,Southwest University | He R.,Southwest University | Yang S.,Southwest University | And 5 more authors.
International Journal of Biological Sciences | Year: 2016

Hunting for an effective medicine for brain stroke has been a medical task in neuroscience for decades. The present research showed that the lyophilized Powder of Catalpol and Puerarin (C-P) in all the tested doses (65.4 mg/kg, 32.7 mg/kg, 16.4 mg/kg) significantly reduced the neurological deficiency, infarct volume and apoptotic cells in ischemic/reperfusion (I/R) rats. It also promoted astrocyte processes and prolonged neuron axons in infarct area. Further, it decreased MDA, NO, NF-κB/p65, TNF-α, IL-1β and IL-6 and enhanced the EPOR and GAF-43. 65.4 mg/kg and 32.7 mg/kg C-P could up-regulated EPO and VEGF significantly. In vitro, 49 μg/mL and 24.5 μg/mL C-P decreased the leakage of sodium fluorescein and increased the activity of γ-GTP. Additionally, it increased SOD and decreased MDA, NO, and LDH and decreased NF-κB/p65, TNF-α, IL-1β and IL-6 and unregulated EPO, EPOR, VEGF, and GAP-43. Only the dose of 49 μg/mL increased TEER and Claudin-5 and turned the typically damaged morphologies of neurons, astrocytes and endothelium into a favorable trend. These data imply that C-P improved the recovery of neurological deficiency in motor, sense, balance and reflex, and protected the whole NVU by anti-oxidative stress, anti-inflammation and up-regulating some protective factors. This research provides a candidate medicine for brain stroke and, at the same time, a pattern for drug study targeting NVU in vitro. © Ivyspring International Publisher.


Xue Q.,Southwest University | Xue Q.,Chongqing Engineering Research Center for Pharmacological Evaluation | Liu Y.,Southwest University | Liu Y.,Chongqing Engineering Research Center for Pharmacological Evaluation | And 10 more authors.
International Journal of Biological Sciences | Year: 2013

A novel triple cell neurovascular unit (NVU) model co-culturing with neurons, brain microvascular endothelial cells (BMECs) and astrocytes was established in this study for investigating the cerebral diseases and screening the candidates of therapeutic drug. We have first performed the cell identification and morphological characterization, analyzed the specific protein expression and determined the blood-brain barrier (BBB) function of the co-culture model under normal condition. Then, we further determined the BBB function, inflammation, cell injury and the variation of neuroprotective factor in this model after anoxia-reoxygenation. The results suggest that this model exhibited a better BBB function and significantly increased expression of P-glycoprotein (Pg-P) and ZO-1 compared with BMECs only or co-culture with astrocytes or neurons. After anoxia-reoxygenation, the pathological changes of this model were basically resemblance to the pathological changes of brain cells and BBB in vivo. And nimodipine, an antagonist of calcium, could reverse those changes as well. According to our observations, we deduce that this triple cell co-culture model exhibits the basic structure, function and cell-cell interaction of NVU, which may offer a more proper in vitro system of NVU for the further investigation of cerebral diseases and drug screening. © Ivyspring International Publisher.


Li X.,Southwest University | Li X.,Chongqing Engineering Research Center for Pharmacological Evaluation | Chen Y.,Southwest University | Chen Y.,Chongqing Engineering Research Center for Pharmacological Evaluation | And 7 more authors.
Experimental Gerontology | Year: 2016

Brain Erythropoietin (EPO), an important neurotrophic factor and neuroprotective factor, was found to be associated with aging. Studies found EPO expression was significantly decreased in the hippocampus of aging rat compared with that of the youth. But mechanisms of the decline of the brain EPO during aging remain unclear. The present study utilized a D-galactose (D-gal)-induced aging model in which the inducement of aging was mainly oxidative injury, to explore underlying mechanisms for the decline of brain EPO in aging rats. D-gal-induced aging rats (2 months) were simulated by subcutaneously injecting with D-gal at doses of 50 mg·kg− 1, 150 mg·kg− 1 and 250 mg·kg− 1 daily for 8 weeks while the control group received vehicle only. These groups were all compared with the aging rats (24 months) which had received no other treatment. The cognitive impairment was assessed using Morris water maze (MWM) in the prepared models, and the amount of β-galactosidase, the lipid peroxidation product malondialdehyde (MDA) level and the superoxide dismutase (SOD) activity in the hippocampus was examined by assay kits. The levels of EPO, EPOR, p-JAK2 and hypoxia-inducible factor-2α (HIF-2α) in the hippocampus were detected by western blot. Additionally, the correlation coefficient between EPO/EPOR expression and MDA level was analyzed. The MWM test showed that compared to control group, the escape latency was significantly extended and the times of crossing the platform was decreased at the doses of 150 mg·kg− 1 and 250 mg·kg− 1 (p < 0.05). Also, the amount of β-galactosidase and the MDA level in the hippocampus were significantly increased but the SOD activity was significantly decreased (p < 0.05, 0.01 and 0.01, respectively). Similar to aging rats, the expressions of EPO, EPOR, p-JAK2, and HIF-2αin the brain of D-gal-treated rats were significantly decreased (p < 0.05) at 150 mg·kg− 1 and 250 mg·kg− 1. Interestingly, negative correlations were found between EPOR (r = − 0.699, p < 0.01), EPO (r = − 0.701, p < 0.01) and the MDA level. These results indicated that aging could result in the decline of EPO in the hippocampus and oxidative stress might be the main reason for the decline of brain EPO in aging rats, involved with the decrease of HIF-2α stability. © 2016 Elsevier Inc.


Wan D.,Chongqing Medical University | Xue L.,Chongqing Chemical Industry Vocational College | Zhu H.,Southwest University | Zhu H.,Chongqing Engineering Research Center for Pharmacological Evaluation | Luo Y.,Chongqing Medical University
Evidence-based Complementary and Alternative Medicine | Year: 2013

To investigate the role and mechanism of catalpol on neuroprotective effects and memory enhancing effects simultaneously, neuroprotective effects of catalpol were assessed by neurological deficits score, TTC staining, and cerebral blood flow detecting. Morris water maze was employed to investigate its effects on learning and memory and then clarify its possible mechanisms relating the central cholinergic system and BDNF. Edaravone and oxiracetam were used for positive control drugs based on its different action. Results showed that catalpol and edaravone significantly facilitated neurological function recovery, reduced infarction volume, and increased cerebral blood flow in stroke mice. Catalpol and oxiracetam decreased the escape latency significantly and increased the numbers of crossing platform obviously. The levels of ACh, ChAT, and BDNF in catalpol group were increased in a dose-dependent manner, and AChE declined with a U-shaped dose-response curve. Moreover, the levels of muscarinic AChR subtypes M1 and M2 in hippocampus were considerably raised by catalpol. These results demonstrated that catalpol may be useful for neuroprotection and memory enhancement, and the mechanism may be related to the central cholinergic system. © 2013 Dong Wan et al.


Tang D.-D.,Southwest University | Zhang J.,Southwest University | Wang J.-R.,Southwest University | Zhang J.-F.,Southwest University | And 2 more authors.
Zhongguo Zhongyao Zazhi | Year: 2016

To investigate the effect of borneol on the oral absorption and penetration into brain of puerarin and catalpol from cell level and animal level, and screen the concentration of borneol that is suitable for Zige compound oral preparation. Bhxxl-brain barrier ( BBB) model was established by co-culture of primary brain miemvessel endothelial eells(BMEC) and astrocytes( As) in rats, and it was used to investigate the effect of borneol (concentration from 6. 25 to 100 mg • L-1) on the transport of puerarin and catalpol. The pharmacokinetics of puerarin and catalpol in plasma and brain of rats were compared after intragastric administration of borneol solution (0, 25, 50 and 100 mg • kg -1) immediately followed by puerarin( 200 mg • kg -1) and catalpol(45 mg • kg -1) nanocrystal suspension. Barrier function was basically formed after co-eulturing of brain microvascular endothelial cells and astrocytes for 7 d. The permeability of puerarin and catalpol across blood-brai barrier was increased significantly (P < 0.05) and transendothelial electrical resistance(TEER) values at 2 h were decreased signifieantly(P <0. 01) when the concentration of borneol was between 12. 5 to 100 mg • L -1 as compared with the control group. Borneol at the dose of 50 mg • kg -1 and 100 mg • kg -1 could significantly increase the oral ab-sorption of puerarin(P <0. 05) , but there was no obvious ettect tor catalpol. AULbrain/ AUCblood for puerarin was highest with borneol at dose of 100 mg • kg -1 (P <0.05) , while AULbrain/ AUCblood for catalpol was highest with borneol at dose of 50 mg • kg -1 (P < 0.05). AUC brain was highest at 100 mg • kg -1 for puerarin(P < 0.05) ; while for catapol, it was highest at 50 mg • kg -1 , but it was not significantly different from 100 mg - kg -1. In conclusion, borneol could increase the amount of puerarin and catalpol in brain after oral ad- ministration and the optimized dose shall be 100 mg • kg -1.


Liu Y.,Southwest University | Liu Y.,Chongqing Engineering Research Center for Pharmacological Evaluation | Xue Q.,Southwest University | Xue Q.,Chongqing Engineering Research Center for Pharmacological Evaluation | And 10 more authors.
International Journal of Biological Sciences | Year: 2014

Catalpol and puerarin are active ingredients isolated from Rehmannia glutinosa Libosch and Radix Puerariae, respectively. They are popular in research for their poly-pharmacological effects. This research focused on effect of anti-stroke by lyophilized powder of catalpol and puerarin (C-P) and potential mechanisms. At the beginning of research, C-P was identified and analyzed by HPLC. Neurological function was evaluated by Longa score, neurological complex function score and beam balance score after permanent middle cerebral artery occlusion (PMCAO) in mice. Infarct volume and water content were evaluated after treatment of C-P. Anti-oxidative stress, an-ti-apoptosis, angiogenesis and neurogenesis were investigated by ELISA, WB and immunohisto-chemical stain respectively. With treatment of C-P, neurological deficiency of PMCAO mice was ameliorated. Morphologically, infarct volume and water content in ischemic hemisphere were significantly reduced by C-P. In vivo and in vitro, oxidative stress injury was extenuated by C-P. Meanwhile, Caspase-3 was down-regulated and Bxl-2 was up-regulated by C-P in vivo. In addition, C-P enhanced angiogenesis around the infarct of cortex and neurogenesis in the Hippocampal Dentate Gyrus (DG). Hence, C-P ameliorated stroke-induced neurological deficiency through its multiple neuroprotections. What's more, this article provides us a novel formula of active ingredients for stroke. © Ivyspring International Publisher.


Liu Y.,Southwest University | Liu Y.,Chongqing Engineering Research Center for Pharmacological Evaluation | Xue Q.,Southwest University | Xue Q.,Chongqing Engineering Research Center for Pharmacological Evaluation | And 14 more authors.
Microvascular Research | Year: 2013

Brain microvascular endothelial cells (BMECs), a main component of the blood-brain barrier, play a critical role in the pathogenesis of many brain diseases. The primary culture of BMECs has been used in various models for studying cerebrovascular diseases in vitro. However, there are still several problems existing in the isolation and cultivation of primary rat BMECs, such as low yield, contamination with other cell types, and requirement of a large number of animals and expensive growth factor. In this study, we describe a simple, economical (without any growth factor) and repeatable method to obtain endothelial cells with high purity (>99%) and yield (about 2.2×107 per rat) from cerebral cortexes of neonatal rat, mainly from gray matter. In vitro examinations determined that the isolated cells expressed typical phenotypic markers of differentiated brain endothelium such as multiple drug resistant protein, von Willebrand factor, platelet endothelial cell adhesion molecule 1 (PECAM-1/CD31), and intercellular adhesion molecule (ICAM). These cells also possessed morphological and ultra-structural characteristics that were observed by phase contrast microscope and electric microscope. Then GFAP and α-SMA were used, respectively, to identify astrocyte and pericyte which were potential to contaminate primary culturing of BMECs. And specific reaction of endothelial cells to external stimulation was tested by culture with TNF-α for 24h. All these results of our experiments supply that our protocol provides an effective and reliable method to obtain high purity and yield of rat BMECs and offers a useful tool for studying cellular physiology, cerebrovascular diseases, brain tumors, blood-brain barrier and neurovascular units, etc. © 2013 Elsevier Inc.

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