SOM encodes a nucleus-localized CCCH-type zinc finger protein and negatively regulates seed germination in Arabidopsis thaliana. We have previously demonstrated that ectopic expression of Sl ABI3, an important transcr...SOM encodes a nucleus-localized CCCH-type zinc finger protein and negatively regulates seed germination in Arabidopsis thaliana. We have previously demonstrated that ectopic expression of Sl ABI3, an important transcription factor in abscisic acid（ABA） signaling pathway, resulted in alteration of Sl SOM expression patterns in both leaf and seed of tomato. In this study, we aimed to elucidate the function of tomato Sl SOM in regarding to seed germination and seedling development. Here, we constructed Sl SOM over-expression vector p BI121-SOM driven by Ca MV 35 S promoter, and the recombinant plasmid was incorporated into wild-type tomato by the method of Agrobacterium tumefaciens-mediated transformation. The result showed that over-expression of Sl SOM conferred enhanced responses to exogenous ABA application during seed germination and seedling development. In addition, ectopic expression of Sl SOM resulted in the alteration of expression level of ABA/GA（gibberellins） metabolic genes, such as Sl ABA1, Sl CYP707A1, Sl GA3ox2, and Sl GA2ox4, in both leaf and seed. The ABA anabolic gene Sl ABA1 and the GA catabolic gene Sl GA2ox4 were up-regulated while the ABA catabolic gene Sl CYP707A1 and the GA anabolic gene Sl GA3ox2 were down-regulated. Compared to wild type, the expression level of Sl ABI5 was increased by about 40–50% in transgenic seeds while adding exogenous ABA treatment. These results support the notion that Sl SOM inhibits seed germination by regulating ABA/GA metabolic genes and Sl ABI5 expression in Solanum lycopersicum.展开更多
Background:Amyloid-β(Aβ)metabolic imbalance is the pivotal pathogenesis leading to Alzheimer’s disease(AD).In sporadic AD,decreased clearance of Aβimportantly contributes to the onset and progression.Astrocytes,th...Background:Amyloid-β(Aβ)metabolic imbalance is the pivotal pathogenesis leading to Alzheimer’s disease(AD).In sporadic AD,decreased clearance of Aβimportantly contributes to the onset and progression.Astrocytes,the most abundant cell type in the brain,are mainly responsible for maintaining neuronal homeostasis.Most recently,it has been demonstrated that astrocytes play an important role in regulating Aβmetabolism.Icariin(ICA),a flavonoid glucoside extracted from the traditional Chinese herb Epimedium brevicornu,has been shown to produce protective effects against AD by decreasing Aβproduction.However,it remains unclear whether ICA regulates cellular Aβclearance in the astrocytes.Objective:To examine the regulatory effects of ICA on Aβremoval by astrocytes and explore the mechanisms of its actions.Methods:Uptake and subsequent degradation of Aβby astrocytes were evaluated using a combination of enzyme-linked immunosorbent assay(ELISA)and laser confocal microscopy.The effects of oligomer Aβ(oAβ1-42)and/or ICA on the expressions of sirt6 in the primary astrocytes were examined using western blotting and q-PCR assays.The expression of autophagy markers including P62 and LC3-Ⅱ,and phosphorylated-mTOR were measured by western blotting.In order to determine whether sirt6 is involved in the intracellular Aβmetabolism,sirt6 was knocked down using lentiviral vectors containing sirt6-shRNAs and autophagy levels were assessed by western blotting.Results:①In primary astrocytes,ICA not only significantly increased Aβinternalization but also obviously accelerated its degradation in a concentration-dependent manner.②Treatment of astrocytes with Aβ1-42 at 1μmol·L-1 significantly down-regulated the expression of sirt6,which was rescued by ICA.In addition,ICA at 20μmol·L-1 significantly increased the expression of LC3-Ⅱand markedly decreased the expression of P62 and phosphorylated-mTOR in primary astrocytes.③Sirt6 knockdown in primary astrocytes resulted in decreased cellular Aβuptake and degradation.Simultaneously,silencing of sirt6 in astrocytes increased P62 levels and reduced LC3-Ⅱand phosphorylated-mTOR levels.Conclusion:Taken together,our results demonstrate that sirt6 plays an important role in Aβmetabolism in astrocytes via induction of autophagy.ICA is a potential drug for treatment of AD as it upregulates cellular sirt6.展开更多
OBJECTIVE Microglia M1/M2 po⁃larization play pro-inflammatory and anti-inflam⁃matory roles,respectively,which is involved in memory decline.There is a close relationship between impaired baroreflex function and memory...OBJECTIVE Microglia M1/M2 po⁃larization play pro-inflammatory and anti-inflam⁃matory roles,respectively,which is involved in memory decline.There is a close relationship between impaired baroreflex function and memory impairment.The present study was designed to investigate whether arterial baroreflex deficiency induced by sinoaortic denervation(SAD)affected inflammation through modulation of M1/M2 polar⁃ization leading to the aggravation of learning and memory disorders in rats.METHODS Adult male SD rats were divided into four groups:the sham control,the SAD,the sham+scopolamine,the SAD+scopolamine.In another experiment,there were also four groups:the sham control,the SAD,the SAD+scopolamine and the SAD+scopolamine+ketanserin.All rats were examined for various behaviors using Morris water maze test,new object recognition test,and light dark shuttle test and Y maze test 4 weeks after sham or SAD surgery.CD16,CD206,IL-10,IL-6,IL-1βand TNF-αfrom hippocampus using Western blotting,immunofluorescence and turbidimetry.RESULTS Compared with the sham+scopol⁃amine,the SAD+scopolamine rats showed the reduced crossing times in Morris water maze test,the longer residence time in dark box during light dark shuttle test,and the decreased alterna⁃tion ratio in Y maze test.The level of CD206,IL-10,T-AOC and GSH was decreased,whereas CD16,IL-6,TNF-α,MDA was increased in the hippocampus of SAD+scopolamine rats.Addi⁃tionally,all the above changes were improved in the SAD+scopolamine+ketanserin rats when compared with the SAD+scopolamine.CONCLU⁃SION Arterial baroreflex dysfunction aggravates learning and memory disorders in rats,which may be related to the polarization of microglia.展开更多
OBJECTIVE Platelets play a major role in mediating inflammatory response.The present work was designed to investigate whether arterial baroreflex impairment induced by sinoaortic denervation(SAD)affect platelet activa...OBJECTIVE Platelets play a major role in mediating inflammatory response.The present work was designed to investigate whether arterial baroreflex impairment induced by sinoaortic denervation(SAD)affect platelet activation,leading to the exacerbation of cerebral cortex and hippocampus inflammation in rats.METHODS Adult male SD rats were divided into four groups:the sham control,the sinoaortic denervation(SAD),the sham+LPS,the SAD+LPS.In another experiment,there were also four groups:the sham control,the SAD,the SAD+LPS and the SAD+LPS+asprin.Four weeks after sham or SAD surgery,all rats were examined for the level of CD41,CD45,IL-1βand PF-4 in the cerebral cortex and hippocampus using immunofluorescence and ELISA.Blood platelet and leukocyte count,platelet microaggre⁃gation,expression of CD154 and CD62P on platelet surface and platelet-leukocyte aggregate level was detected by flow cytometry.RESULTS Compared with sham+LPS group,the in SAD+LPS group rats exhibited the high level of CD41,CD45,IL-1βand PF-4 in the cerebral cortex and hippocampus.Leukocyte count,platelet microag⁃gregation,expression of CD154 and CD62P on platelet surface and platelet-leukocyte aggregate level was increased,while blood platelet count was decreased in the SAD+LPS.Moreover,all the above changes were improved in the SAD+LPS+asprin group when compared with the SAD+LPS group.CONCLUSION Arterial baroreflex dysfunction exacerbates inflammation in the rat cerebral cortex and hippocampus,which is likely mediated by platelet.展开更多
OBJECTIVE To prepare gingerol dropping pills and to investigate its protective effect on alcoholic liver injury. METHODS The prescription was selected by orthogonal design method and the effect of the option and ratio...OBJECTIVE To prepare gingerol dropping pills and to investigate its protective effect on alcoholic liver injury. METHODS The prescription was selected by orthogonal design method and the effect of the option and ratio of ground substance,the temperature of drug. The hardness,circular degree,the tail formation and the dissolution time were studied. Totally 40 KM mice were randomly divided into control group,model group,gingerol dropping pill group(400 mg·kg^(-1)·d^(-1)) and positive control group(bifendate,150 mg·kg^(-1)·d^(-1)) of 10 mice each. The mice from the model and two drug groups were administrated with liqueur[0.15 mL/(10 g·d)]daily by gavage for 3 weeks,Two hours later,drug group mice were treated corresponding gingerol dropping pill and bifendate. Meanwhile,the control group were gavaged same amount of normal saline. Finally,when the model of acute alcoholic liver injury was established on the 22 stday,Biochemical indicators of ocular blood in mice were observed.We also observed the change of liver morphology. RESULTS Under optimum conditions,we can obtain dropping pills having circular shape,touching with hardness and short dissolution time. Compared with the control group,the levels of alanine transaminase(ALT),glutamic-oxaloacetic transaminase(AST) and malondialdehyde(MDA) in model group were obviously increased(P<0.01),While the activity of Superoxide dismutase(SOD) were decreased. In addition,In model group,mice liver disorders,hepatic lobule fusion,accompanying a large number of patchy sample liver cell vacuoles,various sizes of fat vacuoles appeared in cytoplasm and inflammatory cell infiltration were visible around the central vein. On the contrary,compared with the model group,drug groups attenuated or even reversed hepatic pathological changes. Form gingerol dropping pill group,an increase in hepatic SOD activity and serum ALT and AST activities were found and a significant decrease in hepatic MDA content were also observed(P<0.01). CONCLUSION The prescription of gingerol dropping pills was reasonable,and the preparation process was simple. Gingerol dropping pills can protect liver from alcoholic liver injury to some extend,and the mechanism may be related to its antioxidant effect.展开更多
Phosphodiesterase-4(PDE4)has been demonstrated to be a promising target for treatment of Alzheimer’s disease(AD).Roflumilast(Rof),a potent PDE4 inhibitor,has been approved for treatment of chronic obstructive pulmona...Phosphodiesterase-4(PDE4)has been demonstrated to be a promising target for treatment of Alzheimer’s disease(AD).Roflumilast(Rof),a potent PDE4 inhibitor,has been approved for treatment of chronic obstructive pulmonary disease(COPD)in humans.Recent studies have shown that Rof improves cognition at doses that do not cause an emetic response,the major side-effect of PDE4 inhibitors.However,the effect of Rof on cognition associated with AD remains largely unknown.Here we examined the effects of Rof on behavioral dysfunction and the related mechanisms in APP/PS1 double transgenic mice,a widely used model for AD.Mice at 10 months of age were first tested in novel object recognition for memory.The recognition index in APP/PS1 mice was decreased compared to WT mice,which was reversed by Rof at 5 and 10 mg·kg-1.This was then verified in the Morris water-maze test.The escape latency during acquisition training was significantly longer and the entries into the target quadrant during the probe trial were much less compared to WT controls,these were also reversed by Rof.In the tail-suspension and forced-swimming tests,which measure depression-like behavior,APP/PS1 mice showed prolonged immobility time,which was reversed by Rof.In addition,the staining of HE and Nissl showed that Rof reduced the loss of neurons and neurocyte apoptosis in APP/PS1 mice.It also reversed the decreased ratio of Bcl-2/BAX and inhibited the increased expression of PDE4D in the cerebral cortex and hippocampus of APP/PS1 mice.Finally,Rof reversed the decreased levels of cAMP and expression of phosphorylated cAMP response element-binding protein(CREB)and brain derived neurotrophic factor(BDNF)in APP/PS1 mice.Overall,these results suggest that Rof not only improves learning and memory,but attenuates depression-like behavior in AD mice,likely via PDE4D/cAMP/CREB/BDNF signaling-mediated neuroprotection.Therefore,Rof can be a therapeutic agent for AD,in particular the comorbidity of memory deficits and depression.展开更多
基金supported by the National Science Fund for Distinguished Young Scholars, China(30825030)the National Natural Science Foundation of China (31171179, 90717110)+1 种基金the National Basic Research Program of China (973, 2011CB100401)the Advanced Program of Doctoral Fund of Ministry of Education of China (20110181130009)
文摘SOM encodes a nucleus-localized CCCH-type zinc finger protein and negatively regulates seed germination in Arabidopsis thaliana. We have previously demonstrated that ectopic expression of Sl ABI3, an important transcription factor in abscisic acid（ABA） signaling pathway, resulted in alteration of Sl SOM expression patterns in both leaf and seed of tomato. In this study, we aimed to elucidate the function of tomato Sl SOM in regarding to seed germination and seedling development. Here, we constructed Sl SOM over-expression vector p BI121-SOM driven by Ca MV 35 S promoter, and the recombinant plasmid was incorporated into wild-type tomato by the method of Agrobacterium tumefaciens-mediated transformation. The result showed that over-expression of Sl SOM conferred enhanced responses to exogenous ABA application during seed germination and seedling development. In addition, ectopic expression of Sl SOM resulted in the alteration of expression level of ABA/GA（gibberellins） metabolic genes, such as Sl ABA1, Sl CYP707A1, Sl GA3ox2, and Sl GA2ox4, in both leaf and seed. The ABA anabolic gene Sl ABA1 and the GA catabolic gene Sl GA2ox4 were up-regulated while the ABA catabolic gene Sl CYP707A1 and the GA anabolic gene Sl GA3ox2 were down-regulated. Compared to wild type, the expression level of Sl ABI5 was increased by about 40–50% in transgenic seeds while adding exogenous ABA treatment. These results support the notion that Sl SOM inhibits seed germination by regulating ABA/GA metabolic genes and Sl ABI5 expression in Solanum lycopersicum.
文摘Background:Amyloid-β(Aβ)metabolic imbalance is the pivotal pathogenesis leading to Alzheimer’s disease(AD).In sporadic AD,decreased clearance of Aβimportantly contributes to the onset and progression.Astrocytes,the most abundant cell type in the brain,are mainly responsible for maintaining neuronal homeostasis.Most recently,it has been demonstrated that astrocytes play an important role in regulating Aβmetabolism.Icariin(ICA),a flavonoid glucoside extracted from the traditional Chinese herb Epimedium brevicornu,has been shown to produce protective effects against AD by decreasing Aβproduction.However,it remains unclear whether ICA regulates cellular Aβclearance in the astrocytes.Objective:To examine the regulatory effects of ICA on Aβremoval by astrocytes and explore the mechanisms of its actions.Methods:Uptake and subsequent degradation of Aβby astrocytes were evaluated using a combination of enzyme-linked immunosorbent assay(ELISA)and laser confocal microscopy.The effects of oligomer Aβ(oAβ1-42)and/or ICA on the expressions of sirt6 in the primary astrocytes were examined using western blotting and q-PCR assays.The expression of autophagy markers including P62 and LC3-Ⅱ,and phosphorylated-mTOR were measured by western blotting.In order to determine whether sirt6 is involved in the intracellular Aβmetabolism,sirt6 was knocked down using lentiviral vectors containing sirt6-shRNAs and autophagy levels were assessed by western blotting.Results:①In primary astrocytes,ICA not only significantly increased Aβinternalization but also obviously accelerated its degradation in a concentration-dependent manner.②Treatment of astrocytes with Aβ1-42 at 1μmol·L-1 significantly down-regulated the expression of sirt6,which was rescued by ICA.In addition,ICA at 20μmol·L-1 significantly increased the expression of LC3-Ⅱand markedly decreased the expression of P62 and phosphorylated-mTOR in primary astrocytes.③Sirt6 knockdown in primary astrocytes resulted in decreased cellular Aβuptake and degradation.Simultaneously,silencing of sirt6 in astrocytes increased P62 levels and reduced LC3-Ⅱand phosphorylated-mTOR levels.Conclusion:Taken together,our results demonstrate that sirt6 plays an important role in Aβmetabolism in astrocytes via induction of autophagy.ICA is a potential drug for treatment of AD as it upregulates cellular sirt6.
基金Natural Science Foundation of Shandong Province(ZR2017MH048)。
文摘OBJECTIVE Microglia M1/M2 po⁃larization play pro-inflammatory and anti-inflam⁃matory roles,respectively,which is involved in memory decline.There is a close relationship between impaired baroreflex function and memory impairment.The present study was designed to investigate whether arterial baroreflex deficiency induced by sinoaortic denervation(SAD)affected inflammation through modulation of M1/M2 polar⁃ization leading to the aggravation of learning and memory disorders in rats.METHODS Adult male SD rats were divided into four groups:the sham control,the SAD,the sham+scopolamine,the SAD+scopolamine.In another experiment,there were also four groups:the sham control,the SAD,the SAD+scopolamine and the SAD+scopolamine+ketanserin.All rats were examined for various behaviors using Morris water maze test,new object recognition test,and light dark shuttle test and Y maze test 4 weeks after sham or SAD surgery.CD16,CD206,IL-10,IL-6,IL-1βand TNF-αfrom hippocampus using Western blotting,immunofluorescence and turbidimetry.RESULTS Compared with the sham+scopol⁃amine,the SAD+scopolamine rats showed the reduced crossing times in Morris water maze test,the longer residence time in dark box during light dark shuttle test,and the decreased alterna⁃tion ratio in Y maze test.The level of CD206,IL-10,T-AOC and GSH was decreased,whereas CD16,IL-6,TNF-α,MDA was increased in the hippocampus of SAD+scopolamine rats.Addi⁃tionally,all the above changes were improved in the SAD+scopolamine+ketanserin rats when compared with the SAD+scopolamine.CONCLU⁃SION Arterial baroreflex dysfunction aggravates learning and memory disorders in rats,which may be related to the polarization of microglia.
基金Natural Science Foundation of Shandong Province(ZR2017MH048)。
文摘OBJECTIVE Platelets play a major role in mediating inflammatory response.The present work was designed to investigate whether arterial baroreflex impairment induced by sinoaortic denervation(SAD)affect platelet activation,leading to the exacerbation of cerebral cortex and hippocampus inflammation in rats.METHODS Adult male SD rats were divided into four groups:the sham control,the sinoaortic denervation(SAD),the sham+LPS,the SAD+LPS.In another experiment,there were also four groups:the sham control,the SAD,the SAD+LPS and the SAD+LPS+asprin.Four weeks after sham or SAD surgery,all rats were examined for the level of CD41,CD45,IL-1βand PF-4 in the cerebral cortex and hippocampus using immunofluorescence and ELISA.Blood platelet and leukocyte count,platelet microaggre⁃gation,expression of CD154 and CD62P on platelet surface and platelet-leukocyte aggregate level was detected by flow cytometry.RESULTS Compared with sham+LPS group,the in SAD+LPS group rats exhibited the high level of CD41,CD45,IL-1βand PF-4 in the cerebral cortex and hippocampus.Leukocyte count,platelet microag⁃gregation,expression of CD154 and CD62P on platelet surface and platelet-leukocyte aggregate level was increased,while blood platelet count was decreased in the SAD+LPS.Moreover,all the above changes were improved in the SAD+LPS+asprin group when compared with the SAD+LPS group.CONCLUSION Arterial baroreflex dysfunction exacerbates inflammation in the rat cerebral cortex and hippocampus,which is likely mediated by platelet.
基金The project supported by Col ege Students Of Science and Technology Innovation Project of Tai'an City(2015D064)the National College Students'Innovative and Entrepreneurial Training Project(201510439078)
文摘OBJECTIVE To prepare gingerol dropping pills and to investigate its protective effect on alcoholic liver injury. METHODS The prescription was selected by orthogonal design method and the effect of the option and ratio of ground substance,the temperature of drug. The hardness,circular degree,the tail formation and the dissolution time were studied. Totally 40 KM mice were randomly divided into control group,model group,gingerol dropping pill group(400 mg·kg^(-1)·d^(-1)) and positive control group(bifendate,150 mg·kg^(-1)·d^(-1)) of 10 mice each. The mice from the model and two drug groups were administrated with liqueur[0.15 mL/(10 g·d)]daily by gavage for 3 weeks,Two hours later,drug group mice were treated corresponding gingerol dropping pill and bifendate. Meanwhile,the control group were gavaged same amount of normal saline. Finally,when the model of acute alcoholic liver injury was established on the 22 stday,Biochemical indicators of ocular blood in mice were observed.We also observed the change of liver morphology. RESULTS Under optimum conditions,we can obtain dropping pills having circular shape,touching with hardness and short dissolution time. Compared with the control group,the levels of alanine transaminase(ALT),glutamic-oxaloacetic transaminase(AST) and malondialdehyde(MDA) in model group were obviously increased(P<0.01),While the activity of Superoxide dismutase(SOD) were decreased. In addition,In model group,mice liver disorders,hepatic lobule fusion,accompanying a large number of patchy sample liver cell vacuoles,various sizes of fat vacuoles appeared in cytoplasm and inflammatory cell infiltration were visible around the central vein. On the contrary,compared with the model group,drug groups attenuated or even reversed hepatic pathological changes. Form gingerol dropping pill group,an increase in hepatic SOD activity and serum ALT and AST activities were found and a significant decrease in hepatic MDA content were also observed(P<0.01). CONCLUSION The prescription of gingerol dropping pills was reasonable,and the preparation process was simple. Gingerol dropping pills can protect liver from alcoholic liver injury to some extend,and the mechanism may be related to its antioxidant effect.
基金This work was supported by research grants from National Natural Science Foundation of China(81773717 to HTZ81601229 and 81441111 to HW).
文摘Phosphodiesterase-4(PDE4)has been demonstrated to be a promising target for treatment of Alzheimer’s disease(AD).Roflumilast(Rof),a potent PDE4 inhibitor,has been approved for treatment of chronic obstructive pulmonary disease(COPD)in humans.Recent studies have shown that Rof improves cognition at doses that do not cause an emetic response,the major side-effect of PDE4 inhibitors.However,the effect of Rof on cognition associated with AD remains largely unknown.Here we examined the effects of Rof on behavioral dysfunction and the related mechanisms in APP/PS1 double transgenic mice,a widely used model for AD.Mice at 10 months of age were first tested in novel object recognition for memory.The recognition index in APP/PS1 mice was decreased compared to WT mice,which was reversed by Rof at 5 and 10 mg·kg-1.This was then verified in the Morris water-maze test.The escape latency during acquisition training was significantly longer and the entries into the target quadrant during the probe trial were much less compared to WT controls,these were also reversed by Rof.In the tail-suspension and forced-swimming tests,which measure depression-like behavior,APP/PS1 mice showed prolonged immobility time,which was reversed by Rof.In addition,the staining of HE and Nissl showed that Rof reduced the loss of neurons and neurocyte apoptosis in APP/PS1 mice.It also reversed the decreased ratio of Bcl-2/BAX and inhibited the increased expression of PDE4D in the cerebral cortex and hippocampus of APP/PS1 mice.Finally,Rof reversed the decreased levels of cAMP and expression of phosphorylated cAMP response element-binding protein(CREB)and brain derived neurotrophic factor(BDNF)in APP/PS1 mice.Overall,these results suggest that Rof not only improves learning and memory,but attenuates depression-like behavior in AD mice,likely via PDE4D/cAMP/CREB/BDNF signaling-mediated neuroprotection.Therefore,Rof can be a therapeutic agent for AD,in particular the comorbidity of memory deficits and depression.
