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O41 Traditional Chinese medicine Liuwei Dihuang decoction and its new formula, LW-AFC, as promising therapies against Alzheimer’s disease: Pharmacological actions and mechanisms
Abstracts / Biochemical Pharmacology 139 (2017) 105–141
arterioles, and reduced perivascular macrophage and lymphocyte infiltration in lungs. Conclusion: The present findings indicate that the EPA:DHA 6:1 has a cardioprotective effect in PAH by preventing right ventricular failure, pulmonary artery and arterioles remodelling and endothelial dysfunction, most likely by preventing vascular oxidative stress. doi:10.1016/j.bcp.2017.06.105
Session 14: Addressing the convergence between nutraceuticals, TCM and western medicine O41 Traditional Chinese medicine Liuwei Dihuang decoction and its new formula, LW-AFC, as promising therapies against Alzheimer’s disease: Pharmacological actions and mechanisms Wenxia Zhou, Jianhui Wang, Xiaorui Cheng, Yan Huang Beijing Institute of Pharmacology and Toxicology, Beijing, China Alzheimer’s disease (AD) is the leading cause of dementia that affects millions of elderly people worldwide. The currently available therapies have limited efficacy. Liuwei Dihuang decoction (LW), a classical traditional Chinese medicine (TCM) formula, has long been used to treat various diseases, including dementia. A large number of pharmacological studies have shown that LW has beneficial effects on AD. LW-AFC is a new formula consisting of the main active components prepared from LW. In this study, we found that administration of LW-AFC significantly improved behavioural performances in spontaneous locomotor activity, object recognition memory, spatial learning and memory, passive and active avoidance impairment in some AD mouse models, such as SAMP8 and APP/PS1 transgenic mice. Meanwhile, the impairments of long-term potentiation (LTP) were significantly ameliorated by LW-AFC administration in SAMP8 mice, as well as in the corticosterone-induced LTP inhibition mouse model. These effects were companied by an alleviation in neuron loss in the hippocampus, suppression of A-beta deposition in the brain, and a reduction in the concentration of Abeta42 in the hippocampus and plasma of APP/PS1 mice. The restoring and correcting of imbalance in the hypothalamic-pituitary-adrenal (HPA) and hypothalamic-pituitary-gonadal (HPG) axes, the disorder of lymphocyte subsets, and the abnormalities in cytokine production were also observed in LW-AFC treated SAMP8 and APP/PS1 mice. These findings indicated that LW-AFC ameliorated the behavioural and pathological deterioration of AD mice via the restoration of the NIM network in a holistic way, suggesting that LW-AFC might be a promising therapy for AD. doi:10.1016/j.bcp.2017.06.106
O42 MAM, a natural naphthoquinone, induces necroptosis in cancer cells Xiuping Chen, Wen Sun University of Macau, Macau, China Necroptosis is a recently identified programmed cell death. Here, we reported that MAM, a natural naphthoquinone isolated from Huzhang, caused hydrogen peroxide dependent activation of JNK and induced the expression of iNOS, thereby leading to nitric oxide generation in A549 lung cancer cells. In addition, a decrease in GSH/GSSG levels accompanied by increased ROS production was also observed. Reversal of ROS generation and cell death in GSH pre-
123
treated cells indicated the involvement of GSH depletion in MAM mediated cytotoxicity. Furthermore, MAM treated HCT116 and HT29 cells, were characterised by mitochondrial ROS elevation, mitochondrial depolarization and ATP depletion. This process was identified as necroptosis, based on the experiments showing that MAM-induced cell death was attenuated by RIP1 inhibitor necrostatin-1s, siRNA-mediated gene silencing of RIP1 and RIP3. RIP1/RIP3 complex triggered necroptosis via the accumulation of cytosolic calcium, followed by sustained JNK activation. Finally, we determined that both the calcium chelator BAPTA-AM and JNK inhibitor SP600125 could ameliorate mitochondrial ROS elevation, mitochondrial depolarization and ATP depletion. In addition, MAM induced necroptosis was independent of TNFa, p53, MLKL, and LMP. By demonstrating that RIP1/RIP3 complex triggered cytosolic calcium accumulation is a critical mediator in MAM-induced necroptosis by sustained activation of JNK, our study provides new insights into the molecular regulation of necroptosis in human colon cancer cells. In summary, MAM induced necroptosis in cancer cells. Acknowledgments: This study was supported by the Science and Technology Development Fund of Macau (FDCT) (078/2016/A2) and the Research Fund of University of Macau (MYRG2016-00043ICMS-QRCM). doi:10.1016/j.bcp.2017.06.107
O43 The effects and mechanisms of two main constituents from Glycyrrhiza uralensis Fisch-induced autophagy in non-small cell lung cancer Zheng-Hai Tang, Le-Le Zhang, Xin Chen, Xiuping Chen, Yitao Wang, Jin-Jian Lu State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macau, China The Glycyrrhiza uralensis Fisch, one of the famous Chinese medicinal herbs, has been widely used in anti-cancer prescription. We found that glycerrhetinic acid (GA, triterpenoid) and licochalcone A (LCA, flavonoid), two primary constituents of Glycyrrhiza uralensis Fisch, induced cell proliferative inhibition, apoptosis, and autophagy in non-small cell lung cancer (NSCLC) A549 and NCI-H1299 cells. Combined treatment with chloroquine further enhanced GA- or LCA-induced expression of LC3-II and more red-fluorescent puncta than green ones were observed in GA- or LCA-treated cells with transfection of mRFP-EGFP-LC3, suggesting that GA and LCA induces autophagic flux in NSCLC cells. Inhibition of autophagy enhanced GA-induced cell proliferative inhibition and apoptosis in NSCLC cells. In contrast, LCA-induced cell proliferative inhibition and apoptosis were not significantly altered after autophagy inhibition in NSCLC cells. The JNK and IRE1a were activated after incubation with GA. Pretreatment with the JNK inhibitor SP600125 or silencing of the JNK pathway by siRNA of JNK or c-jun decreased GA-induced autophagy. Moreover, the GA-induced JNK pathway activation and autophagy were decreased by IRE1a knockdown, suggesting that GA induces autophagy through activation of IRE1a-JNK/c-jun pathway. LCA increased the expression of CHOP, and knockdown of CHOP reversed LCA-induced autophagy, suggesting that LCA-induced autophagy is due to induction of CHOP expression. Collectively, both GA and LCA induced cell proliferative inhibition, apoptosis, and autophagy in NSCLC, and inhibition of autophagy might be an effective strategy to enhance the anti-NSCLC effects of Glycyrrhiza uralensis Fisch contained prescriptions. doi:10.1016/j.bcp.2017.06.108
arterioles, and reduced perivascular macrophage and lymphocyte infiltration in lungs. Conclusion: The present findings indicate that the EPA:DHA 6:1 has a cardioprotective effect in PAH by preventing right ventricular failure, pulmonary artery and arterioles remodelling and endothelial dysfunction, most likely by preventing vascular oxidative stress. doi:10.1016/j.bcp.2017.06.105
Session 14: Addressing the convergence between nutraceuticals, TCM and western medicine O41 Traditional Chinese medicine Liuwei Dihuang decoction and its new formula, LW-AFC, as promising therapies against Alzheimer’s disease: Pharmacological actions and mechanisms Wenxia Zhou, Jianhui Wang, Xiaorui Cheng, Yan Huang Beijing Institute of Pharmacology and Toxicology, Beijing, China Alzheimer’s disease (AD) is the leading cause of dementia that affects millions of elderly people worldwide. The currently available therapies have limited efficacy. Liuwei Dihuang decoction (LW), a classical traditional Chinese medicine (TCM) formula, has long been used to treat various diseases, including dementia. A large number of pharmacological studies have shown that LW has beneficial effects on AD. LW-AFC is a new formula consisting of the main active components prepared from LW. In this study, we found that administration of LW-AFC significantly improved behavioural performances in spontaneous locomotor activity, object recognition memory, spatial learning and memory, passive and active avoidance impairment in some AD mouse models, such as SAMP8 and APP/PS1 transgenic mice. Meanwhile, the impairments of long-term potentiation (LTP) were significantly ameliorated by LW-AFC administration in SAMP8 mice, as well as in the corticosterone-induced LTP inhibition mouse model. These effects were companied by an alleviation in neuron loss in the hippocampus, suppression of A-beta deposition in the brain, and a reduction in the concentration of Abeta42 in the hippocampus and plasma of APP/PS1 mice. The restoring and correcting of imbalance in the hypothalamic-pituitary-adrenal (HPA) and hypothalamic-pituitary-gonadal (HPG) axes, the disorder of lymphocyte subsets, and the abnormalities in cytokine production were also observed in LW-AFC treated SAMP8 and APP/PS1 mice. These findings indicated that LW-AFC ameliorated the behavioural and pathological deterioration of AD mice via the restoration of the NIM network in a holistic way, suggesting that LW-AFC might be a promising therapy for AD. doi:10.1016/j.bcp.2017.06.106
O42 MAM, a natural naphthoquinone, induces necroptosis in cancer cells Xiuping Chen, Wen Sun University of Macau, Macau, China Necroptosis is a recently identified programmed cell death. Here, we reported that MAM, a natural naphthoquinone isolated from Huzhang, caused hydrogen peroxide dependent activation of JNK and induced the expression of iNOS, thereby leading to nitric oxide generation in A549 lung cancer cells. In addition, a decrease in GSH/GSSG levels accompanied by increased ROS production was also observed. Reversal of ROS generation and cell death in GSH pre-
123
treated cells indicated the involvement of GSH depletion in MAM mediated cytotoxicity. Furthermore, MAM treated HCT116 and HT29 cells, were characterised by mitochondrial ROS elevation, mitochondrial depolarization and ATP depletion. This process was identified as necroptosis, based on the experiments showing that MAM-induced cell death was attenuated by RIP1 inhibitor necrostatin-1s, siRNA-mediated gene silencing of RIP1 and RIP3. RIP1/RIP3 complex triggered necroptosis via the accumulation of cytosolic calcium, followed by sustained JNK activation. Finally, we determined that both the calcium chelator BAPTA-AM and JNK inhibitor SP600125 could ameliorate mitochondrial ROS elevation, mitochondrial depolarization and ATP depletion. In addition, MAM induced necroptosis was independent of TNFa, p53, MLKL, and LMP. By demonstrating that RIP1/RIP3 complex triggered cytosolic calcium accumulation is a critical mediator in MAM-induced necroptosis by sustained activation of JNK, our study provides new insights into the molecular regulation of necroptosis in human colon cancer cells. In summary, MAM induced necroptosis in cancer cells. Acknowledgments: This study was supported by the Science and Technology Development Fund of Macau (FDCT) (078/2016/A2) and the Research Fund of University of Macau (MYRG2016-00043ICMS-QRCM). doi:10.1016/j.bcp.2017.06.107
O43 The effects and mechanisms of two main constituents from Glycyrrhiza uralensis Fisch-induced autophagy in non-small cell lung cancer Zheng-Hai Tang, Le-Le Zhang, Xin Chen, Xiuping Chen, Yitao Wang, Jin-Jian Lu State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macau, China The Glycyrrhiza uralensis Fisch, one of the famous Chinese medicinal herbs, has been widely used in anti-cancer prescription. We found that glycerrhetinic acid (GA, triterpenoid) and licochalcone A (LCA, flavonoid), two primary constituents of Glycyrrhiza uralensis Fisch, induced cell proliferative inhibition, apoptosis, and autophagy in non-small cell lung cancer (NSCLC) A549 and NCI-H1299 cells. Combined treatment with chloroquine further enhanced GA- or LCA-induced expression of LC3-II and more red-fluorescent puncta than green ones were observed in GA- or LCA-treated cells with transfection of mRFP-EGFP-LC3, suggesting that GA and LCA induces autophagic flux in NSCLC cells. Inhibition of autophagy enhanced GA-induced cell proliferative inhibition and apoptosis in NSCLC cells. In contrast, LCA-induced cell proliferative inhibition and apoptosis were not significantly altered after autophagy inhibition in NSCLC cells. The JNK and IRE1a were activated after incubation with GA. Pretreatment with the JNK inhibitor SP600125 or silencing of the JNK pathway by siRNA of JNK or c-jun decreased GA-induced autophagy. Moreover, the GA-induced JNK pathway activation and autophagy were decreased by IRE1a knockdown, suggesting that GA induces autophagy through activation of IRE1a-JNK/c-jun pathway. LCA increased the expression of CHOP, and knockdown of CHOP reversed LCA-induced autophagy, suggesting that LCA-induced autophagy is due to induction of CHOP expression. Collectively, both GA and LCA induced cell proliferative inhibition, apoptosis, and autophagy in NSCLC, and inhibition of autophagy might be an effective strategy to enhance the anti-NSCLC effects of Glycyrrhiza uralensis Fisch contained prescriptions. doi:10.1016/j.bcp.2017.06.108