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ObjectiveTo investigate the mechanism of Dihuang Yinzi in improving astrocyte injury and protecting synaptic structure and function in the brain of Alzheimer's disease （AD） mice. MethodForty male APP/PS1 transgenic mice aged four months were randomly divided into a model group and a model + Dihuang Yinzi （0.25 g·kg-1） group， with 20 mice in each group. Forty C57BL/6J mice with the same background and same age were randomly divided into a control group and a control + Dihuang Yinzi （0.25 g·kg-1） group， with 20 mice in each group. The mice in the control + Dihuang Yinzi group and the model + Dihuang Yinzi group were administered with Dihuang Yinzi by gavage， and those in the control group and the model group received an equal volume of sterilized normal saline， once a day for 150 days. The learning and memory ability of mice was tested by the light-dark box test and Y-maze spontaneous alternation test. The content of glutamate （Glu） and glutamine （Gln） was measured by liquid chromatography-tandem mass spectrometry （LC-MS）. Long-term potentiation （LTP） assay was used to detect synaptic plasticity in brain tissues. The protein expression levels of excitatory amino acid transporter 2 （EAAT2）， postsynaptic density protein95 （PSD95）， and synaptophysin （SYN） in brain tissues were measured by Western blot. Immunofluorescence was used to assess the localization and expression of EAAT2. Colorimetry was performed to detect Na+-K+ ATPase activity in mouse brain tissues. ResultAs compared with the control group， the model group showed shortened residence latency （P<0.01）， increased number of errors （P<0.01） in the light-dark box test， reduced spontaneous alternation behaviors （P<0.01）， no significant difference in the total number of arm entries in the Y-maze spontaneous alternation test， down-regulated expression of EAAT2， PSD95， and SYN （P<0.01）， blunted activity of Na+-K+ ATPase （P<0.01）， up-regulated Glu level （P<0.01）， down-regulated Gln level （P<0.01）， and reduced relative population spike （PS） amplitude and the slope of excitatory postsynaptic potential （EPSP） （P<0.05， P<0.01）， while the above experimental indexes were not significantly different in the control + Dihuang Yinzi group. Compared with the model group， the model + Dihuang Yinzi group displayed prolonged residence latency （P<0.05）， decreased number of errors （P<0.01） in the light-dark box test， increased spontaneous alternation behaviors （P<0.01）， no significant difference in the total number of arm entries in the Y-maze spontaneous alternation test， up-regulated expression of EAAT2， PSD95， and SYN （P<0.01）， potentiated activity of Na+-K+ ATPase （P<0.01）， reduced Glu level （P<0.01）， up-regulated Gln level （P<0.01）， and increased PS amplitude and EPSP slope （P<0.01）. ConclusionDihuang Yinzi can improve cognitive dysfunction in AD mice by protecting astrocytes， increasing Glu uptake to reduce its abnormal accumulation， and protecting synaptic structure and function.
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ObjectiveTo explore the mechanism of Dihuang Yinzi （DHYZ）in improving astrocyte injury in the brain and regulating energy metabolism and autophagy disorder in Alzheimer's disease （AD） model mice. MethodForty male APP/PS1 transgenic mice aged four months were randomly divided into a model group and a model + DHYZ group （2.5 g·kg-1）， with 20 mice in each group. Forty C57BL/6J mice with the same background and same age were randomly divided into a control group and a control + DHYZ group （2.5 g·kg-1）， with 20 mice in each group. The mice in the control group and the model group were administered with an equal volume of sterilized normal saline by gavage， once a day for 150 days. Novel object recognition test and step-down test were performed to evaluate the learning and memory ability of mice. The expression of glial fibrillary acidic protein （GFAP） in astrocytes was detected by immunofluorescence and Western blot. High-performance liquid chromatography （HPLC） was used to detect adenosine triphosphate （ATP）， adenosine diphosphate （ADP）， and adenosine monophosphate （AMP） in brain tissues of mice， and the data obtained were used to calculate energy charge （EC） levels. The phosphorylation levels of liver kinase B1 （LKB1）， adenosine 5′-monophosphate （AMP）-activated protein kinase （AMPK）， UNC-51-like kinase 1 （ULK1）， and mammalian target of rapamycin （mTOR） and the expression levels of autophagy-related proteins Beclin-1， microtuble-associated protein 1 light chain 3 （LC3）-Ⅱ/LC3-Ⅰ， and p62 in mouse brain were measured by Western blot. ResultCompared with the control group， the model group showed decreased novel object recognition index， shortened retention latency， increased error times in the step-down test， up-regulated protein expression of GFAP， decreased content of ATP， ADP， and EC in brain tissues， elevated AMP ， increased levels of p-AMPK， p-LKB1， and p-mTOR， and protein expression of p62 ， and down-regulated p-ULK1 level and protein expression of Beclin-1 and LC3-Ⅱ/LC3-Ⅰ（P<0.01）， while the above experimental indexes were not significantly different in the control + DHYZ group. Compared with the model group， the model + DHYZ group showed increased novel object recognition index（P<0.05）， prolonged retention latency（P<0.01）， decreased error times（P<0.01） in the step-down test， reduced protein expression of GFAP（P<0.05）， increased content of ATP， ADP， and EC in brain tissues （P<0.05， P<0.01）， decreased AMP content（P<0.05）， reduced p-AMPK， p-LKB1， and p-mTOR levels and protein expression of p62， and up-regulated p-ULK1 level and protein expression of Beclin-1 and LC3-Ⅱ/LC3-Ⅰ（P<0.01）. ConclusionBy protecting astrocytes， DHYZ can improve energy metabolism and autophagy disorder in AD mice to improve the learning and memory ability of model mice.
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ObjectiveTo explore the mechanism of Dihuang Yinzi in improving astrocyte injury and glycolysis in Alzheimer's disease （AD） mice via regulating the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway， thereby improving the cognitive function of AD mice. MethodForty male APP/PS1 transgenic mice aged four months were randomly divided into a model group and a model + Dihuang Yinzi （0.25 g·kg-1） group， with 20 mice in each group. Forty C57BL/6J mice with the same background and same age were randomly divided into a control group and a control + Dihuang Yinzi （0.25 g·kg-1） group， with 20 mice in each group. The mice in the control + Dihuang Yinzi group and the model + Dihuang Yinzi group were administered with Dihuang Yinzi by gavage， and those in the control group and the model group received an equal volume of sterilized normal saline， once a day for 150 days. Morris water maze test was performed to test the ability of navigation and space exploration of mice. The protein expression of p-PI3K， PI3K， p-Akt， Akt， phosphofructokinase-1 （PFK-1）， and aldehyde dehydrogenase 3 family member B2 （ALDH3B2） in mouse brain tissues was measured by Western blot. An immunofluorescence assay was performed to detect astrocyte morphology and the expression level of ALDH3B2. ResultAs compared with the control group， the model group showed prolonged escape latency during the 2nd to 5th days of the location-based navigation （P<0.05， P<0.01）， reduced number of times crossing the target area of the platform， shortened residence time in the target quadrant （P<0.05， P<0.01）， prolonged residence time in the opposite quadrant （P<0.05）， increased surface area of the cell body and total length of cell protrusions of astrocytes （P<0.05， P<0.01）， and down-regulated protein expression of p-PI3K， p-Akt， ALDH3B2， and PFK-1 （P<0.01）， while the above experimental indexes were not significantly different in the control + Dihuang Yinzi group. Compared with the model group， the model + Dihuang Yinzi group showed shortened escape latency of APP/PS1 mice during the 2nd to 5th days of the location-based navigation （P<0.05， P<0.01）， increased number of times crossing the platform， prolonged target quadrant residence time （P<0.05， P<0.01）， shortened residence time in the opposite quadrant （P<0.05）， reduced surface area of the cell body and total length of cell protrusions of astrocytes （P<0.05）， and up-regulated protein expression of p-PI3K， p-Akt， ALDH3B2， and PFK-1 （P<0.01）. ConclusionDihuang Yinzi can improve the learning and memory ability of AD mice by activating the PI3K/Akt signaling pathway and up-regulating the protein expression of PFK-1 and ALDH3B2 to protect against astrocyte injury in brain tissues and improve glycolysis.
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Dysregulation of histone deacetylases (HDACs) is closely related to tumor development and progression. As promising anticancer targets, HDACs have gained a great deal of research interests and two decades of effort has led to the approval of five HDAC inhibitors (HDACis). However, currently traditional HDACis, although effective in approved indications, exhibit severe off-target toxicities and low sensitivities against solid tumors, which have urged the development of next-generation of HDACi. This review investigates the biological functions of HDACs, the roles of HDACs in oncogenesis, the structural features of different HDAC isoforms, isoform-selective inhibitors, combination therapies, multitarget agents and HDAC PROTACs. We hope these data could inspire readers with new ideas to develop novel HDACi with good isoform selectivity, efficient anticancer effect, attenuated adverse effect and reduced drug resistance.
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Objective:Based on network pharmacology to study the mechanism of Shaoyao-Gancao Decoction in treating tension-type headache. Methods:Searched for the active ingredients and potential targets of Shaoyao-Gancao Decoction from TCMSP database, and adopted the targets of tension-type headache from GeneCards, DisGeNET, Drugbank and OMIM databases. Then obtained all the intersections of Shaoyao-Gancao Decoction and tension-type headache, and uploaded them to the STRING databases to construct a PPI network and conduct topological properties analysis. Finally, established a Chinese medicine regulatory network of Chinese medicine-components-target genes-disease by Cytoscape 3.6.1 software. To perform the GO function enrichment analysis and KEGG analysis on the core targets. Results:There were 51 intersections of Shaoyao-Gancao Decoction and tension-type headache. The topological properties analysis suggested that CASP3, JUN, HSP90AA1, MAPK1, STAT3, CCND1, ESR1, RELA, PTGS2, MAPK14 may be the potential targets for the treatment of tension-type headache in Shaoyao-Gancao Decoction. Gene ontology enrichment analysis showed 876 biological processes, 101 molecular functions and 62 cellular components. KEGG pathway enrichment analysis showed 25 related signaling pathways, including TNF signaling pathway, serotonergic synapse, NOD-like receptor signaling pathway, Dopaminergic synapse and Sphingolipid signaling pathway. Conclusion:The treatment of tension-type headache by Shaoyao-Gancao Decoction verified the characteristics of multi-component, multi-target and multi-pathway, which provided reference for the clinical medication.
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Objective To study the mechanism of Liangruntongluo Recipe (Chinese medicines with functions of cooling, nourishing and dredging collaterals) and its modified formula in improving gastrointestinal function of diabete smellitus (DM) rats. Methods Male Wistar rats were used. Streptozotocin (STZ) was injected to rats to produce diabetic rat models. No Hypoglycemic drugs were administered to these rats to reduce blood glucose. After 18 weeks, warm water, Liangruntongino Recipe, Chinese medicines with function of nourishing, Chinese medicines with functions of cooling, and Cisapride were administered to the model rats. Detect the contents of plasma motilin (MOT), cholecystokinin (CCK), and somatostatin (SS) after 6 weeks. Results By affecting the secretion of gut hormone and having wide range of target, Liangrantongluo Recipe could regulate the disorder of gut hormone. The function of Liangruntongluo Recipe was better than its modified formula and cisapride.Conclusion Liangruntongluo Recipe can improve gastrointestinal dysfunction of DM rats.
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Objective To study the effects of eukaryotic expressive mature peptide LL-37 of human cationic antimicrobial peptide(hCAP-18) on the expressions of membrane molecules on dendritic cells(DCs).Methods By gene cloning,the eukaryotic expressive plasmid pcDNA4/Myc-His-LL-37 for the mature peptide LL-37 of hCAP-18 was constructed.Then they were transfected into HEK293 cell lines.After the cell lines were cultivated for 48 h,the supernatant was collected.Then the DCs from peripheral blood mononuclear cells(PBMCs) induced by rh-GM-CSF and rh-IL-4 were cultivated with the supernatant for 48h.The expressions of membrane molecules CD40 and HLA-DR on DCs were detected by flow cytometry(FCM).Results The eukaryotic expressive plasmid pcDNA4/Myc-His-LL-37 was constructed successfully and it expressed in eukaryotic cells HEK293.FCM results indicated that the expressions of CD40 and HLA-DR on the membrane of DCs which were stimulated by the supernatant produced by pcDNA4/Myc-His-LL-37 were higher than those in control group(P
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Objective To explore the dynamic alteration of intracellular free calcium concentration([ Ca2+]i),ATP level and membrane Ca2+-Mg2+ATPase acti vity of erythrocyte in the patients with acute cerebral infarction(CI).Methods we examined [Ca2+]i,ATP level and membrane Ca2+-Mg2+ATPase activity of erythrocyte in 30 patients with acute CI and 28 health controls by Fluorescence Activated Cell Sorter. Results [Ca2+]i in erythrocyte increased significantly in CI group(P<0.01),while the ATP level and membrane Ca2+-Mg 2+ATPase activity were lower than the controls(P<0.05,P<0.0 01).The above result was more remarkable in the elderly group than the young one .The dynamic alteration of [Ca2+]i in erythrocyte increased obv iously during 1～2 days after the attack,and reached the peak in 3～7 day s,it decreased slowly to the slightly low level at the beginning of th e attack in about two weeks,but it was still higher than the controls.The dynamic alteration of ATP level and Ca2+-Mg2+ATPase activit y after acute CI,it decreased significantly during 1～2 days after the at tack,and reached the lowest in 3～4 days.this status could last about one week.Then both of them increased slightly. There was remarkable negative correlation betwe en RBC [Ca2+]i and ATP level or membrance Ca2+-Mg2+ A TPase activity (r=-0.904,r=-0.978,P<0.05).There was positive correl ation between ATP lev el and membrane Ca2+-Mg2+ATPase activity(r=0.835,P<0.05 ).Conclusion There was calcium overload [Ca2+]i in th e intracellular of erythrocyte in acute CI,ATP level and membrane Ca2+ -Mg2+ATPase activity of erythrocyte CI was involved in the pathologi cal course of calcium overload,and related to the age.
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Objective To explore the dynamic alteration of intracellular free calcium concentration([Ca 2+ ]i),ATP level and membrane Ca 2+ Mg 2+ ATPase activity of erythrocyte in the patients with acute cerebral infarction(CI).Methods we examined [Ca 2+ ]i,ATP level and membrane Ca 2+ Mg 2+ ATPase activity of erythrocyte in 30 patients with acute CI and 28 health controls by Fluorescence Activated Cell Sorter.Results [Ca 2+ ]i in erythrocyte increased significantly in CI group( P