Background Baicalin has a significant anti-inflammation effect and is widely used in the clinical treatment of stroke. Most of the studies of Toll-like receptor 2/4 （TLR2/4） during cerebral ischemia had defined the...Background Baicalin has a significant anti-inflammation effect and is widely used in the clinical treatment of stroke. Most of the studies of Toll-like receptor 2/4 （TLR2/4） during cerebral ischemia had defined their specific expressions in microglia in hippocampus tissue. To explore the targets of baicalin in stroke, we detected the expressions of TLR2/4 in vitro/vivo.Methods By constructing a cerebral ischemia-reperfusion model in vivo and glucose oxygen deprivation model, we successfully induced neuron damage, then added baicalin and detected expressions of TLR2/4, nuclear factor-kB （NF-kB）, tumor necrosis factor-alpha （TNFα）, and interleukin-1β （IL-1β） in mRNA level and protein level.Results We found distinct upregulations of TLR2/4 and TNFα in both mRNA level and protein level in PC12 cells and primary neurons. Moreover, TLR2/4 and TNFα expressions were significantly higher in mice hippocampus treated with cerebral ischemia-reperfusion. Baicalin could downregulate the expressions of TLR2/4 and TNFα in the damaged cells and mice hippocampus effectively.Conclusions Neurons could respond to the damage and activate the related signal pathway directly. TLR2/4 responsed to the damage and sent the signal to downstream factor TNFα through activating NF-kB. Baicalin could inhibit the inflammatory reaction in neuron damage and TLR might be its targets, which explained why baicalin could widely be used in the clinical treatment of stroke.展开更多
Angelica dahurica(A. dahurica) is a traditional Chinese medicinal plant being used in clinical practice. The present study demonstrated that A. dahurica could reduce white-fat weight in high-fat-diet hyperlipidemic mi...Angelica dahurica(A. dahurica) is a traditional Chinese medicinal plant being used in clinical practice. The present study demonstrated that A. dahurica could reduce white-fat weight in high-fat-diet hyperlipidemic mice, decrease total cholesterol and triglyceride concentrations in the livers of both high-fat-diet and Triton WR1339 induced hyperlipidemic mice, and enhance the total hepatic lipase activities of them. These findings were further supported by the results derived from the experiments with Hep G2 cells in vitro. In addition, the proteins related to lipids metabolism were investigated using LC-MS/MS, indicating that genes of lipid metabolism and lipid transport were regulated by A. dhurica. The results from LC-MS/MS were further conformed by Western blot and real time PCR assays. A. dahurica could down-regulate the expression of catalase(CAT) and sterol carrier protein2(SCP2) and up-regulate the expression of lipid metabolism related genes-lipase member C(LIPC) and peroxisome proliferator-activated receptor gamma(PPARγ). In the Triton WR1339 mouse liver and Hep G2 cells in vitro, A. dahurica was able to increase the expression of LIPC and PPARγ, confirming the results from in vivo experiments. Imperatorin showed the same activity as A. dahurica, suggesting it was one of the major active ingredients of the herb. In conclusion, our work represented a first investigation demonstrating that A. dahurica was able to regulate lipid metabolism and could be developed as a novel approach to fighting against fatty liver and obesity.展开更多
Heat stress can stimulate an increase in body temperature, which is correlated with increased expression of heat shock protein 70 （HSP70） and tumor necrosis factor a （TNFa）. The exact mechanism underlying the HSP7...Heat stress can stimulate an increase in body temperature, which is correlated with increased expression of heat shock protein 70 （HSP70） and tumor necrosis factor a （TNFa）. The exact mechanism underlying the HSP70 and TNFa induction is unclear. Berberine （BBR） can significantly inhibit the temperature rise caused by heat stress, but the mechanism responsible for the BBR effect on HSP70 and TNFa signaling has not been investigated. The aim of the present study was to explore the relationship between the expression of HSP70 and TNFa and the effects of BBR under heat conditions, using in vivo and in vitro models. The expression levels of HSP70 and YNFa were determined using RT-PCR and Western blotting analyses. The results showed that the levels of HSP70 and TNFa were ap-regulated under heat conditions （40 ~C）. HSP70 acted as a chaperone to maintain TNFa homeostasis with rising the temperature, but knockdown of HSP70 could not down-regulate the level of TNFa. Furthermore, TNFa could not influence the expression of HSP70 under aormal and heat conditions. BBR targeted both HSP70 and TNFa by suppressing their gene transcription, thereby decreasing body temperature under heat conditions. In conclusion, BBR has a potential to be developed as a therapeutic strategy for suppressing the thermal effects in hot environments.展开更多
Brazilein is reported to have immunosuppressive effect on cardiovascular and cerebral-vascular diseases. The essential roles of innate immunity in cerebral ischemia are increasingly identified, but no studies concerni...Brazilein is reported to have immunosuppressive effect on cardiovascular and cerebral-vascular diseases. The essential roles of innate immunity in cerebral ischemia are increasingly identified, but no studies concerning the influence of brazilein on the innate immunity receptors have been reported. The present study was designed to investigate the regulation of NOD2 （Nucleotide-binding oligomerization domain-containing protein 2） by brazilein for its protection of neuron in cerebral ischemia in vivo and oxygen-glucose deprivation in vitro. The results showed that brazilein could reverse the elevated expression of NOD2 and TNFa （tumor necrosis factor alpha） elicited by cerebral ischemia and reperfusion. This reduction could also be detected in normal mice and C 17.2 cells, indicating that this suppressive effect of brazilein was correlated with NOD2. The results from GFP reporter plasmid assay suggested brazilein inhibited NOD2 gene transcription. In conclusion, brazilein could attenuate NOD2 and TNFα expression in cerebral ischemia and NOD2 may be one possible target of brazilein for its immune suppressive effect in neuro-inflammation.展开更多
Brazilein is an active small molecular compound extracted from Caesalpinia sappan L. with favorable pharmacological properties on immune system, cardiovascular system, and nervous system. C. sappan has been used as a ...Brazilein is an active small molecular compound extracted from Caesalpinia sappan L. with favorable pharmacological properties on immune system, cardiovascular system, and nervous system. C. sappan has been used as a traditional medicine in China for hundreds of years for various diseases. However, the general reproductive toxicity of brazilein is still unknown. The purpose of the present study was to thoroughly evaluate the general reproductive toxicity of brazilein in ICR mice to support the future drug development and modernization of this potent traditional Chinese medicine. The results showed that, although no apparent toxicity on the reproducibility of the male was observed, brazilein might cause considerable risks to the fetuses and females as indicated by the ratios of dead fetuses and reabsorptions. In conclusion, our results from the present study provided some useful insights about the safety profile of brazilein, suggesting that brazilein should be used with caution in pregnant women.展开更多
基金This work was supported by grants from the National Natural Science Foundation of China （No. 30973896, No. 30801523, and No. 81073092） and the National S＆T Major Special Project for New Drug R＆D Program of China （No. 2009ZX09103-301, No. 2009ZX09502, No. 2009ZX09308-003, and No. 2011ZX09101- 002-11）.Acknowledgments： We thank specially all our colleagues in our laboratory.
文摘Background Baicalin has a significant anti-inflammation effect and is widely used in the clinical treatment of stroke. Most of the studies of Toll-like receptor 2/4 （TLR2/4） during cerebral ischemia had defined their specific expressions in microglia in hippocampus tissue. To explore the targets of baicalin in stroke, we detected the expressions of TLR2/4 in vitro/vivo.Methods By constructing a cerebral ischemia-reperfusion model in vivo and glucose oxygen deprivation model, we successfully induced neuron damage, then added baicalin and detected expressions of TLR2/4, nuclear factor-kB （NF-kB）, tumor necrosis factor-alpha （TNFα）, and interleukin-1β （IL-1β） in mRNA level and protein level.Results We found distinct upregulations of TLR2/4 and TNFα in both mRNA level and protein level in PC12 cells and primary neurons. Moreover, TLR2/4 and TNFα expressions were significantly higher in mice hippocampus treated with cerebral ischemia-reperfusion. Baicalin could downregulate the expressions of TLR2/4 and TNFα in the damaged cells and mice hippocampus effectively.Conclusions Neurons could respond to the damage and activate the related signal pathway directly. TLR2/4 responsed to the damage and sent the signal to downstream factor TNFα through activating NF-kB. Baicalin could inhibit the inflammatory reaction in neuron damage and TLR might be its targets, which explained why baicalin could widely be used in the clinical treatment of stroke.
基金supported in part by National Natural Science Foundation of China(Nos.81473321 and 81374006)the National Key Technology R&D Program of China(No.2012BAI29B02)the National S&T Major Special Project for New Drug R&D Program of China(Nos.2012ZX09102-201-008,2012ZX09103-201-041,and 2011ZX09101-002-11)
文摘Angelica dahurica(A. dahurica) is a traditional Chinese medicinal plant being used in clinical practice. The present study demonstrated that A. dahurica could reduce white-fat weight in high-fat-diet hyperlipidemic mice, decrease total cholesterol and triglyceride concentrations in the livers of both high-fat-diet and Triton WR1339 induced hyperlipidemic mice, and enhance the total hepatic lipase activities of them. These findings were further supported by the results derived from the experiments with Hep G2 cells in vitro. In addition, the proteins related to lipids metabolism were investigated using LC-MS/MS, indicating that genes of lipid metabolism and lipid transport were regulated by A. dhurica. The results from LC-MS/MS were further conformed by Western blot and real time PCR assays. A. dahurica could down-regulate the expression of catalase(CAT) and sterol carrier protein2(SCP2) and up-regulate the expression of lipid metabolism related genes-lipase member C(LIPC) and peroxisome proliferator-activated receptor gamma(PPARγ). In the Triton WR1339 mouse liver and Hep G2 cells in vitro, A. dahurica was able to increase the expression of LIPC and PPARγ, confirming the results from in vivo experiments. Imperatorin showed the same activity as A. dahurica, suggesting it was one of the major active ingredients of the herb. In conclusion, our work represented a first investigation demonstrating that A. dahurica was able to regulate lipid metabolism and could be developed as a novel approach to fighting against fatty liver and obesity.
基金supported by the National Natural Science Foundation of China(Nos.81374006,90713043 and 81073092)
文摘Heat stress can stimulate an increase in body temperature, which is correlated with increased expression of heat shock protein 70 （HSP70） and tumor necrosis factor a （TNFa）. The exact mechanism underlying the HSP70 and TNFa induction is unclear. Berberine （BBR） can significantly inhibit the temperature rise caused by heat stress, but the mechanism responsible for the BBR effect on HSP70 and TNFa signaling has not been investigated. The aim of the present study was to explore the relationship between the expression of HSP70 and TNFa and the effects of BBR under heat conditions, using in vivo and in vitro models. The expression levels of HSP70 and YNFa were determined using RT-PCR and Western blotting analyses. The results showed that the levels of HSP70 and TNFa were ap-regulated under heat conditions （40 ~C）. HSP70 acted as a chaperone to maintain TNFa homeostasis with rising the temperature, but knockdown of HSP70 could not down-regulate the level of TNFa. Furthermore, TNFa could not influence the expression of HSP70 under aormal and heat conditions. BBR targeted both HSP70 and TNFa by suppressing their gene transcription, thereby decreasing body temperature under heat conditions. In conclusion, BBR has a potential to be developed as a therapeutic strategy for suppressing the thermal effects in hot environments.
基金supported by the National S&T Major Special Project for New Drug R&D Program of China(Nos.2012ZX09102-201-008,2012ZX09103-201-041,and 2011ZX09101-002-11)National Natural Science Foundation of China(Nos.81374006 and 81073092)
文摘Brazilein is reported to have immunosuppressive effect on cardiovascular and cerebral-vascular diseases. The essential roles of innate immunity in cerebral ischemia are increasingly identified, but no studies concerning the influence of brazilein on the innate immunity receptors have been reported. The present study was designed to investigate the regulation of NOD2 （Nucleotide-binding oligomerization domain-containing protein 2） by brazilein for its protection of neuron in cerebral ischemia in vivo and oxygen-glucose deprivation in vitro. The results showed that brazilein could reverse the elevated expression of NOD2 and TNFa （tumor necrosis factor alpha） elicited by cerebral ischemia and reperfusion. This reduction could also be detected in normal mice and C 17.2 cells, indicating that this suppressive effect of brazilein was correlated with NOD2. The results from GFP reporter plasmid assay suggested brazilein inhibited NOD2 gene transcription. In conclusion, brazilein could attenuate NOD2 and TNFα expression in cerebral ischemia and NOD2 may be one possible target of brazilein for its immune suppressive effect in neuro-inflammation.
基金supported by the National Science&Technology Major Special Project for New Drug R&D Program of China(No.2012ZX09102-201-008)
文摘Brazilein is an active small molecular compound extracted from Caesalpinia sappan L. with favorable pharmacological properties on immune system, cardiovascular system, and nervous system. C. sappan has been used as a traditional medicine in China for hundreds of years for various diseases. However, the general reproductive toxicity of brazilein is still unknown. The purpose of the present study was to thoroughly evaluate the general reproductive toxicity of brazilein in ICR mice to support the future drug development and modernization of this potent traditional Chinese medicine. The results showed that, although no apparent toxicity on the reproducibility of the male was observed, brazilein might cause considerable risks to the fetuses and females as indicated by the ratios of dead fetuses and reabsorptions. In conclusion, our results from the present study provided some useful insights about the safety profile of brazilein, suggesting that brazilein should be used with caution in pregnant women.
