Matrine （MT）, the effective component of Sophora fla- vescens Air, has been shown to have anti-inflammation, immune-suppressive, anti-tumor, and anti-hepatic fibrosis activities. However, the pharmacological effects...Matrine （MT）, the effective component of Sophora fla- vescens Air, has been shown to have anti-inflammation, immune-suppressive, anti-tumor, and anti-hepatic fibrosis activities. However, the pharmacological effects of MT still need to be strengthened due to its relatively low efficacy and short half-life. In the present study, we report a more effective thio derivative of MT, MD-1, and its inhibitory effects on the activation of hepatic stellate cells （HSCs） in both cell culture and animal models. Cytological experiments showed that MD-1 can inhibit the proliferation of HSC-T6 cells with a half-maximal inhibitory concentration （ICs0） of 62 pmollL. In addition, MD-1 more strongly inhibits the migration of HSC-T6 cells compared to MT and can more effectively induce G0/G1 arrest and apoptosis. Investigating the biological mechanisms underlying anti-hepatic fibrosis in the presence of MD-I, we found that MD-I can bind the epidermal growth factor receptor （EGFR） on the surface of HSC-T6 cells, which can further inhibit the phospho- rylaUon of EGFR and its downstream protein kinase B （Akt）, resulting in decreased expression of cyclin D1 and eventual inhibition of the activation of HSC-T6 cells. Furthermore, in rats with dimethylnitrosamine （DMN）- induced hepatic fibrosis, MD-1 slowed the development and progression of hepatic fibrosis, protecting hepatic parenchymal cells and improving hepatic functions. Therefore, MD-1 is a potential drug for anti-hepatic fibrosis.展开更多
High mobility group box 1 protein （HMGB1） is a highly conserved, ubiquitous protein in the nuclei and cytoplasm of nearly all cell types. HMGB1 is secreted into the extracellular milieu and acts as a proinflammatory...High mobility group box 1 protein （HMGB1） is a highly conserved, ubiquitous protein in the nuclei and cytoplasm of nearly all cell types. HMGB1 is secreted into the extracellular milieu and acts as a proinflammatory cytokine. In this article we reviewed briefly the cellular immune response mediated by HMGB1 in inflammation and sepsis. This systemic review is mainly based on our own work and other related reports HMGB1 can actively affect the immune functions of many types of cells including T lymphocytes, regulatory T cells （Tregs）, dendritic cells （DCs）, macrophages, and natural killer cells （NK cells）. Various cellular responses can be mediated by HMGB1 which binds to cell-surface receptors [e.g., the receptor for advanced glycation end products （RAGE）, Toll-like receptor （TLR）2, and TLR4]. Anti-HMGB1 treatment, such as anti-HMGB1 polyclonal or monoclonal antibodies, inhibitors （e.g., ethyl pyruvate） and antagonists （e.g., A box）, can protect against sepsis lethality and give a wider window for the treatment opportunity. HMGB1 is an attractive target for the development of new therapeutic strategies in the treatment of patients with septic complications.展开更多
AIM： To investigate dynamic changes and significance of expression of NF-κBp65 in pancreatic tissues of rats with severe acute pancreatitis （SAP）, as well as BN52021 effects. METHODS： Wistar male rats were random...AIM： To investigate dynamic changes and significance of expression of NF-κBp65 in pancreatic tissues of rats with severe acute pancreatitis （SAP）, as well as BN52021 effects. METHODS： Wistar male rats were randomly divided into negative control group （NC group, n = 60）, SAP-model group （SAP group, n = 60）, and BN52021-treated group （BN group, n = 60）, and each of the above groups was respectively divided into 6 subgroups at different time points after operation （1 h, 2 h, 3 h, 6 h, 12 h, and 24 h） （n = 10）. By RT-PCR and Western blot, NF-κBp65 mRNA and its protein expression in pancreatic tissues of rats were detected respectively. RESULTS： The expression of NF-κBp65 mRNA dynamically changed in both SAP groups and BN groups. The mRNA level was higher in SAP groups than NC groups at 2 h, 3 h, 12 h, and 24 h after operation （P 〈 0.05）, higher in BN groups than NC groups at all time points （P 〈 0.05）, and higher in BN groups than SAP group at 1 h （P 〈 0.05）. The NF-κBp65 protein level was higher in SAP groups than NC groups at 1 h, 3 h, and 6 h （P 〈 0.01）, and 2 h, 12 h, and 24 h （P 〈 0.05）, higher in BN groups than NC groups at all time points （P 〈 0.05）, and lower in BN groups than SAP groups at 1 h, 3 h, and 6 h （P 〈 0.05）. CONCLUSION： The expression of NF-κBp65 in pancreatic tissues is dynamically changed and the changes play an important role in pathogenesis of SAR BN52021 exerts therapeutic effects through reducing the expression level of NF-κBp65 protein in the early stage of SAR展开更多
Perception of the phytohormone ethylene is accomplished by a small family of integral membrane receptors. In Arabidopsis, five ethylene receptor proteins are known, including ethylene resistant I （ETR1）. The hydroph...Perception of the phytohormone ethylene is accomplished by a small family of integral membrane receptors. In Arabidopsis, five ethylene receptor proteins are known, including ethylene resistant I （ETR1）. The hydrophobic aminoterminal domain of these receptors contains the ethylene-binding site while the carboxyl-terminal part consists of a histidine kinase domain and a response regulator domain, which are well known elements found in bacterial two-component signaling. The soluble membrane-extrinsic carboxyl-terminal part of the receptor, which is likely to play an important role in signal transduction, showed intrinsic kinase activity when expressed and purified on its own. However, a correlation between signal input and autokinase activity was not established in these studies, as receptors were missing the trans- membrane amino-terminal sensor domain. Thus, it is still unclear whether autophosphorylation occurs in response to perception of the ethylene signal. Here, we report on autophosphorylation studies of purified full-length ETR1. Autoki- nase activity of the purified receptor is controlled by ethylene or by ethylene agonists like the π-acceptor compound cyanide. In fact, both signal molecules were able to completely turn off the intrinsic kinase activity, Furthermore, the observed inhibition of autophosphorylation in ETR1 by both molecules could be prevented when the ethylene antagonist 1-methyl-cyclopropene （MOP） was applied.展开更多
Phospholipase A enzymes cleave phospho- and galactolipids to generate free fatty acids and lysolipids that function in animal and plant hormone signaling. Here, we describe three Arabidopsis patatin-related phospholip...Phospholipase A enzymes cleave phospho- and galactolipids to generate free fatty acids and lysolipids that function in animal and plant hormone signaling. Here, we describe three Arabidopsis patatin-related phospholipase A （pPLA） genes AtPLAIVA, AtPLAIVB, and AtPLAIVC and their corresponding proteins. Loss-of-function mutants reveal roles for these pPLAs in roots during normal development and under phosphate deprivation. AtPLAIVA is expressed strongly and exclusively in roots and AtplalVA-null mutants have reduced lateral root development, characteristic of an impaired auxin response. By contrast, AtPLAIVB is expressed weakly in roots, cotyledons, and leaves but is transcriptionally induced by auxin, although AtplalVB mutants develop normally. AtPLAIVC is expressed in the floral gynaecium and is induced by abscisic acid （ABA） or phosphate deficiency in roots. While an AtplalVC-1 loss-of-function mutant displays ABA respon- siveness, it exhibits an impaired response to phosphate deficiency during root development. Recombinant AtPLA proteins hydrolyze preferentially galactolipids and, less efficiently, phospholipids, although these enzymes are not localized in chloroplasts. We find that AtPLAIVA and AtPLAIVB are phosphorylated by calcium-dependent protein kinases in vitro and this enhances their activities on phosphatidylcholine but not on phosphatidylglycerol. Taken together, the data reveal novel functions of pPLAs in root development with individual roles at the interface between phosphate deficiency and auxin signaling.展开更多
OBJECTIVE: To investigate the inhibitory effect of Jinghua Weikang capsule(JWC) on gastric inflammation induced by Helicobacter pylori(H. pylori)via the nuclear factor-kappa B(NF-κB) signaling pathway in Kunming mice...OBJECTIVE: To investigate the inhibitory effect of Jinghua Weikang capsule(JWC) on gastric inflammation induced by Helicobacter pylori(H. pylori)via the nuclear factor-kappa B(NF-κB) signaling pathway in Kunming mice.METHODS: We investigated the anti-inflammation potential of JWC extract in vivo in a H. pylori-induced gastritis mouse model. The expression of inflammation-related molecules was evaluated by Western blotting, and the concentrations of in vivo inflammatory markers were measured by enzyme-linked immunosorbent assay. Inflammatory cell infiltration was evaluated by histopathological examination, and m RNA levels of related genes were evaluated by quantitative reverse transcription polymerase chain reaction.RESULTS: JWC had a dose-dependent protective effect against H. pylori-induced gastritis by protecting gastric epithelial cells and inhibiting inflammatory cell infiltration. Mechanistically, JWC decreased the protein levels of phosphorylated IκBαand NF-κB p65, m RNA levels of NF-κB pathway molecules, and plasma levels of tumor necrosis factor-α and interleukin 1 beta.CONCLUSION: An important finding of our study is that JWC attenuated gastrointestinal inflammation and ulceration and exerted a protective effect against gastric injury via inhibition of inflammation reactions and regulating the canonical NF-κB signaling pathway in vivo.展开更多
OBJECTIVE: To observe the impact of Xinfeng capsule(XFC) on cardiovascular function in adjuvant arthritis(AA) model rats and investigate the mechanism though toll-like receptor 4(TLR4)/nuclear factor kappa B(NF-κB) s...OBJECTIVE: To observe the impact of Xinfeng capsule(XFC) on cardiovascular function in adjuvant arthritis(AA) model rats and investigate the mechanism though toll-like receptor 4(TLR4)/nuclear factor kappa B(NF-κB) signaling pathway.METHODS: Seventy rats were randomly divided into seven groups: normal control(NC), model control(MC), tripterygium glycosides tablet(TPT),methotrexate(MTX), high, moderate and low dose XFC group. The administration began from day 19 after modeling for 30 day. Paw swelling, arthritic in-dex(AI), cardiac function indexes and myocardial pathological pattern were detected. The expression of TLR4, myeloid differentiation factor(My D) 88, interleukin-1 receptor-associated kinase(IRAK) 1, tumor necrosis factor receptor associated factor(TRAF) 6, NF-κB, tumor necrosis factor-alpha(TNF-α) proteins in myocardial tissue were determined by western blot method.RESULTS: Paw swelling and AI in MC group increased in MC group(P < 0.01), and decreased in high and moderate dose XFC groups(P < 0.01 or P > 0.05). Left ventricular systolic pressure(LVSP),left ventricular end-diastolic pressure(LVEDP),heart rate(HR) were elevated in MC group(P <0.01), and ± dp/dtmax and CI were reduced(P <0.01); while LVSP, LVEDP and HR declined and ±dp/dtmax, CI improved in high dose XFC group(P <0.05 or P < 0.01). LVSP in high dose XFC group were reduced more than other treatment groups(P <0.05 or P < 0.01). The improvements on LVEDP, dp/dt-max were superior to MTX and low dose XFC group, and the improvement on CI was better than low dose XFC group(P < 0.05 or P < 0.01). Myocardial fibers arranged irregular in MC group with intracellular edema and mitochondria damage. The modifications on myocardial structural were shown in each treatment group, but more prominent in TPT, high and moderate dose XFC group. The proteins of TLR4, My D88, IRAK1, TRAF6, NF-κB, TNF-αwere highly expressed in MC group, and those proteins declined in high and moderate dose XFC group(P < 0.05 or P < 0.01). High dose XFC group was superior to展开更多
文摘Matrine （MT）, the effective component of Sophora fla- vescens Air, has been shown to have anti-inflammation, immune-suppressive, anti-tumor, and anti-hepatic fibrosis activities. However, the pharmacological effects of MT still need to be strengthened due to its relatively low efficacy and short half-life. In the present study, we report a more effective thio derivative of MT, MD-1, and its inhibitory effects on the activation of hepatic stellate cells （HSCs） in both cell culture and animal models. Cytological experiments showed that MD-1 can inhibit the proliferation of HSC-T6 cells with a half-maximal inhibitory concentration （ICs0） of 62 pmollL. In addition, MD-1 more strongly inhibits the migration of HSC-T6 cells compared to MT and can more effectively induce G0/G1 arrest and apoptosis. Investigating the biological mechanisms underlying anti-hepatic fibrosis in the presence of MD-I, we found that MD-I can bind the epidermal growth factor receptor （EGFR） on the surface of HSC-T6 cells, which can further inhibit the phospho- rylaUon of EGFR and its downstream protein kinase B （Akt）, resulting in decreased expression of cyclin D1 and eventual inhibition of the activation of HSC-T6 cells. Furthermore, in rats with dimethylnitrosamine （DMN）- induced hepatic fibrosis, MD-1 slowed the development and progression of hepatic fibrosis, protecting hepatic parenchymal cells and improving hepatic functions. Therefore, MD-1 is a potential drug for anti-hepatic fibrosis.
基金supported,in part,by grants from the National Natural Science Foundation(Nos.81130035,30901561,30971192,81071545)the National Basic Research Program of China(No.2012CB518102)the China Postdoctoral Science Foundation(Nos.20100480347,201104125)
文摘High mobility group box 1 protein （HMGB1） is a highly conserved, ubiquitous protein in the nuclei and cytoplasm of nearly all cell types. HMGB1 is secreted into the extracellular milieu and acts as a proinflammatory cytokine. In this article we reviewed briefly the cellular immune response mediated by HMGB1 in inflammation and sepsis. This systemic review is mainly based on our own work and other related reports HMGB1 can actively affect the immune functions of many types of cells including T lymphocytes, regulatory T cells （Tregs）, dendritic cells （DCs）, macrophages, and natural killer cells （NK cells）. Various cellular responses can be mediated by HMGB1 which binds to cell-surface receptors [e.g., the receptor for advanced glycation end products （RAGE）, Toll-like receptor （TLR）2, and TLR4]. Anti-HMGB1 treatment, such as anti-HMGB1 polyclonal or monoclonal antibodies, inhibitors （e.g., ethyl pyruvate） and antagonists （e.g., A box）, can protect against sepsis lethality and give a wider window for the treatment opportunity. HMGB1 is an attractive target for the development of new therapeutic strategies in the treatment of patients with septic complications.
基金the National Natural Science Foundation of China, No. 30300465
文摘AIM： To investigate dynamic changes and significance of expression of NF-κBp65 in pancreatic tissues of rats with severe acute pancreatitis （SAP）, as well as BN52021 effects. METHODS： Wistar male rats were randomly divided into negative control group （NC group, n = 60）, SAP-model group （SAP group, n = 60）, and BN52021-treated group （BN group, n = 60）, and each of the above groups was respectively divided into 6 subgroups at different time points after operation （1 h, 2 h, 3 h, 6 h, 12 h, and 24 h） （n = 10）. By RT-PCR and Western blot, NF-κBp65 mRNA and its protein expression in pancreatic tissues of rats were detected respectively. RESULTS： The expression of NF-κBp65 mRNA dynamically changed in both SAP groups and BN groups. The mRNA level was higher in SAP groups than NC groups at 2 h, 3 h, 12 h, and 24 h after operation （P 〈 0.05）, higher in BN groups than NC groups at all time points （P 〈 0.05）, and higher in BN groups than SAP group at 1 h （P 〈 0.05）. The NF-κBp65 protein level was higher in SAP groups than NC groups at 1 h, 3 h, and 6 h （P 〈 0.01）, and 2 h, 12 h, and 24 h （P 〈 0.05）, higher in BN groups than NC groups at all time points （P 〈 0.05）, and lower in BN groups than SAP groups at 1 h, 3 h, and 6 h （P 〈 0.05）. CONCLUSION： The expression of NF-κBp65 in pancreatic tissues is dynamically changed and the changes play an important role in pathogenesis of SAR BN52021 exerts therapeutic effects through reducing the expression level of NF-κBp65 protein in the early stage of SAR
文摘Perception of the phytohormone ethylene is accomplished by a small family of integral membrane receptors. In Arabidopsis, five ethylene receptor proteins are known, including ethylene resistant I （ETR1）. The hydrophobic aminoterminal domain of these receptors contains the ethylene-binding site while the carboxyl-terminal part consists of a histidine kinase domain and a response regulator domain, which are well known elements found in bacterial two-component signaling. The soluble membrane-extrinsic carboxyl-terminal part of the receptor, which is likely to play an important role in signal transduction, showed intrinsic kinase activity when expressed and purified on its own. However, a correlation between signal input and autokinase activity was not established in these studies, as receptors were missing the trans- membrane amino-terminal sensor domain. Thus, it is still unclear whether autophosphorylation occurs in response to perception of the ethylene signal. Here, we report on autophosphorylation studies of purified full-length ETR1. Autoki- nase activity of the purified receptor is controlled by ethylene or by ethylene agonists like the π-acceptor compound cyanide. In fact, both signal molecules were able to completely turn off the intrinsic kinase activity, Furthermore, the observed inhibition of autophosphorylation in ETR1 by both molecules could be prevented when the ethylene antagonist 1-methyl-cyclopropene （MOP） was applied.
文摘Phospholipase A enzymes cleave phospho- and galactolipids to generate free fatty acids and lysolipids that function in animal and plant hormone signaling. Here, we describe three Arabidopsis patatin-related phospholipase A （pPLA） genes AtPLAIVA, AtPLAIVB, and AtPLAIVC and their corresponding proteins. Loss-of-function mutants reveal roles for these pPLAs in roots during normal development and under phosphate deprivation. AtPLAIVA is expressed strongly and exclusively in roots and AtplalVA-null mutants have reduced lateral root development, characteristic of an impaired auxin response. By contrast, AtPLAIVB is expressed weakly in roots, cotyledons, and leaves but is transcriptionally induced by auxin, although AtplalVB mutants develop normally. AtPLAIVC is expressed in the floral gynaecium and is induced by abscisic acid （ABA） or phosphate deficiency in roots. While an AtplalVC-1 loss-of-function mutant displays ABA respon- siveness, it exhibits an impaired response to phosphate deficiency during root development. Recombinant AtPLA proteins hydrolyze preferentially galactolipids and, less efficiently, phospholipids, although these enzymes are not localized in chloroplasts. We find that AtPLAIVA and AtPLAIVB are phosphorylated by calcium-dependent protein kinases in vitro and this enhances their activities on phosphatidylcholine but not on phosphatidylglycerol. Taken together, the data reveal novel functions of pPLAs in root development with individual roles at the interface between phosphate deficiency and auxin signaling.
基金Supported by Grants from the National Natural Science Foundation of China Project:Research on the Anti-inflammatory Effects and Mechanism of Jinghua Weikang Capsule and Its Ingredients Against H.pylori Infected Gastritis through TLR4-NFKB-cytokine Pathway in Mice(No.81473474)Natural Science Foundation of Beijing Project:the Effectand Mechanism of Jinghua Weikang Capsule Inhibiting the Adhesion Effect of H.pylori(No.7172220)
文摘OBJECTIVE: To investigate the inhibitory effect of Jinghua Weikang capsule(JWC) on gastric inflammation induced by Helicobacter pylori(H. pylori)via the nuclear factor-kappa B(NF-κB) signaling pathway in Kunming mice.METHODS: We investigated the anti-inflammation potential of JWC extract in vivo in a H. pylori-induced gastritis mouse model. The expression of inflammation-related molecules was evaluated by Western blotting, and the concentrations of in vivo inflammatory markers were measured by enzyme-linked immunosorbent assay. Inflammatory cell infiltration was evaluated by histopathological examination, and m RNA levels of related genes were evaluated by quantitative reverse transcription polymerase chain reaction.RESULTS: JWC had a dose-dependent protective effect against H. pylori-induced gastritis by protecting gastric epithelial cells and inhibiting inflammatory cell infiltration. Mechanistically, JWC decreased the protein levels of phosphorylated IκBαand NF-κB p65, m RNA levels of NF-κB pathway molecules, and plasma levels of tumor necrosis factor-α and interleukin 1 beta.CONCLUSION: An important finding of our study is that JWC attenuated gastrointestinal inflammation and ulceration and exerted a protective effect against gastric injury via inhibition of inflammation reactions and regulating the canonical NF-κB signaling pathway in vivo.
基金the National Natural Science Foundation of China:Research the Immune Mechanism of Xinfeng Capsule Treat the Heart Disease of AA Rats based on the mi R146a-TLR4/NF-κB Signal Pathway(No.81302967)The Key Subject Constructing Units by the State Administrative Bureau(No.[2009]30)+1 种基金Anhui Provincial Laboratory Construction Projects on Chinese Medicine Research and Development(No.[2008]150)Anhui Provincial Innovation Team in the Eleventh Five-Year Plan Period:Research and Development on Xin'an Medicine(2010TD005)
文摘OBJECTIVE: To observe the impact of Xinfeng capsule(XFC) on cardiovascular function in adjuvant arthritis(AA) model rats and investigate the mechanism though toll-like receptor 4(TLR4)/nuclear factor kappa B(NF-κB) signaling pathway.METHODS: Seventy rats were randomly divided into seven groups: normal control(NC), model control(MC), tripterygium glycosides tablet(TPT),methotrexate(MTX), high, moderate and low dose XFC group. The administration began from day 19 after modeling for 30 day. Paw swelling, arthritic in-dex(AI), cardiac function indexes and myocardial pathological pattern were detected. The expression of TLR4, myeloid differentiation factor(My D) 88, interleukin-1 receptor-associated kinase(IRAK) 1, tumor necrosis factor receptor associated factor(TRAF) 6, NF-κB, tumor necrosis factor-alpha(TNF-α) proteins in myocardial tissue were determined by western blot method.RESULTS: Paw swelling and AI in MC group increased in MC group(P < 0.01), and decreased in high and moderate dose XFC groups(P < 0.01 or P > 0.05). Left ventricular systolic pressure(LVSP),left ventricular end-diastolic pressure(LVEDP),heart rate(HR) were elevated in MC group(P <0.01), and ± dp/dtmax and CI were reduced(P <0.01); while LVSP, LVEDP and HR declined and ±dp/dtmax, CI improved in high dose XFC group(P <0.05 or P < 0.01). LVSP in high dose XFC group were reduced more than other treatment groups(P <0.05 or P < 0.01). The improvements on LVEDP, dp/dt-max were superior to MTX and low dose XFC group, and the improvement on CI was better than low dose XFC group(P < 0.05 or P < 0.01). Myocardial fibers arranged irregular in MC group with intracellular edema and mitochondria damage. The modifications on myocardial structural were shown in each treatment group, but more prominent in TPT, high and moderate dose XFC group. The proteins of TLR4, My D88, IRAK1, TRAF6, NF-κB, TNF-αwere highly expressed in MC group, and those proteins declined in high and moderate dose XFC group(P < 0.05 or P < 0.01). High dose XFC group was superior to
