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Ginsenoside Rb1 improves brain,lung,and intestinal barrier damage in middle cerebral artery occlusion/reperfusion(MCAO/R)micevia the PPARγsignaling pathway

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Chinese Journal of Natural Medicines 2022年第8期20卷 561-571页

作者： SU Lin-Jie REN Yu-Chuan CHEN Zhuo MA Hui-Fen ZHENG Fan LI Fang ZHANG Yuan-Yuan GONG Shuai-Shuai kou jun-ping Jiangsu Key Laboratory of TCM Evaluation and Translational Research Department of Pharmacology of Chinese Materia MedicaSchool of Traditional Chinese PharmacyChina Pharmaceutical UniversityNanjing 211198China

Ischemic stroke causes brain inflammation and multi-organ injury,which is closely associated with the peroxisome proliferator-activated receptor-gamma(PPARγ)signaling pathway.Recent studies have indicated that ginsenoside Rb1(GRb1)can protect the integrity of the blood-brain barrier after stroke.In the current study,a mouse model of middle cerebral artery occlusion/reperfusion(MCAO/R)was established to determine whether GRb1 can ameliorate brain/lung/intestinal barrier damage via the PPARγsignaling pathway.Staining(2,3,5-triphenyltetrazolium chloride,hematoxylin,and eosin)and Doppler ultrasonography were employed to detect pathological changes.Endothelial breakdown was investigated with the leakage of Evans Blue dye and the expression of TJs(tight junctions)and AJs(adherent junctions).Western blot and immunofluorescence were used to determine the levels of cell junction proteins,PPARγand NF-κB.Results showed that GRb1 significantly mitigated multi-organ injury and increased the expression of cerebral microvascular,pulmonary vascular,and intestinal epithelial connexins.In brain,lung,and intestinal tissues,GRb1 activated PPARγ,decreased the levels of phospho-NF-κB p65,and inhibited the production of proinflammatory cytokines,thereby maintaining barrier permeability.However,co-treatment with GRb1 and the PPARγantagonist GW9662 reversed the barrier-protective effect of GRb1.These findings indicated that GRb1 can improve stroke-induced brain/lung/intestinal barrier damagevia the PPARγpathway.

关键词： Focal ischemic stroke Brain/lung/intestinal barriers Ginsenoside Rb1 PPARγ

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An optimized micro-assay of myosin Ⅱ ATPase activity based on the molybdenum blue method and its application in screening natural product inhibitors

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Chinese Journal of Natural Medicines 2016年第6期14卷 421-426页

作者： CHEN Hong-Lin ZHAO Jing ZHANG Guan-jun kou jun-ping YU Bo-Yang Jiangsu Key Laboratory of TCM Evaluation and Translational Research Department of Complex Prescription of TCM China Pharmaceutical University

Myosin Ⅱ plays multiple roles in physiological and pathological functions through its ATPase activity. The present study was designed to optimize a micro-assay of myosin Ⅱ ATPase activity based on molybdenum blue method, using a known myosin Ⅱ ATPase inhibitor, blebbistatin. Several parameters were observed in the enzymatic reaction procedure, including the concentrations of the substrate(ATP) and calcium chloride, p H, and the reaction and incubation times. The proportion of coloration agent was also investigated. The sensitivity of this assay was compared with the malachite green method and bioluminescence method. Additionally, 20 natural compounds were studied for myosin Ⅱ ATPase inhibitory activity using the optimized method. Our results showed that ATP at the concentration of 5 mmol·L^(-1) and ammonium molybdate : stannous chloride at the ratio of 15 : 1 could greatly improve the sensitivity of this method. The IC50 of blebbistatin obtained by this method was consistent with literature. Compound 8 was screened with inhibitory activity on myosin Ⅱ ATPase. The optimized method showed similar accuracy, lower detecting limit, and wider linear range, which could be a promising approach to screening myosin Ⅱ ATPase inhibitors in vitro.

关键词： Myosin Ⅱ ATPase activity Blebbistatin Bioluminescence Natural products Malachite green

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Sheng-Mai-San attenuates contractile dysfunction and structural damage induced by chronic intermittent hypoxia in mice

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Chinese Journal of Natural Medicines 2015年第10期13卷 743-750页

作者： MO Wei-Lan CHAI Cheng-Zhi kou jun-ping YAN Yong-Qing YU Bo-Yang Department of Complex Prescription of TCM China Pharmaceutical University Jiangsu Provincial Key Laboratory for TCM Evaluation and Translational Research China Pharmaceutical University

Sheng-Mai-San(SMS), a well-known Chinese medicinal plant formula, is widely used for the treatment of cardiac diseases characterized by deficiency of Qi and Yin syndrome. A mouse chronic intermittent hypoxia(CIH) model was established to mimic the primary clinical features of deficiency of Qi and Yin syndrome. Mice experienced CIH for 28 days(nadir 7% to peak 8% oxygen, 20 min per day), resulting in left ventricle(LV) dysfunction and structure abnormalities. After administration of SMS(0.55, 1.1, and 5.5 g·kg-1·d-1) for four weeks, improved cardiac function was observed, as indicated by the increase in the ejection fraction from the LV on echocardiography. SMS also preserved the structural integrity of the LV against eccentric hypotrophy, tissue vacuolization, and mitochondrial injury as measured by histology, electron microscopy, and ultrasound assessments. Mechanistically, the antioxidant effects of SMS were demonstrated; SMS was able to suppress mitochondrial apoptosis as indicated by the reduction of several pro-apoptotic factors(Bax, cytochrome c, and cleaved caspase-3) and up-regulation of the anti-apoptosis factor Bcl-2. In conclusion, these results demonstrate that SMS treatment can protect the structure and function of the LV and that the protective effects of this formula are associated with the regulation of the mitochondrial apoptosis pathway.

关键词： Sheng-Mai-San Chronic intermittent hypoxia Contractile dysfunction Left ventricle Mitochondrial apoptosis

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Network pharmacology-based prediction and verification of the molecular targets and pathways for schisandrin against cerebrovascular disease

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Chinese Journal of Natural Medicines 2014年第4期12卷 251-258页

作者： LV Yan-Ni LI Shao-Xia ZHAI Ke-Feng kou jun-ping YU Bo-Yang State Key Laboratory of Natural Medicines China Pharmaceutical University Department of Complex Prescription of TCM China Pharmaceutical University

AIM: To illuminate the molecular targets for schisandrin against cerebrovascular disease based on the combined methods of network pharmacology prediction and experimental verification. METHOD: A protein database was established through constructing the drug-protein network from literature mining data. The protein-protein network was built through an in-depth exploration of the relationships between the proteins. The computational platform was implemented to predict and extract the sensitive sub-network with significant P-values from the protein-protein network. Then the key targets and pathways were identified from the sensitive sub-network. The most related targets and pathways were also confirmed in hydrogen peroxide(H2O2)-induced PC12 cells by Western blotting. RESULTS: Twelve differentially expressed proteins(gene names: NFKB1, RELA, TNFSF10, MAPK1, CHUK, CASP8, PIGS2, MAPK14, CREB1, IFNG, APP, and BCL2) were confirmed as the central nodes of the interaction network(45 nodes, 93 edges). The NF-κB signaling pathway was suggested as the most related pathway of schisandrin for cerebrovascular disease. Furthermore, schisandrin was found to suppress the expression and phosphorylation of IKKα, as well as p50 and p65 induced by H2O2 in PC12 cells by Western blotting. CONCLUSION: The computational platform that integrates literature mining data, protein-protein interactions, sensitive sub-network, and pathway results in identification of the NF-κB signaling pathway as the key targets and pathways for schisandrin.

关键词： Schisandrin Network pharmacology Cerebrovascular disease Molecular target NF-κB signaling pathway

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Recent advances of traditional Chinese medicine on the prevention and treatment of COVID-19

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Chinese Journal of Natural Medicines 2020年第12期18卷 881-889页

作者： DAI Yu-Jie WAN Shi-Yao GONG Shuai-Shuai LIU Jin-Cheng LI Fang kou jun-ping Jiangsu Key Laboratory of TCM Evaluation and Translational Research School of Traditional Chinese PharmacyDepartment of Pharmacology of Chinese Materia MedicaChina Pharmaceutical UniversityNanjing 211198China

Coronavirus disease-2019(COVID-19)is a new highly infectious disease caused by a novel coronavirus.Recently,the number of new cases infected pneumonia in the world continues to increase,which has aroused great concern from the international community.At present,there are no small-molecule specific anti-viral drugs for the treatment.The high mortality rate seriously threatens human health.Traditional Chinese medicine(TCM)is a unique health resource in China.The combination of TCM and Western medicine has played a positive and important role in combating COVID-19 in China.In this review,through literature mining and analysis,it was found that TCM has the potential to prevent and treat the COVID-19.Then,the network pharmacological studies demonstrated that TCM played roles of anti-virus,anti-inflammation and immunoregulation in the management of COVID-19 via multiple components acting on multiple targets and multiple pathways.Finally,clinical researches also confirmed the beneficial effects of TCM on the treatment of patients.This review may provide meaningful and useful information on further drug development of COVID-19 and other viral infectious diseases.

关键词： Novel coronavirus pneumonia Prescription of traditional Chinese medicine Single Chinese herb Active ingredients of traditional Chinese medicine Viral infection

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Advances in the antitumor activities and mechanisms of action of steroidal saponins

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Chinese Journal of Natural Medicines 2018年第10期16卷 732-748页

作者： ZHAO Ya-Zheng ZHANG Yuan-Yuan HAN Han FAN Rui-ping HU Yang ZHONG Liang kou jun-ping YU Bo-Yang Jiangsu Key Laboratory of TCM Evaluation and Translational Research Department of Complex Prescription of Traditional Chinese MedicineChina Pharmaceutical University

The steroidal saponins are one of the saponin types that exist in an unbound state and have various pharmacological activities,such as anticancer,anti-inflammatory,antiviral,antibacterial and nerves-calming properties.Cancer is a growing health problem worldwide.Significant progress has been made to understand the antitumor effects of steroidal saponins in recent years.According to reported findings,steroidal saponins exert various antitumor activities,such as inhibiting proliferation,inducing apoptosis and autophagy,and regulating the tumor microenvironment,through multiple related signaling pathways.This article focuses on the advances in domestic and foreign studies on the antitumor activity and mechanism of actions of steroidal saponins in the last five years to provide a scientific basis and research ideas for further development and clinical application of steroidal saponins.

关键词： Steroidal saponins 肿瘤机制

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Novel cytotoxic steroidal glycosides from the roots of Liriope muscari

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Chinese Journal of Natural Medicines 2015年第6期13卷 461-466页

作者： LI Yong-Wei QI Jin ZHANG Yuan-Yuan HUANG Zhen kou jun-ping ZHOU Shui-ping ZHANG Yu YU Bo-Yang State Key Laboratory of Natural Medicines Department of Complex Prescription of TCM China Pharmaceutical University Tasly Research and Development Institute Tasly Pharmaceutical Co. Ltd.

The present study was designed to investigate the chemical constituents and bioactivities of the roots of Liriope muscari(Decne.) L.H. Bailey. The compounds were isolated through various chromatography techniques, including silica gel, Sephadex LH-20, and semi-preparative HPLC. The structures were elucidated by infrared(IR), mass spectrometric(MS), 1D- and 2D-NMR analyses in comparison with reference data. In addition, the cytotoxicity of these compounds against human breast cancer MDA-MB-435 cells was evaluated by the MTT assay. Two new steroidal glycosides, 25(R, S)-ruscogenin-1-O-[β-D-fucopyranosyl(1→2)]-[ β-D-xylopyranosyl(1→3)]-β-D-glucopyranoside(Liriopem I, 1) and 25(R, S)- ruscogenin-1-O-[β-D-fucopyranosyl(1→2)]-[β-D-xylopyranosyl(1→4)]-β-D-fucopyranoside(Liriopem II, 2) and two known compounds LM-S6(3) and DT-13(4) were isolated and identified. Liriopem I(1), liriopem II(2) and DT-13(4) showed remarkable cytotoxicity with IC50 values being(0.58 ± 0.08),(0.05 ± 0.10), and(0.15 ± 0.09) μg·m L-1, respectively. In summary, compounds 1 and 2 identified in the present study exerted cytotoxicity against breast cancer cells, providing a basis for future development of these compounds as novel anticancer agents.

关键词：细胞毒性葡糖苷甾体根部葡萄麦冬 Sephadex 乳腺癌细胞

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Ge-Gen Decoction attenuates oxytocin-induced uterine contraction and writhing response: potential application in primary dysmenorrhea therapy

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Chinese Journal of Natural Medicines 2016年第2期14卷 124-132页

作者： YANG Lu CHAI Cheng-Zhi YUE Xin-Yi YAN Yan kou jun-ping CAO Zheng-Yu YU Bo-Yang Department of Complex Prescription of TCM China Pharmaceutical University Jiangsu Provincial Key Laboratory for TCM Evaluation and Translational Research China Pharmaceutical University

The uterine tetanic contraction and uterine artery blood flow reduction are possible reasons for primary dysmenorrhea(PD). In the present study, we aimed to evaluate the uterine relaxant effect and the influence on uterine artery blood velocity of Ge-Gen Decoction(GGD), a well-known Chinese herbal formula. In female ICR mice, uterine contraction was induced by oxytocin exposure following estradiol benzoate pretreatment, and the uterine artery blood velocity was detected by Doppler ultrasound. Histopathological examination of the uterine tissue samples were performed by H&E staining. Ex vivo studies demonstrated that oxytocin, posterior pituitary, or acetylcholine induced contractions in isolated mouse uterus. GGD inhibited both spontaneous and stimulated contractions. In vivo study demonstrated that GGD significantly reduced oxytocin-induced writhing responses with a maximal inhibition of 87%. Further study demonstrated that GGD normalized oxytocin-induced abnormalities of prostaglandins F_2 alpha(PGF_(2α)) and Ca^(2+) in mice. In addition, injection of oxytocin induced a decrease in uterine artery blood flow velocity. Pretreatment with GGD reversed the oxytocin response on blood flow velocity. Histopathological examination showed pretreatment with GGD alleviated inflammation and edema in the uterus when compared with the model group. Both ex vivo and in vivo results indicated that GGD possessed a significant spasmolytic effect on uterine tetanic contraction as well as improvement on uterine artery blood velocity which may involve PGF_(2α) and Ca^(2+) signaling, suggesting that GGD may have a clinic potential in PD therapy.

关键词： Primary dysmenorrheal Spasmolytic and analgesic effect Uterine contraction Uterine artery blood flow Ge-Gen Decoction

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A combination of four effective components derived from Sheng-mai san attenuates hydrogen peroxide-induced injury in PC12 cells through inhibiting Akt and MAPK signaling pathways

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Chinese Journal of Natural Medicines 2016年第7期14卷 508-517页

作者： CAO Guo-Sheng LI Shao-Xia WANG Yan XU Ying-Qiong LV Yan-Ni kou jun-ping YU Bo-Yang Jiangsu Key Laboratory of TCM Evaluation and Translational Research Department of Complex Prescription of TCM China Pharmaceutical University

The present study was designed to investigate whether a combination of four effective components derived from Sheng-mai san(SMXZF; ginsenoside Rb1: ginsenoside Rg1: DT^(–1)3: Schizandrol A as 6 : 9 : 4 : 5) could attenuate hydrogen peroxide(H_2O_2)-induced injury in PC12 cells, focusing on the Akt and MAPK pathways. The PC12 cells were exposed to H_2O_2(400 mmol·L^(–1)) for 1 h in the presence or absence of SMXZF pre-treatment for 24 h. Cell viability was measured by MTT assay. The efflux of lactate dehydrogenase(LDH), the intracellular content of malondialdehyde(MDA), the activities of superoxide dismutase(SOD), and caspase-3 were also determined. Cell apoptosis was measured by Hoechst 33342 staining and Annexin V-FITC/PI staining method. The expression of Bcl-2, Bax, cleaved caspase-3, Akt, and MAPKs were detected by Western blotting analyses. SMXZF pretreatment significantly increased the cell viability and SOD activity and improved the cell morphological changes, while reduced the levels of LDH and MDA at the concentrations of 0.1, 1 and 10 μg·m L^(–1). SMXZF also inhibited H_2O_2-induced apoptosis in PC12 cells. Moreover, SMXZF reduced the activity of caspase-3, up-regulated the protein ratio of Bcl-2 and Bax and inhibited the expression of cleaved caspase-3, p-Akt, p-p38, p-JNK and p-ERK1/2 in H_2O_2-induced PC12 cells. Co-incubation of Akt inhibitor or p38 inhibitor partly attenuated the protection of SMXZF against H_2O_2-injured PC12 cells. In conclusion, our findings suggested that SMXZF attenuated H_2O_2-induced injury in PC12 cells by inhibiting Akt and MAPKs signaling pathways, which might shed insights on its neuroprotective mechanism.

关键词： Combination of effective TCM components Sheng-mai san SMXZF PC12 cells Hydrogen peroxide Akt MAPKs

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Advances in the pharmacological activities and mechanisms of diosgenin

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Chinese Journal of Natural Medicines 2015年第8期13卷 578-587页

作者： CHEN Yan TANG You-Mei YU Su-Lan HAN Yu-Wei kou jun-ping LIU Bao-Lin YU Bo-Yang Jiangsu Key Laboratory of TCM Evaluation and Translational Research Department of Complex Prescription of TCM China Pharmaceutical University

Diosgenin, a well-known steroid sapogenin derived from plants, has been used as a starting material for production of steroidal hormones. The present review will summarize published literature concerning pharmacological potential of diosgenin, and the underlying mechanisms of actions. Diosgenin has shown a vast range of pharmacological activities in preclinical studies. It exhibits anticancer, cardiovascular protective, anti-diabetes, neuroprotective, immunomodulatory, estrogenic, and skin protective effects, mainly by inducing apoptosis, suppressing malignant transformation, decreasing oxidative stress, preventing inflammatory events, promoting cellular differentiation/proliferation, and regulating T-cell immune response, etc. It interferes with cell death pathways and their regulators to induce apoptosis. Diosgenin antagonizes tumor metastasis by modulating epithelial-mesenchymal transition and actin cytoskeleton to change cellular motility, suppressing degradation of matrix barrier, and inhibiting angiogenesis. Additionally, diosgenin improves antioxidant status and inhibits lipid peroxidation. Its anti-inflammatory activity is through inhibiting production of pro-inflammatory cytokines, enzymes and adhesion molecules. Furthermore, diosgenin drives cellular growth/differentiation through the estrogen receptor(ER) cascade and transcriptional factor PPARγ. In summary, these mechanistic studies provide a basis for further development of this compound for pharmacotherapy of various diseases.

关键词： Diosgenin Steroid sapogenin Pharmacological activities Multiple targets

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