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Objective:To investigate the anti-anxious depression mechanism of Baihe Dihuangtang from the NOD-like receptor thermal protein domain 3 （NLRP3） inflammasome. Method:Fifty SD rats were randomly divided into normal group， model group， venlafaxine group （13.5 mg·kg<sup>-1</sup>）， Baihe Dihuangtang high and low dose group （16，4 g·kg<sup>-1</sup>）， with 10 rats in each group. Chronic restraint stress for 28 days （6 h） combined with subcutaneous injection of corticosterone （30 mg·kg<sup>-1</sup>） was used to establish induce an anxious depression model. From the 8th day of modeling， the rats in the normal group and the model group received distilled water， and those in groups with drug intervention were treated with corresponding drugs by gavage for 21 days. Elevated plus maze and open field test were used to evaluate the behavioral changes of rats. Enzyme- linked immunosorbent assay （ELISA） was used to detect serum and hippocampal interleukin-1<italic>β</italic> （IL-1<italic>β</italic>）， interleukin-6 （IL-6） and interleukin-18 （IL-18） levels. Western blot were used to detect the relative expression of hippocampal NLRP3， apoptosis-associated speck-like protein containing a CARD （ASC）， and Caspase-1. The pathological changes of the hippocampus were observed by hematoxylin-eosin（HE） staining， the average fluorescence intensity of NLRP3， ASC， and Caspase-1 was detected by immunofluorescence. The ultrastructure of neurons was observed under electron microscopy. Result:Compared with the normal group， the model group showed reduced total entries （TE）， the ratio of open-arm entries （OE%）， the ratio of open-arm times （OT%）， and the autonomous activity score （<italic>P</italic><0.01）， significant anxiety and depression-like behaviors， increased levels of IL-1<italic>β</italic>， IL-6， and IL-18 in the serum and hippocampus （<italic>P</italic><0.01）， elevated protein expression of NLRP3， ASC， and Caspase-1 （<italic>P</italic><0.01）， activated NLRP3 inflammasomes， and injured hippocampal neurons. Compared with the model group， the high-dose Baihe Dihuangtang group showed improved anxiety and depression-like behaviors （<italic>P</italic><0.01）， and decreased levels of IL-1<italic>β</italic>， IL-6， and IL-18 in the serum and hippocampus （<italic>P</italic><0.05，<italic>P</italic><0.01）， reduced protein expression of NLRP3， ASC， and Caspase-1 （<italic>P</italic><0.01）， and alleviated hippocampal neuron damage. Conclusion:Baihe Dihuangtang can improve neuronal damage in anxious depression by inhibiting the excessive activation of NLRP3 inflammasomes.
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To explore the effect of Baihe Dihuang Decoction on the synaptic plasticity of hippocampal neurons in rats with anxious depression. Fifty SD rats were randomly divided into normal group, model group, venlafaxine group(6.75 mg·kg~(-1)), high-dose Baihe Dihuang Decoction group(8.64 g·kg~(-1)) and low-dose Baihe Dihuang Decoction group(4.32 g·kg~(-1)). Chronic restraint stress(6 h) combined with corticosterone(ih, 30 mg·kg~(-1)) was used to establish an anxious depression model, and 7 days after modeling, the administration started and continued for 21 days. The anxiety and depression-like behaviors of the rats were evaluated. Golgi-Cox staining and electron microscopy were used to observe the morphology and ultrastructural changes of synaptic dendrites. Immunofluorescence was used to detect the expression of hippocampal synaptic plasticity protein synapsin-1 and postsynaptic density protein 95(PSD-95). Western blot method was used to detect the expression of functional protein synaptophysin(SYP) and synaptic Ras GTPase activating protein(SynGap). The results showed that the rats in the model group had obvious anxiety and depression-like behaviors, the hip-pocampal dendritic spine density and branch length were reduced, the number of synapses was cut, and the internal structure was da-maged. The average fluorescence intensity of synapsin-1 and PSD-95 was significantly reduced and the expression of SYP and SynGap also decreased. High-dose Baihe Dihuang Decoction could significantly improve the anxiety and depression-like behaviors of model rats, relieve synaptic damage, and increase the expression of synapsin-1, PSD-95, SYP, and SynGap proteins. Therefore, we believe that Baihe Dihuang Decoction can improve anxiety and depression behaviors by regulating the synaptic plasticity of hippocampal neurons.
Subject(s)
Animals , Rats , Depression/drug therapy , Hippocampus , Neuronal Plasticity , Rats, Sprague-Dawley , SynapsesABSTRACT
OBJECTIVE@#To study the effects of Zuogui Jiangtang Jieyu Formula (ZGJTJYF, the Chinese Medicine) on hippocampal neuron apoptosis in diabetes mellitus complicated with depression (DD).@*METHODS@#The primary cultured hippocampal neurons were treated with high glucose (150 mmol/L) and corticosterone (200 micromol/L) to establish the cell model of DD in vitro. The cultured hippocampal neurons were randomly divided into five groups: blank serum group, normal group, Zuogui Jiangtang Jieyu recipe drug-containing serum group, positive drug (metformin + fluoxetine) drug-containing serum group and model group (three compound holes in each group). The model group and the normal group were given the same amount of culture medium, and the other groups were given the corresponding serum with 10% volume fraction for 18 hours. Hoechst staining, high content cell imaging and RT-PCR were used to detect the apoptosis of hippocampal neurons and the expressions of apoptosis-related ETS-like 1 transcription factor(ELK-1), C-Jun N-terminal kinase(JNK) and c-Fos proteins and genes.@*RESULTS@#Compared with the blank group, the apoptotic number of hippocampal neurons in the model group was increased significantly, and the expression levels of ELK-1, JNK and c-Fos were increased significantly (P＜0.05). Compared with the model group, the local bright spots of hippocampal neurons in the Zuogui Jiangtang Jieyu recipe-containing serum group and the positive drug-containing serum group were decreased significantly, and the number of apoptotic cells was decreased significantly. The expressions of JNK, c-fos protein and mRNA were down-regulated significantly (P＜ 0.05), and the neural network and dendritic junction were improved significantly.@*CONCLUSION@#Zuo Gui Jiang Tang Jie Yu Formula can reverse the expressions of ELK-1, JNK and c-Fos signals in hippocampal neurons under DD environment and play an anti-apoptotic effect.
Subject(s)
Animals , Rats , Apoptosis , Depression , Drug Therapy , Diabetes Complications , Drug Therapy , Diabetes Mellitus , Drugs, Chinese Herbal , Pharmacology , Hippocampus , MAP Kinase Kinase 4 , Neurons , Proto-Oncogene Proteins c-fos , Random Allocation , ets-Domain Protein Elk-1ABSTRACT
Objective To investigate the effects of Xiaoyao Kang'ai Jieyu Formula on mice with breast cancer related depression; To explore its mechanism. Methods BALB/c female mice were set as study objects. Subaxillary injection of inflammatory 4T1 cells and intraperitoneal injection of corticosterone were used to establish a model of breast cancer related depression. According to the consumption of sugar water, mice were randomly divided into blank group, model group, paclitaxel group, Xiaoyao Kang'ai Jieyu Formula group, chemotherapy+fluoxetine group, and chemotherapy + Xiaoyao Kang'ai Jieyu Formula group. After 21 days, sucrose preference test, open-field test, and novelty feeding test were performed to detect changes in the behavior of mice in each group. ELISA were used to detected the contents of corticotrophin releasing hormone (CRH), adrenocorticotropic hormone (ACTH), and corticosterone (CORT) in plasma, and the contents of 5-hydroxytryptamine (5-HT), dopamine (DA), norepinephrine (NE), tumor markers carbohydrate antigen CA153, CA125 and carcinoembryonic antigen (CEA) in serum. Results Compared with the blank group, the consumption of sugar, horizontal and vertical movements of mice in the model group significantly decreased (P<0.01, P<0.05); the latency of ingestion significantly increased (P<0.01, P<0.05); the levels of tumor markers CA153, CA125, CEA and some indexes of HPA axis significantly increased (P<0.01); the expressions of monoamine neurotransmitter NE, DA and 5-HT significantly decreased (P<0.01). Compared with the model group, sugar consumption and autonomic activity significantly increased in the chemotherapy + Xiaoyao Kang'ai Jieyu Formula group, and behavioral despair condition was improved (P<0.01, P<0.05); the content of tumor markers and some indexes related to HPA axis significantly decreased (P<0.01, P<0.05); the contents of 5-HT, DA and NE significantly increased (P<0.01, P<0.05). Conclusion The combination of Xiaoyao Kang'ai Jieyu Formula and paclitaxel can effectively relieve the depressive symptoms of breast cancer related depression.
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Objective To observe the effects of Shuangdan Mingmu Capsules on the expressions of HIF-1α and NF-κB; To explore its possible mechanism of treatment. Methods SD rats were injected with STZ (50 mg/kg) into the tail vein to induce diabetes mellitus. After the DR model was confirmed by fundus fluorescein angiography, the rats were randomly divided into model group, Shuangdan Mingmu Capsules group and positive medicine group, and normal rats were set as control group. After the last administration, the blood glucose level and retinopathy in rats were measured. The protein and gene expression of HIF-1α and NF-κB in the retina were detected by Western blot and RT-PCR respectively. Results Compared with control group, the blood glucose in the model group was significantly increased; the retinal capillaries increased; the leakage of fluorescein was obvious; the protein and gene expressions of HIF-1α,NF-κB were significantly increased.After administration of Shuangdan Mingmu Capsules,the model rats decreased blood glucose; retinal microvascular volume decreased; fluorescein leakage area was significantly reduced; protein and gene expressions of HIF-1α and NF-κB were decreased. Conclusion Shuangdan Mingmu Capsules can effectively inhibit the neovascularization of DR rats and protect the retina, which may be related to the down-regulation of the expressions of HIF-1α and NF-κB.
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Objective To observe the effects of Zuogui Jiangtang Jieyu Prescription (ZGJTJYP) on pathological morphology of hippocampal neurons in rats of diabetes mellitus with depression (DD); To discuss its protective mechanism of action. Methods DD rat model was established by tail vein injection of STZ combined with chronic stress. Totally seventy-two SPF rats were randomly divided into normal group, model group, positive medicine group, ZGJTJYP low-, medium-, and high-dose groups, with 12 cases in each group, and were given corresponding medicine. After the last administration, blood glucose and behavioral changes were measured in each group. HE staining, Nissl staining, and Golgi staining were used to observe the changes of pathological morphology in rat hippocampus. The expression of Caspase-3 in hippocampus was detected by immunohistochemistry. Results Compared with the normal group, blood glucose level was significantly increased in the model group; the number of neurons in the hippocampus decreased in the model group; the apoptosis increased in the model group; the neuronal soma showed deformation and reduced dendritic spines and other pathological damages in the model group. After intervention of ZGJTJYP, hyperglycemia and depression-like status of the model rats were relieved; the number of hippocampal neurons increased significantly; neuronal morphology basically returned to normal. The expression of Caspase-3 in ZGJTJYP high-and medium-dose groups was significantly lower than that in model group. Conclusion ZGJTJYP can significantly improved blood glucose and learning and memory abilities in DD rats, which may be related to slowing hippocampal injury and apoptosis, and restoring neuronal quantity and morphology.
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<p><b>OBJECTIVE</b>To observe effect of acupuncture on serum macrophage inflammatory protein-2 (MIP-2) and MIP-2 mRNA expressions in isolated Fei and Dachang of severe acute pancreatitis (SAP) induced acute lung injury (ALI) rats in the acute phase.</p><p><b>METHODS</b>Forty male Wistar rats were randomly divided into four groups, i.e., the sham-operation group, the SAP group, the acupuncture treatment group, and the acupuncture control group, 10 in each group. The SAP model was induced by retrograde infusion of 3.5% sodium taurocholate into the pancreatobiliary duct. Under the guidance of "Fei and Dachang exterior-inferiorly related", points were acupunctured along Fei, Dachang, and Pi channels, as well as those points on the back of rats in the acupuncture treatment group 0.5 h after modeling. Besides, points were acupunctured along Fei and Pi channels, as well as those points on the back of rats in the acupuncture control group 0.5 h after modeling. Serum levels of tumor necrosis factor alpha (TNF-alpha) and nitric oxide (NO), and MIP-2 expressions were examined 6 h after modeling. Expressions of MIP-2 mRNA in isolated lung and large intestine tissues were detected by reverse transcription PCR.</p><p><b>RESULTS</b>Compared with the sham-operation group, serum levels of TNF-alpha and NO, and expressions of MIP-2 and MIP-2 mRNA in isolated lung and large intestine tissues were significantly higher in the SAP group (P < 0.05). Each index was lower in the acupuncture treatment group than in the SAP group and the acupuncture control group (P < 0.05). Besides, the serum level of MIP-2 and the MIP-2 mRNA expression in isolated lung and large intestine tissues were positively correlated in all groups except the sham-operation group (P < 0.05).</p><p><b>CONCLUSIONS</b>Under the guidance of "Fei and Dachang exterior-inferiorly related", acupuncture could remarkably reduce the severity of SAP induced ALI rats in the acute phase. Its mechanism might be related to suppressing over-expressions of MIP-2 mRNA in isolated lung and large intestine tissues, and lowering the serum MIP-2 expression level.</p>
Subject(s)
Animals , Male , Rats , Acupuncture Therapy , Acute Lung Injury , Blood , Metabolism , Chemokine CXCL2 , Blood , Genetics , Metabolism , Disease Models, Animal , Intestine, Large , Metabolism , Lung , Metabolism , Pancreatitis , Blood , Metabolism , RNA, Messenger , Genetics , Rats, WistarABSTRACT
Construction of mutant strain is an essential method in pathogenesis researches. The conventional method for Brucella unmarked deletion mutant construction is based on suicide plasmid, but the efficiency is very low. In the present study, we first optimized the electroporation parameters, and then, the cloning plasmid pEX18Gm containing sacB was successfully used to construct unmarked deletion mutant of the type IV secretion system. This indicated that by using conventional cloning plasmid as suicide plasmid in Brucella, unmarked deletion mutants can be constructed with high efficiency.