Effective time of immune suppressive protein of stress in serum

Immune suppressive protein of stress was generated in spleen and lymph node and released into serum in mice and rots under stress,which had strong suppressive effects on T- and B-lymphocyte proliferation.The present study showed that the serum from stressed mice presented a significant decrease in     

Suppressive effect of intestinal extract from stressed rats on lymphocyte proliferation

Our previous work showed that an immune suppressive protein, with a molecular weight of 155 and 370 kDa, was generated in spleen and lymph node in rats treated with stress. The protein strongly inhibited lymphocyte proliferation in vitro. In the present study the extract of intestine from the rats treated with restraint stress was also suppressive on lymphocyte proliferation.The further experiments showed that the bio-activity and molecular weight of the extract were all similar to the     

Supppressive effect of the immuno-suppressive protein induced by stress on arterrial blood pressure rat

To study the effect of partially purified immune suppressive protein of stress on the normal arterial blood pressure in rots.The immune suppressive protein of stress was partially purified from the lymph node of the restraint stressed rat with salting-out, extra-filtration and HPLC gel filtration.The protein was intravenously injected into normal SD rats.Blood pressure, heart rate and respiration were recorded with MacLab system.It was found that intravenous injection of the protein (200, 400, and 800 μg per     

The generation of IPSP was antagonized by injection of 6-OHDA

In the present study 6 hydroxydopamine (6-OHDA) was used for chemical sympathectomy and the effect of 6-OHDA on generation of immune suppressive protein of stress (ISPS) was studied.It was found that the generation of ISPS was significantly suppressed.The maximal suppression was found on d 5 after 6-OHDA injection and it recovered gradually to normal on d 14.In order to assess the effectiveness of the 6-OHDA injection, the contents of norepinephrine (NE) in spleen and lymph node were assayed by high performance liquid chromatography-electrochemical detector (HPLC-ED).NE was     

T-lymphocyte generates an immune suppressive protein induced by stress

Our previous work demonstrated that under the conditions of restraint stress and under the control of central nervous system (CNS), an immune suppressive protein of stress (ISPS) was generated in peripheral lymph tissue and released into the blood stream, which acts as an immune suppressor.In the present work the cellular source of ISPS was studied.Firstly,the extracts from various cell types were prepared and the identification was performed by bioassay of lymphocyte proliferation.The results showed that the extract of mononuclear     

Brain IL-1β was involved in reserpine-induced behavioral depression in rats

AIM: To investigate the mechanism of brain interleukin-1β(IL-1β) in reserpine-induced behavioral depression in rats. METHODS: Porsult swim test was used in the measurement of depressive behavior and ELISA was used in measurement of brain IL-1β. RESULTS: Intraperitoneal injection of reserpine (0, 4, 6, and 8 mg/kg, ip) increased floating time in the Porsult swim test in a dose-and time-dependent manner in rats. Intracerebroventricular injection(icv) of IL-1β receptor antagonist (IL-1ra, 6 mg/kg) blocked the increment of floating time in Porsult swim test at 48 and 72 h after reserpine injection, but not at 1 and 24 h after injection. Brain IL-Iβ increased after reserpine treatment in posterior cortex, hippocampus, and hypothalamus. The increase of IL-1β concentration starts at 24 hours after injection of reserpine and reached the peak at 48 h. CONCLUSION: Reserpine induced behavioral depression partially via brain interleukin-1β generation.     

Positive reaction of direct popliteal lymph node assay isn't induced in humanized NPG mice with reconstituted immune system北大核心CSCD

OBJECTIVE: To perform direct popliteal lymph node assay (d-PLNA) induced by D-penicillamine hydrochloride (D-pen) or streptozotocin (STZ) in humanized NPG (hu-NPG) mice with a reconstituted immune system, NPG mice and BALB/c mice respectively, and to compare the responses in these mice to elucidate the mechanism of d-PLNA. METHODS: Female NPG mice were irradiated and transplanted with human hematopoietic stem cells isolated from cord blood to form hu-NPG mice. The number and function of peripheral blood lymphocytes in hu-NPG mice were detected by flow cytometry and lymphocyte proliferation test. The hu-NPG, NPG and BALB/c mice were randomly divided into D-pen group and STZ group (5 mice per group), respectively. Then, the positive chemical D-pen (1 mg per mouse) or STZ (0.75 mg per mouse) was injected subcutaneously into the right hind footpad of these mice using the established d-PLNA protocol. On the 7th day after injection, the mice were sacrificed. The bilateral popliteal lymph nodes (PLNs) were weighed. The difference of d-PLNA response was evaluated at 7 d after the injection of positive drugs. RESULTS: All the NPG, hu-NPG and BALB/c mice behaved normally after the injection of positive drugs. No systemic toxicity was observed. The symptom of local irritation disappeared within 7 d. As in previous studies, the BALB/c mice showed a typical positive response of d-PLNA, as evidenced by the increase in both mass and cell count of the PLNs in the treated lateral (mass index >2 and cellularity index >5), while NPG mice and hu-NPG mice showed a negative resoponse. Pathological examinations demonstrated that the PLNs of NPG mice and hu-NPG mice had developmental defect in lymphoid tissue and were smaller in size than BALB/c mice. Phenotypic analysis of the peripheral blood lymphocytes in hu-NPG mice showed that the majority of human lymphocytes expressed B cell markers CD45+CD19+ (75.5±6.6)%, and only about (17.1±6.6)% of the lymphocytes expressed T cell markers CD45+CD3+. There were few lymphocytes in NPG mice. In addition, these cells did not proliferate with mitogen stimulation in the lymphocyte proliferation test. CONCLUSION: The d-PLNA reaction induced by D-pen or STZ in mice is closely related to the number and normal function of lymphocytes in vivo. Negative d-PLNA reaction results from a lack of normal lymphocytes in NPG mice or from defected human lymphocyte function in hu-NPG mice. Thus hu-NPG mice are currently not suitable for d-PLNA.     

Effects of Xanthoceraside on learning and memory impairment in mice induced by i.c.v.Aβ_(1-42)

Objective To investigate the improvement of Xanthoceraside on learning and memory impairment in mice induced by intracerebroventricular injection of Aβ1-42(i.c.v.Aβ1-42)and the possible mechanisms of its protection against AD.Methods Y-maze test,water-maze test and step-down test were used to investigate the learning and memory ability of mice;Biochemical analysis was used to detect the activity of CAT,T-AOC,ATPase and the content of MDA.Results The results showed that Xanthoceraside could significantly increase the alternation behavior in Y-maze test,shorten swimming time in water maze test and increase the latency and decrease the number of errors and the total time of shock in step-down test.Xanthoceraside markedly increased the activity of CAT,T-AOC,ATPase,at the same time,decreased the content of MDA.Conclusions Xanthoceraside can improve learning and memory impairment in mice induced by i.c.v.Aβ1-42 significantly.The mechanism may be associated with the protection against damage induced by free radicals;the inhibition of membrane lipid peroxidation and the improvement of metabolism of brain.     

Effect of telmisartan on expression of protein kinase C-α in kidneys of diabetic mice

Aim： To investigate the effects of angiotensin receptor blocker （ARB） telmisartan on the expression and distribution of protein kinase C （PKC）-α in the kidneys of diabetic mice. Methods： Diabetic mice were induced with streptozotocin and a group of them were randomly selected for treatment with telmisartan. After 6 weeks, the expression and localization of PKC-α in the renal cortex, and the outer and inner medulla were assessed by immunohistochemistry and semiquantitative Western blotting. In addition, expressions of PKC-α, transforming growth factor- β1 （TGF-β1）, and vascular endothelial growth factor （VEGF） in glomeruli were measured by semiquantitative immunohistochemistry. Results： Diabetic and normal mice showed similar distributions of PKC-α in the kidneys. The expression of PKC-α was found in glomeruli, epithelial cells of proximal tubules, and medullary- collecting duct, while not in the medullary and cortical thick ascending limb, and was different in the epithelial cells of proximal tubules of diabetic nephropathy （DN） mice, PKC-α was mostly translocated from the basement membrane to the apical membrane, whereas it was largely translocated from the apical membrane to the basement membrane in epithelial cells of the inner medullary-collecting duct. Western blotting detected increased expression of PKC-α in the renal cortex and outer medulla, but not in the inner medulla of DN mice. Enhanced expressions of PKC-α, TGF-β1, and VEGF were shown in the glomeruli of DN mice, where PKC-α exhibited a correlation to VEGF, but no correlation to TGF-β1. ARB telmisartan attenuated alterations of PKC-α as mentioned earlier in the DN mice. Conclusion： Our findings suggest that PKC-α may play a role in the pathogenesis of DN, and that the nephroprotective effects of ARB telmisartan may be partly associated with its influence on PKC-α.     

雷丸多糖的抗炎及免疫刺激作用

Lei Wan, Poliporus mylittae Cook et Mass(Omphalia lapidescena Scbraet) is a kind of fungus used in traditional Chinese medicine, as an antheiminthie From Lei Wan, an "active component designated as. S-4001 had been isolated.Preliminary results indicate that S-4001 belongs to D, β, 1-3 glucan with some 1-6 linkages.After administration of S-4001, significant antiinflammatory activity was found in various experimental animal models, including croton oil induced ear edema in mice and agar or yeast induced ankle swelling in rats. An inhibitory action on leucocyte migration inducced by intraperitoneal injection of CMC in rats was also observed. The plasma content of corticosterone was significantly increased, but the content of ascorbic acid in the adrenals did not change in rats given S-4001. Apart from these actions, S-4001 showed a number of immunostimulating actions such as increasing the clearance of Congo red from mice blood and potentiating the immunohemolysis reaction in 615 mice.     

Hydrogen sulfide inhibits homocysteine-induced endoplasmic reticulum stress and neuronal apoptosis in rat hippocampus via upregulation of the BDNF-TrkB pathway

Aim： Homocysteine （Hcy） can elicit neuronal cell death, and hyperhomocysteinemia is a strong independent risk factor for Alzheimer’s disease. The aim of this study was to examine the effects of hydrogen sulfide （H2S） on Hcy-induced endoplasmic reticulum （ER） stress and neuronal apoptosis in rat hippocampus.
Methods： Adult male SD rats were intracerebroventricularly （icv） injected with Hcy （0.6 μmol/d） for 7 d. Before Hcy injection, the rats were treated with NaHS （30 or 100 μmol·kg^-1·d^-1, ip） and/or k252a （1 μg/d, icv） for 2 d. The apoptotic neurons were detected in hippocampal coronal slices with TUNEL staining. The expression of glucose regulated protein 78 （GRP78）, C/EBP homologous protein （CHOP）, cleaved caspase-12, and BDNF in the hippocampus were examined using Western blotting assays. The generation of H2S in the hippocampus was measured with the NNDPD method.
Results： Hcy markedly inhibited the production of endogenous H2S and increased apoptotic neurons in the hippocampus. Further-more, Hcy induced ER stress responses in the hippocampus, as indicated by the upregulation of GRP78, CHOP, and cleaved caspase-12. Treatment with the H2S donor NaHS increased the endogenous H2S production and BDNF expression in a dose-dependent manner, and significantly reduced Hcy-induced neuronal apoptosis and ER stress responses in the hippocampus. Treatment with k252a, a specific inhibitor of TrkB （the receptor of BDNF）, abolished the protective effects of NaHS against Hcy-induced ER stress in the hippocampus.
Conclusion： H2S attenuates ER stress and neuronal apoptosis in the hippocampus of Hcy-treated rats via upregulating the BDNF-TrkB pathway.     

Potassium 2-（1-hydroxypentyl）-benzoate promotes long-term potentiation in Aβ1-42-injected rats and APP/PS1 transgenic mice

Aim： Potassium 2-（1-hydroxypentyl）-benzoate （d/-PHPB） is a new drug candidate for ischemic stroke. The aim of this study was to investigate the effects of dI-PHPB on memory deficits and long-term potentiation （LTP） impairment in animal models of Alzheimer＇s disease. Methods： The expression of NMDA receptor subunits GluN1 and GluN2B in the hippocampus and cortex of APP/PS1 transgenic mice were detected using Western blot analysis. Memory deficits of the mice were evaluated with the passive avoidance test. LTP impairment was studied in the dentate region of Aβ1-42-injected rats and APP/PS1 transgenic mice. Results： APP/PS1 transgenic mice showed significantly lower levels of GluN1 and p-GluN2B in hippocampus, and chronic administration of dI-PHPB （100 mg·kg-1·d1, po） reversed the downregulation of p-GluN2B, but did not change GluN1 level in the hippocampus. Furthermore, chronic administration of d/-PHPB reversed the memory deficits in APP/PS1 transgenic mice. In the dentate region of normal rats, injection of dI-PHPB （100 μmol/L, icv） did not change the basal synaptic transmission, but significantly enhanced the high-frequency stimulation （HFS）-induced LTP, which was completely prevented by pre-injection of APV （150 μmol/L, icv）. Chronic administration of dI-PHPB （100 mg·kg-1·d-1, po） reversed LTP impairment in Aβ1-42 -injected normal rats and APP/PS1 transgenic mice. Conclusion： Chronic administration of d/-PHPB improves learning and memory and promotes LTP in the animal models of Alzheimer＇s disease, possibly via increasing p-GluN2B expression in the hippocampus.     

Effect of matrine hydrochloride on liver injury

Objective Searching the function that the Injection of the matrine hydrochloride prevents and cures acute chemical liver injury of mice、immunity liver injury of mice and chronic liver injury of rats.Methods Acute hepatic injury models of mice induced by Chemical poison carbon tetrachloride(CCl4),thioacetamide(TAA),D-galactosamine(D-GalN),immunity hepatic injury model of mice induced by BCG and fat polysaccharide(LPS),chronic liver injury model of rats induced by CCl4 were introduced in the experiment.The serum ALT and AST were measured in acute hepatic injury experiments.Serum ALT,AST,AKP,ALB,TP,BiL-T,creatinine,triglyceride,sialic acid,laminin,hyaluronic acid,type Ⅲ procollagen and type Ⅳ collagen,hepatic hydroxyproline(HyP)of rats in chronic liver injury animals were determined after Injection of the matrine hydrochloride.Results The Injection of the matrine hydrochloride reduced serum ALT and AST level of acute chemical liver injury of mice induced by CCl4,TAA and D-GalN.The index of the liver and the spleen of immunity liver injury of mice induced by BCG and LPS were decreased after the injection of matrine hydrochloride treatment.Compared with the model group,the injection may obviously inhibited serum ALT,AST,TP,AKP,TRI,BiL-T,creatinine,triglyceride,sialic acid,laminin,hyaluronic acid,type Ⅲ procollagen and type Ⅳ collagen activity of chronic liver injury of rats induced by CCl4,elevated ALB、A/G,reduced the liver HyP,decreased the index of the liver and the spleen.The liver visual observation,the pathology inspection and the HAI grading result showed the injection may reduce the inflammatory activity in liver tissue,restrain the liver cell damage,reduce the pseudolobuli formation.Conclusions The Injection of matrine hydrochloride had the protective function to acute chemical hepatic injury of mice induced by CCl4、TAA、D-GalN、immunity hepatic injury of mice induced by the BCG and LPS and chronic liver injury of rats induced by CCl4.     

Effect of Sea Cucumber Enzymolysis Fermentation Liquid on immunosuppressed mice based on metabonomics北大核心CSCD

Aim To prepare the sea cucumber enzy¬molysis fermentation liquid (SCEFL) by enzymatic hydrolysis of protease and fermentation of probiotics and to investigate the effect of SCEFL on the immunosup-pression induced by cyclophosphamide in mice and to explore its mechanism by metabomic method. Methods The immunosuppressive model was induced by in-traperitoneal injection of cyclophosphamide. C57BL/6J mice were randomly divided into normal group, model group, Levamisole group, SCEFL groups (at low, medium and high doses). The pathological changes of spleen were observed by HE staining. The proportion of CD4+and CD8+T lymphocyte subsets and the pro-portion of CD4 /CD8 T lymphocyte subsets in peripheral blood were detected by flow cytometry. The con¬tents of IL-2 ( interleukin-2 ), IgG ( immunoglobulin G) and IgM (immunoglobulin G) in serum were detected by ELISA. The changes of endogenous metabolites in mouse serum were analyzed by UPLC-Q/TOF-MS metabolomics, and the related metabolic pathways were explored. Results SCEFL could improve the pathological changes of immunosuppressed mice, in¬crease the proportion of CD4 T cells and the proportion of CD4+/CD8+T lymphocyte subsets,and increase the contents of IL-2, IgG and IgM in serum. The immune function of mice was regulated mainly through six related metabolic pathways, including glyc-erophospholipid metabolism, sphingolipid metabolism, linoleic acid metabolism, a-linoleic acid metabolism, glycophosphatidylinositol-anchored biosynthesis and arachidonic acid metabolism. Conclusion SCEFL may ameliorate the immunosuppression of mice by regulating endogenous metabolic disorder. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

Gene expression profile of amyloid beta protein-injected mouse model for Alzheimer disease

Aim: To investigate the gene expression profile changes in the cerebral cortex of mice injected icv with amyloid beta-protein (Aβ) fragment 25-35 using cDNA microarray. Methods: Balb/c mice were randomly divided into a control group and Aβ-treated group. The Morris water maze test was performed to detect the effect of Aβ-injection on the learning and memory of mice. Atlas Mouse 1.2 Expression Arrays containing 1176 genes were used to investigate the gene expression pattern of each group. Results: The gene expression profiles showed that 19 genes including TBX1, NF-κB, AP-1/c-Jun, cadherin, integrin, erb-B2, and FGFR1 were up-regulated after 2 weeks of icv administration of Aβ; while 12 genes were down-regulated, including NGF, glucose phosphate isomerase 1, AT motifbindingfactor 1, Na^ /K^ -ATPase, and Akt. Conclusions: The results provide important leads for pursuing a more complete understanding of the molecular events of Aβ-injection into mice with Alzheimer disease.     

Suppression of complete Freund＇s adjuvant-induced adjuvant arthritis by cobratoxin

Aim： Cobratoxin （CTX）, the long-chain α-neurotoxin from Thailand cobra venom, has been demonstrated to have analgesic action in rodent pain models. The present study evaluated the anti-inflammatory and anti-nociceptive effects of CTX on adjuvant arthritis （AA） in rats. Methods： Arthritis was induced by injection of complete Freund＇s adjuvant （CFA） in rats. Paw swelling and hyperalgesia of AA rats were measured at various times after CFA administration. Tumor necrosis factor-a （TNF-α）, interleukin-1 （IL-1）, interleukin-2 （IL-2） and interleukin-10 （IL-10） levels in serum were determined with ELISA. Histopathological changes in synoviocytes were examined under a microscope. Involvement of the cholinergic system in the effects of CTX was examined by pretreatment of animals with the α7 nicotinic receptor （α7-nAChR） antagonist methyllycaconitine （MLA）. Results： CFA induced marked paw swelling and reduced thresholds of mechanical and cold-induced paw withdrawal. The levels of TNF-α, IL-1 and IL-2 in the serum of AA rats were increased, whereas the level of IL-10 was decreased. Histopathological examination of synoviocytes showed pronounced inflammation and accumulation of collagen. The administration of CTX （17.0 μg/kg, ip） significantly reduced paw swelling and mechanical and thermal hyperalgesia. CTX also reduced the production ofTNF-α, IL-1, and IL-2 but increased the production of IL-10 and altered pathohistological changes. The analgesic and anti-inflammatory efficacy of CTX was significantly reduced by MLA （3 mg/kg, sc）. Conclusion： These results indicate that CTX has a beneficial effect on CFA-induced arthritis by modulating the production of inflammatory cytokines, α7-nAChR appears to mediate the anti-nociceptive and anti-inflammatory actions of CTX.     

Effect of human urinary kallidinogenase on cognitive function of SAMP8 mouse model and its mechanism北大核心CSCD

Aim To study the effect of human urinary kallidinogenase(HUK)on the cognitive function of SAMP8 mouse model and its mechanism. Methods SAMP8 mice were divided intofive groups:SAMP8 group,treatment group(giving 8.75×10-3,1.75×10-2,3.5×10-2,7.0×10-2 HUK),and the SAMR1 vehicle group was used as blank control. Each group was performed Morris water maze to detect spatial cognition. Afterwards the group with the most obvious cognitive improvement(HUK group)was selected for the follow-up experiments. Immunohistochemical detection of ChAT expression in CA3 area was further verified by RtPCR. Western blot was used to detect the expression of PSD95,SYN,BDNF,and pCREB protein. The activity of MPO and the content of IL-1β and IL-18 were determined. Results The passing times in the SAMP8 group was less than that of the SAMR1 group(P<0.05). The passing times of treatment group increased compared with the SAMP8 group(P<0.05 or P<0.01),and the spatial probe time of the target quadrant was shorter(P<0.05 or P<0.01). We conducted follow-up experiments with group d(HUK group). The expression of ChAT positive cells in CA3 area of SAMP8 group was significantly lower than that of SAMR1 group; the expression of positive cells in HUK group significantly increased; RtPCR showed that ChAT expression in SAMP8 group was lower than that in SAMR1 group,and ChAT expression was significantly higher than that in SAMP8 group after HUK treatment. Compared with the SAMR1 group,the levels of IL-1β,IL-18 and MPO activity in the CA3 area of SAMP8 group significantly increased,and the protein expressions of PSD95,SYN,BNDF and pCREB decreased. After HUK treatment,the content of IL-1β,IL-18 and MPO activity decreased,and the expression of PSD95,SYN,BNDF and pCREB increased. Conclusions HUK can improve the spatial cognition of SAMP8 mice. The mechanism may be achieved by promoting the expression of ChAT in CA3 area,reducing the oxidative stress and increasing synapse-related proteins. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

Effects of melatonin on pethidine-induced physical dependence and its antioxidative action

Objective To study the effects of melatonin on pethidine-induced physical dependence and its antioxidative action.Methods A trauma-pain model was established in Wistar rats by combining right limb amputation with 50 ℃ tail-flick test.The contents of MDA and the activity of the total superoxide dismutase(SOD)in the brain tissue of trauma-pain rats were detected by thiobarbituric acid method and the xanthine oxidase method.A physical dependence model in mice was reproduced by subcutaneous injection of pethidine and the withdrawal syndromes were induced by intraperitoneal injection of naloxone.Results The contents of MDA enhanced,but the activity of SOD decreased greatly in brain tissues postinjury in rats.Melatonin(30,60,120 mg·kg-1)i.p.reduced the contents of MDA and enhanced the activity of SOD dose-dependently.Melatonin alone showed no withdrawal syndrome when it was given until the dosage of 840 mg·kg-1.Moreover,Melatonin(5,15,20 mg·kg-1)obviously inhibited the withdrawal jumpings of mice in pethidine-treated groups dose-dependently(P<0.01),the jumping rates of mice were decreased 61.4%,72.8%,84.8%,respectively.Conclusions The present results indicated that melatonin has good antioxidative effects on the trauma rats.In addiction,melatonin might inhibit withdrawal syndromes in pethidine-dependent mice.     

Study on mechanisms of hypertension in rat adult offspring following prenatal exposure to immuno-inflammatory stimulants

Objective Essential hypertension(EH)is one of the most common cardiovascular disease and the main causes of human fatility.Recently significant progress has been made in our lab,it was found that exterior stimulation during pregnancy may play a key role in chronicle adult disease.However,what factors affect the growth of fetus after those exterior stimulation and why has not been reported.Based on our previous finding,this study intends to investigate how immuno-inflammatory stimulation affect the development of embryo.Methods 1.Sprague-Dawley(SD)rats,dams in each group received i.p.injections of 0.79 mg·kg-1 LPS,8 mg·kg-1 zymosan or sterile saline respectively on their gestational days 8,10,and 12.2.The serums were collected in tail nick at 2 h after the last injection,and the amniotic fluid was mixed at 2,12,24,48 h after the last injection.TNF-α and IL-6 levels of serum and amniotic fluid were measured by RIA method.3.TNF-α and IL-6 mRNA levels were quantitated in amnion,placenta,amniotic fluid,Embryo and macrophage by real-time fluorescent quantitative-PCR.Results 1.The serum level of TNF-α and IL-6 in LPS group and zymosan group was higher than that in control group(P<0.01).It showed that there was immuno-imflammatory response after LPS or zymosan injection in rats.The mRNA levels of TNF-α and IL-6 was very higher in macrophage than in other organization.2.In embryo,the mRNA level of IL-6 was more than other organization,but the mRNA level of TNF-α was lower than other organization.However,the IL-6 mRNA level of LPS group and zymosan group was higher several dozens times than control group on 24 h and 48 h.Conclusions It suggested that IL-6 was important in the model that prenatalexposure to immuno-inflammatory stimulant results in increases of blood pressure and body weight in rats.