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Yang G.,Key Laboratory Of Xinan Medicine | Yang G.,Anhui University | Yang G.,Anhui Academy of Chinese Medicine | Qian C.,Key Laboratory Of Xinan Medicine | And 17 more authors.
Cellular and Molecular Neurobiology | Year: 2016

Tetramethylpyrazine (TMP, also known as Ligustrazine), which is isolated from Chinese Herb Medicine Ligustium wollichii Franchat (Chuan Xiong), has been widely used in China for the treatment of ischemic stroke by Chinese herbalists. Brain microvascular endothelial cells (BMECs) are the integral parts of the blood–brain barrier (BBB), protecting BMECs against oxygen-glucose deprivation (OGD) which is important for the treatment of ischemic stroke. Here, we investigated the protective mechanisms of TMP, focusing on OGD-injured BMECs and the Rho/Rho-kinase (Rho-associated kinases, ROCK) signaling pathway. The model of OGD-injured BMECs was established in this study. BMECs were identified by von Willebrand factor III staining and exposed to fasudil, or TMP at different concentrations (14.3, 28.6, 57.3 µM) for 2 h before 24 h of OGD injury. The effect of each treatment was examined by cell viability assays, measurement of intracellular reactive oxygen species (ROS), and transendothelial electric resistance and western blot analysis (caspase-3, endothelial nitric oxide synthase (eNOS), RhoA, Rac1). Our results show that TMP significantly attenuated apoptosis and the permeability of BMECs induced by OGD. In addition, TMP could notably down-regulate the characteristic proteins in Rho/ROCK signaling pathway such as RhoA and Rac1, which triggered abnormal changes of eNOS and ROS, respectively. Altogether, our results show that TMP has a strong protective effect against OGD-induced BMECs injury and suggest that the mechanism might be related to the inhibition of the Rho/ROCK signaling pathway. © 2016 Springer Science+Business Media New York

Wang T.-M.,Anhui University | Wang T.-M.,Anhui Academy of Chinese Medicine | Yan Y.-Y.,Anhui University | Yan Y.-Y.,Anhui Academy of Chinese Medicine | And 8 more authors.
Chinese Traditional and Herbal Drugs | Year: 2014

Objective: To observe the effects of ethyl acetate extract from Huanglian Jiedu Decoction (EAHJD) on the virulence factors of Candida albicans. Methods: Egg-yolk medium, milk-plate medium, and olive oil emulsification were used respectively to test the activities of phospholipase (PL), aspartic protease (Sap), and lipase (Lip) of C. albicans. The water-hydrocarbon two-phase assay was applied to measure the cell surface hydrophobicity (CSH) of C. albicans. qRT-PCR was adopted to observe the expression of virulence factors related genes. Results: EAHJD had no effect on the activity of PL. EAHJD (1 250 μg/mL) could significantly inhibit the activity of PL and Lip better than that by 312 μg/mL EAHJD. EAHJD could reduce CSH of C. albicans in a dose-independent manner and CSH1 was down-regulated by 7.69, 3.57, and 2.95 folds by 1 250, 312, and 78 μg/mL EAHJD, respectively. The expression of secretory enzyme related genes displayed different changing folds: PLC1, Sap2, Sap3, Sap9, Lip3, Lip4, and Lip6 were down-regulated; PLB1, PLC2, Sap1, Sap10, and Lip5 had no distinct change treated by EAHJD. Conclusion: EAHJD could inhibit the activities of virulence factors of C. albicans. ©, 2014, Editorial Office of Chinese Traditional and Herbal Drugs. All right reserved.

Xie D.,Anhui University | Xie D.,Anhui Academy of Chinese Medicine | Zhang E.-L.,Anhui University | Zhang E.-L.,Anhui Academy of Chinese Medicine | And 5 more authors.
Yaoxue Xuebao | Year: 2015

Fifteen novel ligustrazine-tetrahydroisoquinoline derivatives were designed and synthesized according to the association principle of pharmaceutical chemistry. The structures were identified by IR, NMR and ESI-MS. The inhibitory activities of platelet aggregation induced by ADP and AA have been measured by Bron method. Preliminary pharmacological results showed that compounds 7g, 7h and 7n had potent inhibitory activity against platelet aggregation induced by AA, and the compound 7O showed significant inhibitory activity against platelet aggregation induced by ADP.

Shi G.-X.,Anhui University | Shi G.-X.,Anhui Academy of Chinese Medicine | Yan Y.-Y.,Anhui University | Yan Y.-Y.,Anhui Academy of Chinese Medicine | And 10 more authors.
Zhongguo Zhongyao Zazhi | Year: 2014

Objective: To in vest i gat the effect of andrographolide derivative Yanhuning ( YHN) on Candida albicans biofilms in rats. Method: The rat C. albicans hiofili s subcutaneous catheter model was established by intraperitoneally injecting YHN (40, 20, 10, 5, 2. 5 mg kg-1 ) , with the KI.C (80 mg kg-1 ) positive group as the control group. After 7 d, CFU counting and XlT assay were used to evaluate the effect of YHN on C. albicans biofilms in vivo. Scanning electron microscopy (SKM) was applied to ob-serve the morphological changes in rat biofilms intervened by YHN. The real-time fluorescence quantification PGR was adopted to detect expressions of C. albicans adhesion-related genes, such as ALS1 , ALS3, HWP1 , EAP1 and MP65. Result; The YHN group showed much less CM s on catheter pieces and lower XTT metabolic activity than the blank group, with dosage dependence. SKM also showed that YHN could obviously decrease C. albicans adhesion on subcutaneous catheters in rats. According to qRT-PCR s results, YHN can down-regulate expressions of ALS1 , ALS3, WW PI , EAP1 and MP65. Conclusion; YHN could inhibit C. albicans biofilms in rats.

Chen Y.-L.,Anhui University | Chen Y.-L.,China Pharmaceutical University | Gui S.-Y.,Anhui University | Gui S.-Y.,Anhui Academy of Chinese Medicine | And 3 more authors.
Yaoxue Xuebao | Year: 2016

Phytantriol (PT), ethanol (ET) and water were used to prepare in situ cubic liquid crystal (ISV2). The pseudo-ternary phase diagram of PT-ET-water was constructed and isotropic solution formulations were chosen for further optimization. The physicochemical properties of isotropic solution formulations were evaluated to optimize the composition of ISV2. In situ hexagonal liquid crystals (ISH2) were prepared based on the composition of ISV2 with the addition of Vitamin E acetate (VitEA) and the amount of VitEA was optimized by in vitro release behavior. The phase structures of liquid crystalline gels formed by ISV2 and ISH2 in excess water were confirmed by crossed polarized light microscopy and small angle X-ray scattering, respectively. Rheological properties of ISV2 and ISH2 were studied by a DHR-2 rheometer. In vitro drug release studies were conducted by using a dialysis membrane diffusion method. Pharmacokinetics was investigated by determination of sinomenine hydrochloride (SMH) concentration in synovial membrane after intra-articular injection of SMH-loaded ISH2 in adjuvant-induced arthritis rats. The optimal ISV2 (PT/ET/water, 64:16:20, w/w/w) loaded with 6 mg·g-1 of SMH showed a suitable pH, injectable and formed a cubic liquid crystalline gel in situ with minimum water absorption in the shortest time. The optimal ISV2 was able to sustain the drug release for 144 h. The optimal ISH2 system was prepared by addition of 5% VitEA into PT in the optimal ISV2 system. This ISH2 (PT/VitEA/ET/water, 60.8:3.2:16:20, w/w/w/w) was an injectable isotropic solution with suitable pH. The new ISH2 was able to sustain the drug release for more than 240 h. Local pharmacokinetics study indicated that the retention time and AUC0-∞ of ISH2 group were increased significantly compared with that of SMH solution group and the AUC0-∞ of ISH2 group was 6.01 times higher than that of SMH solution group. The developed ISH2 was suitable for intra-articular injection that may apply to patients in the treatment of rheumatoid arthritis.

Huang W.-J.,Anhui University | Huang W.-J.,Anhui Academy of Chinese Medicine | Xie D.,Anhui University | Xie D.,Anhui Academy of Chinese Medicine | And 8 more authors.
Yaoxue Xuebao | Year: 2016

Six novel ligustrazine chalcone aromatic oxygen alkyl acids compounds and two pyridine chalcone aromatic oxygen alkyl acids ester compounds were synthesized according to the traditional Chinese medicine theory removing blood stasis. The structures of target compounds were identified by IR, NMR and ESI-MS. The inhibitory activities of platelet aggregation induced by adenosine diphosphate (ADP) and arachidonic acid (AA) were measured by the liver microsomal incubation method in vitro. Hypolipidemic activities of compounds were tested in vivo for better inhibitory activities of platelet aggregation. Preliminary pharmacological results showed that compounds 7c, 8a and 11a had potent inhibitory activity against platelet aggregation induced by AA, compounds 7c, 7d, 8a and 11b showed significant inhibitory activity against platelet aggregation induced by ADP. Compounds 7c and 8a exhibited good hypolipidemic activities in high-fat-diet (HFD) induced hyperlipidemia C57/BL6 mice and worthy for further investigation.

Yu L.,Anhui University | Yu L.,Anhui Academy of Chinese Medicine | Wang N.,Anhui University | Wang N.,Anhui Academy of Chinese Medicine | And 9 more authors.
Neurochemistry International | Year: 2014

In the pathogenesis of cerebral ischemia, glutamate excitotoxicity activates N-methyl-d-aspartate (NMDA) receptors which induce calcium influx and oxidative stress. Muscone exerts potent neuroprotective activities on cerebral ischemia. However, its underlying mechanism is yet to be elucidated. In this study, we demonstrated that pretreatment with muscone in PC12 cells markedly ameliorated the loss of cell viability, mitochondrial membrane potential (MMP) collapse, the release of lactate dehydrogenase (LDH), Ca2+ overload, reactive oxygen species (ROS) generation, and cell apoptosis induced by glutamate. Furthermore, muscone also decreased NR1 (NMDA receptor subunit 1) protein expression, the ratio of Bax/Bcl-2 protein expression and prevented activitation of Ca2+/calmodulin-dependent protein kinase type II (CaMKII) and ASK1/JNK/p38 signaling pathways elicited by glutamate in PC12 cells. In conclusion, our results provided novel evidence that muscone protected PC12 cells against glutamate-induced apoptosis by attenuating ROS generation and Ca2+ influx, via NR1 and CaMKII-depended ASK-1/JNK/p38 signaling pathways. © 2014 Elsevier Ltd. All rights reserved.

Wu D.-Q.,Anhui University | Wu D.-Q.,Anhui Academy of Chinese Medicine | Huang W.-F.,Anhui University | Huang W.-F.,Anhui Academy of Chinese Medicine | And 6 more authors.
Zhongguo Zhongyao Zazhi | Year: 2015

Sodium houttuyfonate (SH) is a derivative of effective component of a Chinese material medica, Houttuynia cordata, which is applied in anti-infection of microorganism But, the antimicrobial mechanisms of SH still remain unclear. Here, we firstly discovered that SH effectively inhibits the three types of virulence related motility of Pseudomonas aeruginosa, i.e., swimming, twitching and swarming. The plate assay results showed that the inhibitory action of SH against swimming and twitching in 24 h and swarming in 48 h is dose-dependent; and bacteria nearly lost all of the motile activities under the concentration of 1 × minimum inhibitory concentration (MIC) (512 mg · L-1) same as azithromycin positive group (1 × MIC, 16 mg · L-1). Furthermore, we found that the expression of structural gene flgB and pilG is down-regulated by SH, which implies that inhibitory mechanism of SH against motility of P. aeruginosa may be due to the inhibition of flagella and pili bioformation of P. aeruginosa by SH. Therefore, our presented results firstly demonstrate that SH effectively inhibits the motility activities of P. aeruginosa, and suggest that SH could be a promising antipseudomonas agents in clinic.

Yu N.-J.,Anhui University | Yu N.-J.,Anhui Academy of Chinese Medicine | Peng D.-Y.,Anhui University | Peng D.-Y.,Anhui Academy of Chinese Medicine | And 8 more authors.
Zhongguo Zhongyao Zazhi | Year: 2015

Objective: To optimize the method in the Chinese Pharmacopoeia for determining Salviae Miltiorrhizae Radix et Rhizoma. Method: Tanshinone IIA and salvianolic acid B were selected as the index in optimization of the sample preparation method of Salviae Miltiorrhizae Radix et Rhizoma in Chinese Pharmacopoeia. Orthogonal test was used to optimize the extraction process of Salviae Miltiorrhizae Radix et Rhizoma, and concentration of contents were detected by high performance liquid chromatography method. A detection of using methanol-water(85: 15) at wavelength of 270 nm was employed for tanshinone IIA, and a detection of using methanolacetonitrile-formic acid-water (30: 10: 1: 59) at wavelength of 286 nm was employed for salvianolic acid B. Result: The optimized extraction process of tanshinone IIA and salvianolic acid B was: extracted by 90% methanol and reflux twice(0.5 h each time) at 75 °C, extracted by 70% methanol and reflux twice(1.5 h each time) at 75 °C, respectively. Conclusion: Optimized extraction and determination methods could be used to reflect the content of tanshinone IIA and salvianolic acid B in Salviae Miltiorrhizae Radix et Rhizoma more accurately and efficiently.

Guo J.,Anhui University | Jiang X.,Anhui University | Gui S.,Anhui University | Gui S.,Anhui Academy of Chinese Medicine
International Journal of Nanomedicine | Year: 2016

Inflammatory bowel disease (IBD), which includes ulcerative colitis and Crohn’s disease, is a chronic, recrudescent disease that invades the gastrointestinal tract, and it requires surgery or lifelong medicinal therapy. The conventional medicinal therapies for IBD, such as anti-inflammatories, glucocorticoids, and immunosuppressants, are limited because of their systemic adverse effects and toxicity during long-term treatment. RNA interference (RNAi) precisely regulates susceptibility genes to decrease the expression of proinflammatory cytokines related to IBD, which effectively alleviates IBD progression and promotes intestinal mucosa recovery. RNAi molecules generally include short interfering RNA (siRNA) and microRNA (miRNA). However, naked RNA tends to degrade in vivo as a consequence of endogenous ribonucleases and pH variations. Furthermore, RNAi treatment may cause unintended off-target effects and immunostimulation. Therefore, nanovectors of siRNA and miRNA were introduced to circumvent these obstacles. Herein, we introduce non-viral nanosystems of RNAi molecules and discuss these systems in detail. Additionally, the delivery barriers and challenges associated with RNAi molecules will be discussed from the perspectives of developing efficient delivery systems and potential clinical use. © 2016 Guo et al.

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