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Objective:To investigate the effect of minocycline on neuroinflammation of rats with post-traumatic stress disorder(PTSD).Methods:The rat model of PTSD was prepared by a single prolonged stress(SPS)method,and the rats were treated with minocycline(PTSD+Mino group)or normal saline(PTSD group)by gavage.The behavioral changes of rats were detected by light-dark box test.The expression of ionized calcium-binding adapter molecule 1(Iba-1)in hippocampus was detected by immunohistochemical staining.The contents of IL-1β and TNF-α in hippocampus were detected by ELISA,and the expression levels of IL-1β and TNF-α mRNA in hippocampus were detected by real-time RT-PCR(qRT-PCR).Results:After 3 days of SPS stimulation,the anxiety-like behavior of rats was obvious,the expression of Iba-1 in hippocampus was increased,and the contents of IL-1β and TNF-α in hippocampus were in-creased.Minocycline treatment significantly reduced anxiety-like behavior and decreased the expression of Iba-1 in the hippocampus of PTSD rats.Meanwhile,minocycline treatment also decreased the levels of IL-1β and TNF-α mRNA and protein in the hippocampus.Conclusion:Minocycline can improve the anxiety-like behavior of PTSD rats by inhibiting the activation of microglia.
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Objective:To study the effects of Shuxuening injection(SXN)on Wnt/β-catenin signaling pathway in hippocampus of rats with cerebral ischemia.Methods:SD rats were divided into three groups:Sham operation group(Sham),middle cerebral artery occlusion group(MCAO)and MCAO+SXN treatment group(MCAO+SXN).The model of cerebral ischemia in rats was prepared by MCAO.The rats with cerebral ischemia were treated with SXN by caudal vein injection.Zea-Longa scoring criteria and balance beam test were employed to evaluated neurological function of rats.HE staining were used to observe the changes of inflammatory cells infiltration the hippocampal CAI region.The expression of β-catenin in hippocampal CA1 region was observed by immunofluorescence staining.The mRNA and pro-tein expressions of caspase-3,cyclooxygenase-2(COX-2)and endothelial nitric oxide synthetase(eNOS)in hippocam-pus were detected by real time RT-PCR and Western Blot,respectively.Results:SXN can SXN can improve the neuro-logical dysfunction of cerebral ischemia rats.The inflammatory cells infiltration in hippocampal CAI region was decreased,and the expression of β-catenin,caspase-3 and COX-2 was decreased,while the expression of eNOS was upregulated.Conclusion:SXN protects against cerebral ischemia by inhibiting Wnt/β-catenin signaling pathway against inflammation response,oxidative stress and apoptosis of nerve cells in rats.
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Objective To observe the effect of Shenghui Granule on EC and CA1 regions of scopolamine-induced dementia rats and explore its mechanism based on p38 MAPK signal pathway.Methods 40 SD rats were randomly divided into four groups(blank group,model group,Shenghui granule group,donepezil group),and were treated with scopolamine.Morris water maze and open field test were used to evaluate the cognition and anxiety behavior of rats.The nerve injury of EC and CA1 was observed by HE staining.The activity of neurons in EC and CA1 regions was observed by c-Fos immunofluorescence staining.Western blot was used to detect p38 MAPK pathway related proteins.Results The behavioral experiment found that Shenghui Granule could improve the cognitive impairment and anxiety-like behavior of AD model rats.The results of HE staining showed that Shenghui granules had protective effects on EC and CA1 regions.The results of c-Fos immunofluorescence staining showed that Shenghui granules could increase the activity of neurons in EC and CA1 regions.Western blot results showed that Shenghui Granule could down-regulate the expression of Bax,reduce the levels of phosphorylated p38 and Tau,and increase the expression of Bcl-2.Conclusion Shenghui granule has protective effect on EC and CA1 regions of AD model rats,and may play a therapeutic role through p38 MAPK signal pathway.
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Objective To analyze the influence of cyclic RNA homologous domain interacting protein kinase 3(circ_HIPK3)on function and morphology of myloid β-protein(Aβ)induced hippocampal neurons by targeting miR-381-3p/zinc finger protein 217(ZNF217)axis.Methods Hippocampal neurons of neonatal rats were prepared and divided into the control group,the Aβ group,the si NC1 group,the si HIPK3 group,the si HIPK3+inhibitor NC group,the si HIPK3+miR-381-3p inhibitor group,the si HIPK3+miR-381-3p inhibitor+si NC2 group and the si HIPK3+miR-381-3p inhibitor+si ZNF217 group.Except the control group,all the other groups were modeled by 40 μmol/L Aβ1～42.qRT-PCR was used to determine the circ of hippocampal neurons circ_HIPK3,miR-381-3p and ZNF217 mRNA levels.Cell morphology was observed by transmission electron microscope,and the survival rate of hippocampal neurons was measured by CCK-8 method.Hochesst 33342 method was used to measure apoptosis of hippocampal neurons.The intracellular Ca2+ fluorescence intensity of hippocampal neurons was detected by flow cytometry.The expression levels of P-Tau,B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),Caspase-3 and ZNF217 proteins in hippocampal neurons were measured by Western blot assay.Double luciferase reporter genes were used to analyze the targeting relationship between miR-381-3p and circ_HIPK3,ZNF217.Results In the control group,the structure of hippocampal neurons was normal,the morphology of nucleus was normal,and there were no pathological changes in mitochondria and endoplasmic reticulum.In the Aβ group,hippocampal neurons showed degenerative changes,abnormal nuclear morphology,membrane invagination,a large number of mitochondria swelling and a large number of lipid droplets vacuoles in cytoplasm.Compared with the Aβ group,the hippocampal neuronal structure was partially restored in the si HIPK3 group.Compared with the si HIPK3 group,the hippocampal neuronal structure was severely damaged in the si HIPK3+miR-381-3p inhibitor group.Compared with the si HIPK3+miR-381-3p inhibitor group,the damage of hippocampal neurons in the si HIPK3+miR-381-3p inhibitor+si ZNF217 group was reduced.Compared with the control group,the circ_HIPK3,ZNF217 mRNA and ZNF217 protein levels,apoptosis rate,Ca2+ fluorescence intensity,P-Tau,Bax,Caspase-3 protein expression of hippocampal neurons were increased in the Aβ group,and the miR-381-3p level,survival rate and Bcl-2 protein expression decreased(P＜0.05).Compared with the Aβ group,the circ_HIPK3,ZNF217 mRNA and ZNF217 protein levels,apoptosis rate,Ca2+ fluorescence intensity,P-Tau,Bax and Caspase-3 protein expression of hippocampal neurons were decreased in the si HIPK3 group,and miR-381-3p level,survival rate and Bcl-2 protein expression increased(P＜0.05).Compared with the si HIPK3 group,the circ_HIPK3,ZNF217 mRNA and ZNF217 protein levels,apoptosis rate,Ca2+ fluorescence intensity,P-Tau,Bax and Caspase-3 protein expression of hippocampal neurons in the si HIPK3+miR-381-3p inhibitor group were increased,and the miR-381-3p level,survival rate and Bcl-2 protein expression decreased(P＜0.05).Compared with the si HIPK3+miR-381-3p inhibitor group,the ZNF217 mRNA and ZNF217 protein levels,apoptosis rate,Ca2+ fluorescence intensity,P-Tau,Bax and Caspase-3 protein expression of hippocampal neurons in the si HIPK3+miR-381-3p inhibitor+si ZNF217 group were decreased,and the survival rate and Bcl-2 protein expression increased(P＜0.05).miR-381-3p targeted and combined with HIPK3 and ZNF217.Conclusion circ_HIPK3 silencing may ameliorate Aβ-induced damage of hippocampal neuronal structure and function by regulating miR-381-3p/ZNF217 axis.
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Objective:Utilizing functional magnetic resonance imaging (fMRI) to investigate changes in brain structure and function in patients with alcohol dependence (AD) complicated by major depressive disorder (MDD), and assessing the clinical significance of fMRI in diagnosing alcohol dependence complicated by MDD.Methods:From August 2019 to October 2022, 90 patients with AD complicated by MDD and 90 healthy subjects who concurrently received physical examination in our hospital were included in the study. All participants underwent magnetic resonance imaging (MRI) and fMRI to observe the brain tissue structure of patients with AD complicated by MDD and to assess differences in N-acetylaspartic acid/creatine (NAA/Cr) and factional anisotropy (FA) values across different brain tissue regions.Results:The widths of the left and right ventricular temporal angles in the AD complicated by MDD group [(2.67 ± 0.24) mm, (2.63 ± 0.25) mm] were significantly higher than those observed in the healthy control group [(2.29 ± 0.21) mm, (2.31 ± 0.23) mm, t = 22.48, 20.64, both P < 0.001]. Additionally, the volumes of the left and right hippocampus and nucleus accumbens in the AD complicated by MDD group [(2 673.46 ± 155.74) mm 3, (2 692.29 ± 154.61) mm 3, (682.04 ± 65.37) mm 3, (729.65 ± 68.49) mm 3] were significantly lower compared with those in the healthy control group [(2 826.53 ± 158.95) mm 3, (2 849.17 ± 157.23) mm 3, (766.28 ± 69.51) mm 3, and (805.43 ± 71.36) mm 3, t = -9.53, -8.44, -15.62, -13.92, all P < 0.001]. Moreover, the NAA/Cr values in the left and right frontal lobes, temporal lobes, hippocampi, and nucleus accumbens in the AD complicated by MDD group were significantly lower than those in the healthy control group ( t = -11.36, -7.19, -9.96, -7.84, -14.59, -8.25, -7.64, -6.84, all P < 0.001). Similarly, the FA values of the left and right frontal lobes, temporal lobes, hippocampi, and right nucleus accumbens in the AD complicated by MDD group were significantly lower compared with those in the healthy control group ( t = -9.48, -11.74, -9.22, -10.36, -16.85, -14.67, -5.28, all P < 0.001). Conclusion:Patients with AD accompanied by MDD exhibit alterations in brain tissue structure, neuronal metabolic function, and the integrity of white matter nerve fibers. fMRI is effective in identifying changes in brain neuron metabolism and the integrity of white matter nerve fibers, making it invaluable for the diagnosis and assessment of AD accompanied by MDD.
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Objective:To investigate the effect and the neural mechanisms of Apelin-13 on the behavior changes of posttraumatic stress disorder (PTSD) model mice.Methods:Totally 32 SPF grade male C57BL/6J mice aged 6 weeks were divided into 4 groups randomly ( n=8 in each group): control group, model group, normal saline group and Apelin-13 group.The mice model of PTSD was established by single-prolonged stress (SPS) method. The mice in normal saline group and Apelin-13 group were respectively given lateral ventricular microinjection of 0.9% sodium chloride solution (2 μL) and Apelin-13 (1.5 μg/μL, 2 μL)after PTSD modeling. The behaviors of mice were evaluated by open field test, elevated plus maze test and Morris water maze test.The morphological structure and numerical changes of hippocampal neurons were observed by hematoxylin and eosin (HE) staining.The expression of phosphoinositide 3-kinase(PI3K), phosphorylated-PI3K(p-PI3K), protein kinase B(Akt), phosphorylated-Akt (p-Akt), forkhead box O3a (FoxO3a), phosphorylated-FoxO3a(p-FoxO3a), autophagy-related proteins including microtubule-associated protein 1 light chain 3(LC3) and sequestosome 1(p62) were detected by Western blot. SPSS 26.0 software was used for data analysis.The escape latency data of repeated learning training in Morris water maze was conducted by repetitive measurement ANOVA.The comparison of other data among multiple groups was conducted by one-way ANOVA and further pairwise comparisons were conducted by LSD test and Tamhane test. Result:(1) Open field test results showed statistically significant differences in the central area activity distance and residence time in central area among mice in the four groups ( F=15.37, 9.63, both P<0.05). The central area activity distance ((0.06±0.03) m) and residence time ((2.48±1.02) s) of the mice in model group were lower than those of the control group ((0.19±0.05) m, (15.00±8.91) s)(both P<0.05). And the central area activity distance((0.12±0.04)m)and the residence time((13.56±7.64)s)were higher than those of model group((0.06±0.03)m, (2.48±1.02)s)and normal saline group((0.06±0.02)m, (2.82±1.52)s)(all P<0.05). Elevated plus maze test results showed statistically significant differences in the numbers and time entering open arms among the four groups ( F=10.74, 19.12, both P<0.05). The numbers((4.50±2.51) times) and the time ((26.95±17.48) s) entering the open arm of mice in model group were both lower than those of the control group ((13.75±4.71) times, (103.75±42.43)s) and Apelin-13 group ((10.00±5.18) times, (55.98±19.49) s) (all P<0.05). Morris water maze test results showed that in the 4-day learning and training phase, the time and group interaction of escape latency was not significant among the four groups ( F=1.15, P=0.34), but time main effect and group main effect were significant ( F=131.65, 16.98, both P<0.05). On the 2nd to 4th day, mice in model group showed significantly increased escape latency than mice in control group and Apelin-13 group(both P<0.05). And the numbers crossing original platform and the time in the target quadrant of Apelin-13 group were both higher than those of model group and normal saline group (all P<0.05). (2) HE staining results showed that neurons in the hippocampal CA1 and CA3 area of mice in model group and normal saline group were swollen and arranged loosely.The hippocampal neurons in control group and Apelin-13 group were arranged neatly and densely. (3) Western blot results showed statistically significant differences in the protein expression of p-PI3K, p-Akt, p-FoxO3a, p62 and the ratio of LC3Ⅱ/LC3Ⅰ among the four groups ( F=21.37, 37.35, 20.71, 13.26, 37.65, all P<0.05). The protein expression of p-PI3K, p-Akt, p-FoxO3a and p62 in Apelin-13 group((0.92±0.07), (0.90±0.09), (0.89±0.13), (1.03±0.08)) were higher than those in model group((0.59±0.04), (0.50±0.07), (0.49±0.11), (0.68±0.04)) and normal saline group((0.61±0.06), (0.50±0.08), (0.53±0.11), (0.70±0.05))(all P<0.05), and the ratio of LC3Ⅱ/LC3Ⅰ in Apelin-13 group(0.60±0.06) was lower than those in model group(0.92±0.10) and normal saline group(0.99±0.05) (both P<0.05). Conclusion:Apelin-13 can alleviate the anxiety-like behavior and impaired spatial learning and memory in PTSD model mice. The mechanism may be related to the up-regulation of PI3K/Akt/FoxO3a autophagy pathway.
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Objective:To investigate the expression of hippocampal synapse-related proteins including synaptophysin (SYN), postsynaptic density protein 95 (PSD95) and growth-associated protein 43 (GAP43) in rats with epilepsy accompanied by depression.Methods:The 3-month-old female clean grade SD rats were selected for the experiment.Lithium chloride pilocarpine was used to establish an epileptic rat model. Rats with successful epilepsy models were divided into epileptic depressive group (EWD group)and epileptic group with 10 in each group based on whether they were accompanied by depression. Furthermore, ten rats with matched body mass were taken as the depressive group and 10 were taken as control group. As for the depressive group rats, chronic unpredictable mild stress stimulation combined with orphanage was adopted to establish a model of depression.The depressive behaviors of rats were evaluated by body mass, sucrose preference test and open field test. Immunohistochemical staining and Western blot were used to detect the expression of SYN, PSD95 and GAP43 proteins in rat hippocampal tissue. SPSS 17.0 software was used for data statistical analysis, repeated measurement ANOVA was used for behavioral results, one-way ANOVA was used for inter group comparison of protein expression data, and LSD test was used for further pairwise comparison.Results:As for the body mass, there was significant interaction effect between the time and group among the 4 groups ( F=7.33, P<0.01). On the 8th day and the 29th day, the body weight of rats in the EWD group and the depressive group were lower than those in the epilepsy group (all P<0.05). The body weight of rats in the EWD group on the 29th day was lower than that on the first day ( P<0.05). As for the sucrose preference rates, there was significant interaction effect between the time and group among the 4 groups( F=2.67, P<0.05). The sucrose preference rate of EWD group on the15th and 29th day were lower than that on the first day (both P<0.05). The results of the open field test showed that the interaction effects of the number of vertical standing times( F=2.74) and the number of horizontal movement lattices ( F=1.76) both were not significant (both P>0.05), but both the time effect and group effect were significant (vertical standing times: Ftime=4.35, P<0.05, Fgroup=25.64, P<0.01; horizontal movement lattices: Ftime=12.75, P<0.01, Fgroup=21.37, P<0.01). The immunohistochemical results showed that there was a statistically significant difference in the number of positive cells expressing synaptic proteins SYN, PSD95 and GAP43 among the four groups of rats ( F=93.85, 58.66, 98.84, all P<0.05). The numbers of positive cells of SYN (11.73±4.30), PSD95 (24.47±7.58) and GAP43 (9.40±3.50) in the epilepsy group were lower than those in the control group ((51.00±15.39), (55.60±13.17) and (29.53±4.05)) (all P<0.05). The numbers of positive cells of SYN (5.80±3.53), PSD95 (12.87±4.03) and GAP43 (5.33±3.50) in the EWD group were lower than those in the depressive group ((11.33±3.22), (48.13±12.69) and (15.47±5.21) )(all P<0.05). Western blot results showed that there were statistically significant differences in the expression of synaptic proteins SYN, PSD95 and GAP43 among the four groups of rats ( F=13.19, 9.38, 16.80, all P<0.05). The expression levels of SYN, PSD95 and GAP43 in the epilepsy group were lower than those in the control group (all P<0.05). The expression levels of SYN, PSD95 and GAP43 in the EWD group were lower than those in the epilepsy group and the depressive group (all P<0.05). Conclusion:The low expression of SYN, PSD95 and GAP43 proteins in the hippocampus of rats with epilepsy accompanied by depression may be related to their pathogenesis.
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Objective:To investigate the effect and mechanism of different doses of luteolin on memory function and apoptosis-related proteins of aging rats induced by D-galactose.Methods:Forty-eight SPF-grade male Wistar rats aged 6-8 weeks were randomly divided into control group, model group, luteolin low-dose group (25 mg/kg), medium-dose group (50 mg/kg), high-dose group (100 mg/kg), and vitamin C group (100 mg/kg), with 8 rats in each group. D-galactose (1 000 mg/kg) was subcutaneously injected to establish the aging rat model, while luteolin was used for preventive treatment. The Morris water maze test was used to evaluate the learning and memory abilities of the rats.Transmission electron microscopy was used to detect the morphology of hippocampal neurons in rats.Spectrophotometry was used to detect the levels of interleukin-6 (IL-6), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), superoxide dismutase (SOD), malondialdehyde (MDA), and the total antioxidant capacity (T-AOC). RT-PCR was used to detect miR-34a mRNA expression.Western blot technique was used to detect the expression levels of silent regulator protein 1 (SIRT1), B-cell lymphoma-2 (Bcl-2), cleaved caspase-3, p53, and p21. Statistical analysis was performed using SPSS 22.0, and one-way ANOVA was used for multi-group comparison, followed by LSD- t test for further pairwise comparisons. Results:(1) The differences in escape latency among the 6 groups of rats were statistically significant ( F=120.93, P<0.001). The latency of first finding the platform location of the model group rats ((54.61±3.60) s) was higher than that of the control group ((10.54±4.27) s) ( P<0.05). The latency of first finding the platform location of rats in the low, medium and high dosage groups of luteolin ((45.50±3.81)s, (37.46±2.94) s, (32.32±3.14) s) was lower than that of the model group ((54.61±3.60) s) (all P<0.05). (2) The differences of SOD, MDA, T-AOC, TNF-α, IL-1β, and IL-6 levels in the cerebral cortex of the 6 groups of rats were all statistically significant ( F=281.636, 75.119, 208.228, 38.999, 28.428, 52.767, all P<0.001). Compared with the control group, the model group showed abnormal levels of inflammatory factors and antioxidant indexes. In the medium and high dosage groups of luteolin, the SOD and T-AOC contents in the cerebral cortex of rats were higher than those in the model group (all P<0.05), while the levels of MDA, TNF-α, IL-1β, and IL-6 were lower than those in the model group (all P<0.05). (3) The differences in relative expression levels of miR-34a mRNA among the 6 groups of rats were statistically significant ( F=81.439, P<0.001). The expression levels of miR-34a mRNA in the hippocampal tissues of rats in the luteolin treatment group were lower than those in the model group ( P<0.05). (4) The differences in protein expression levels of SIRT1, p53, and p21 in the hippocampal tissues of the 6 groups of rats were statistically significant ( F=159.946, 38.342, 123.608, all P<0.001). The expression levels of p53 and p21 in the medium and high dosage groups of luteolin were lower than those in the model group (all P<0.05), while the expression level of SIRT1 protein was higher than that in the model group ( P<0.05). (5) The differences in protein expression levels of Bcl-2 and cleaved caspase-3 in the hippocampal tissues of the 6 groups of rats were statistically significant ( F=112.659, 43.296, both P<0.05). The expression levels of Bcl-2 in the low, medium, and high dosage groups of luteolin ((0.24±0.04), (0.40±0.03), (0.48±0.05) pg/μg) were higher than those in the model group ((0.09±0.06) μg) ( P<0.05), while the expression levels of cleaved caspase-3 in the low, medium, and high dosage groups of luteolin ((0.62±0.04), (0.61±0.09), (0.51±0.10) μg) were lower than those in the model group ((0.75±0.05) μg) ( P<0.05). Conclusions:Luteolin can alleviate cellular oxidative damage through downregulating the miR-34a SIRT1/p53 signaling pathway and reducing cell apoptosis.
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OBJECTIVE To investigate the effect and mechanism of acute exposure to sodium cyanide(NaCN)on brain nerve damage induced by closed hypoxia in mice.METHODS ① Mice were randomly divided into hypoxia+NaCN 0(hypoxia control group),2.56,3.8,and 5.1 mg·kg-1 groups.After ip adminis-tration of different concentrations of NaCN,the mice were immediately placed into a closed hypoxic tank and the hypoxia survival time was observed.②Mice were divided into normal control,NaCN 3.8 mg·kg-1,hypoxia(30 and 60 min)and NaCN 3.8 mg·kg-1+hypoxia(30 and 60 min)groups.After grouping,the pH,oxygen saturation(sO2),oxygen tension(pO2)and carbon dioxide partial pressure(pCO2)of arterial blood of mice were detected using an arterial blood gas analyzer.The cortical cerebral blood flow of mice was detected using a laser speckle imager.The dry and wet brain tissue were weighed separately,and the brain moisture content was calculated.The kit was used to detect the activity of total superoxide dismutase(T-SOD)and the content of malondialdehyde(MDA)in the hippocampus.TUNEL staining was used to detect the apoptosis rate of cells in the hippocampus.HE staining was used to detect path-ological changes in the hippocampus.RESULTS ①Compared with the hypoxic control group,the sur-vival time of mice in the hypoxic+NaCN groups was significantly prolonged(P<0.01).②Compared with the normal control group,the hypoxia 30 min group showed upregulation of arterial blood p CO2(P<0.05),downregulation of p O2(P<0.05).The hypoxia 60 min group showed upregulation of arterial blood p CO2(P<0.05)and downregulation of cortical cerebral blood flow(P<0.05).In the NaCN 3.8 mg·kg-1 group,arterial blood p O2 and s O2 were significantly downregulated(P<0.05),so was cortical cerebral blood flow(P<0.01),but MDA content and T-SOD activity were significantly upregulated(P<0.01),and the brain moisture content was increased(P<0.01).Compared with the hypoxia 30 min group,s O2 and p O2 of arterial blood in the NaCN+hypoxia 30 min group were significantly upregulated(P<0.05),while p CO2 was significantly downregulated(P<0.05).Compared with the hypoxia group at corresponding time points,the NaCN+hypoxia 30 or 60 min groups showed significant downregulation of cerebral blood flow(P<0.01),significant upregulation of MDA content and T-SOD activity(P<0.01),and signifi-cant upregulation of brain moisture content(P<0.01).HE staining results showed that the NaCN 3.8 mg·kg-1 group and the NaCN+hypoxia group(30 or 60 min)showed significant cell swelling and vacuolization in cells in the hippocampal tissue,a decrease in the number of neurons,nuclear pyknosis and deep staining.TUNEL fluorescence results showed that the NaCN 3.8 mg·kg-1 group significantly increased the apop-tosis rate of the mouse hippocampus compared with the normal control group(P<0.05).The NaCN+ hypoxia 30 and 60 min groups significantly increased the apoptosis rate of the mouse hippocampus compared with the hypoxia group at corresponding time points(P<0.05).CONCLUSION NaCN can exacerbate hypoxia induced decrease in cerebral blood flow,oxidative stress in brain tissue,and neuro-nal apoptosis in mice,thereby reducing oxygen consumption in closed hypoxic tanks and prolonging their survival time.The mechanism is related to reduced utility of cell oxygen,delaying CO2 accumulation and increasing free oxygen in vivo.
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Objective To observe the effects of electroacupuncture(EA)on glutamate(Glu),metabotropic glutamate receptor 2/3(mGluR2/3)and apoptosis related proteins expression in hippocampus in rats with acute myocardial ischemia(AMI);To explore the mechanism of EA against AMI.Methods Totally 50 SD rats were randomly divided into sham-operation group,model group,EA group and inhibitor group,with 10 rats in each group.Except for the sham-operation group,the rats were treated with ligation at the left anterior descending coronary artery to establish AMI model.The rats in the EA group was treated with EA at"Shenmen"and"Tongli",30 minutes each time,once a day for 3 consecutive days.The rats in the inhibitor group were treated with injection of LY341459 via the lateral ventricle 30 min after modeling.HE staining was used to observe myocardial tissue morphology,and ELISA was used to detect Caspase-3 activity in myocardial tissue and Glu content in hippocampal tissue,immunofluorescence staining was used to detect mGluR2/3 expression in hippocampal tissue,TUNEL staining was used to detect apoptosis in hippocampal tissue cells,Western blot was used to detect the expressions of PI3K,Akt,and Caspase-3 protein in hippocampal tissue.Results Compared with the sham-operation group,the myocardial cells of the model group rats showed sparse and swelling with severe infiltration of inflammatory cells;the activity of Caspase-3 in myocardial tissue significantly increased,and the Glu content,positive expression of mGluR2/3,number of apoptotic cells in hippocampal tissue significantly increased(P<0.01),and the expressions of PI3K and Akt proteins in hippocampal tissue were significantly decreased,while the expression of Caspase-3 protein significantly increased(P<0.01).Compared with the model group,myocardial cell edema and inflammatory cell infiltration were reduced in the EA group and inhibitor group,the activity of Caspase-3 in myocardial tissue was significantly decreased,the Glu content,positive expression of mGluR2/3,and number of apoptotic cells in hippocampal tissue were significantly reduced(P<0.01),the expressions of PI3K and Akt proteins in hippocampal tissue significantly increased,while the expression of Caspase-3 protein significantly decreased(P<0.01).Conclusion EA can improve myocardial injury in AMI rats,and its mechanism may be related to activation of PI3K/Akt signaling pathway,inhibition of hippocampal mGluR2/3 overexpression,reduction of Glu accumulation,inhibition of apoptosis of hippocampal neurons and reduction of neurotoxicity.
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Objective To explore the effects and mechanism of Baishile Capsules regulating SHH/Gli1 signaling pathway on hippocampal neurogenesis of depression model rats.Methods Totally 32 SD rats were randomly divided into control group,model group,fluoxetine(5.4 mg/kg)group and Baishile Capsules(2.88 g/kg)group,with 8 rats in each group.A depression rat model was established using chronic unpredictable mild stress and single cage feeding method.The model was established and administered simultaneously for 21 consecutive days.Depression-like behavior in rats were evaluated by sucrose preference experiment and open field experiment,ELISA was used to detect brain derived neurotrophic factor(BDNF)contents in rat serum and hippocampal tissue,the number of BrdU,BrdU/DCX,BrdU/NeuN positive cells in dentate gyrus of the hippocampus was observed by immunofluorescence,immunofluorescence and Western blot were used to detect the fluorescence intensity and protein expression of SHH,Gli1,Smo,Ptch in hippocampal tissue.Results Compared with the control group,the degree of sucrose preference significantly decreased in the model group(P<0.01),the number of horizontal and vertical movements significantly decreased(P<0.01),the contents of BDNF in serum and hippocampal tissue significantly decreased(P<0.05),the number of BrdU,BrdU/DCX,BrdU/NeuN positive cells in dentate gyrus of the hippocampus significantly decreased(P<0.01),and the fluorescence intensity and protein expression of SHH,Gli1,Smo,Ptch in hippocampal tissue significantly decreased(P<0.01,P<0.05).Compared with the model group,the degree of sucrose preference and the number of horizontal and vertical movements in fluoxetine group and Baishile Capsule group increased significantly(P<0.05,P<0.01),the contents of BDNF in serum and hippocampal tissue significantly increased(P<0.05,P<0.01),and the number of BrdU,BrdU/DCX,BrdU/NeuN positive cells in dentate gyrus of the hippocampus significantly increased(P<0.01,P<0.05),the fluorescence intensity and protein expressions of SHH,Gli1,Smo,Ptch in hippocampal tissue significantly increased(P<0.01,P<0.05).Conclusion Baishile Capsule can promote the hippocampus neurogenesis in depression model rats by regulating SHH/Gli1 signaling pathway,and play an antidepressant role.
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Objective:To analyze the treatment efficacy, safety and dose parameters of optimized hippocampus-avoidance prophylactic cranial irradiation (HA-PCI) in limited-stage small cell lung cancer (LS-SCLC) and explore the corresponding dosimetric parameters under the condition of narrowing the hippocampus avoidance region as hippocampus region plus 2 mm in three dimensions.Methods:Clinical data of patients with LS-SCLC receiving HA-PCI (hippocampus avoidance region defined as hippocampus region plus 2 mm in three dimensions) in Cancer Hospital Chinese Academy of Medical Sciences from August 2014 to June 2020 were retrospectively analyzed. Dose parameters of HA-PCI and adverse events were analyzed using descriptive statistics analysis. Changes of neurocognitive function, such as mini-mental state examination (MMSE) and Hopkins verbal learning test-revised (HVLT-R) scores, were evaluated by analysis of variance and Kruskal-Wallis H test. Overall survival (OS), progression-free survival (PFS) and intracranial PFS (iPFS) were calculated using Kaplan-Meier method. The cumulative incidence of local-regional recurrence (LRR), extracranial distant metastases (EDM), and locoregional recurrence (LR) were investigated under competing risk analysis. Results:A total of 112 patients were included, the median follow-up time was 50 months (95% CI: 45.61-54.38). The median volume of hippocampus was 4.85 ml (range: 2.65-8.34 ml), with the average dose ≤9 Gy in 106 patients (94.6%), ≤8 Gy in 92 patients (82.1%). The median volume of hippocampus avoidance area was 15.00 ml (range: 8.61-28.06 ml), with the average dose ≤12 Gy in 109 patients (97.3%), ≤10 Gy in 101 patients (90.2%). The 2-year cumulative LRR, EDM, LR rates were 16.9%, 23.2% and 28.5%, respectively. The 5-year cumulative LRR, EDM, LR rates were 23.2%, 26.9% and 33.3%, respectively. The 2-year iPFS, PFS and OS rates were 66.1% (95% CI: 57.9%-75.4%), 53.6% (95% CI: 45.1%-63.7%) and 80.4% (95% CI: 73.3%-88.1%), respectively. The most common grade I-Ⅱ adverse events were nausea (33.9%) and dizziness (31.3%), and only 1 patient developed grade Ⅲ nausea and dizziness. MMSE ( n=57) and HVLT-R tests ( n=56) showed no significant decline. Conclusions:Optimized HA-PCI can achieve similar dose limitation with favorable efficacy and light toxicity. No significant decline is observed in short-term neurocognitive function in evaluable patients.
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The hippocampus and its circuits play crucial roles in human learning, memory, and emotional regulation. Whether it is vascular cognitive impairment (VCI) or Alzheimer's disease (AD), damage to the hippocampus is a prominent pathological feature. This review summarizes the recent advance in multimodal magnetic resonance imaging in anatomy, blood supply, structure and function of the hippocampus and the circuits related to VCI and AD in recent years, aiming to provide help in early recognizing and differentially diagnosing VCI and AD.
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ObjectiveTo investigate the intervention effect of heat shock protein 60 (HSP60) on learning and memory impairment induced by combined exposure to lead and hypertension in mice, and the relative mechanism of triggering receptor expressed on myeloid cells 2 (TREM2). Methods Specific pathogen-free C57BL/6J male mice were randomly divided into control group, hypertension group, lead-exposed group and lead-exposed + hypertension group, or into control group, heat shock protein 60 (HSP60) control group, lead-exposed + hypertension group and HSP60 intervention group, with 10 mice in each group. Mice of hypertension group and lead-exposed + hypertension group were intraperitoneally injected with angiotensin Ⅱ at a dose of 0.5 mg/(kg·d) for seven consecutive days to induce hypertension model. Mice of the lead-exposed group, lead-exposed + hypertension group, and HSP60 intervention group were given lead acetate drinking water with a mass concentration of 250.0 mg/L, while mice in the control group, hypertension group, and HSP60 control group were given purified water for 12 weeks. Mice of the HSP60 control group and HSP60 intervention group were intraperitoneally injected with a solution of HSP60 at a dose of 4 mg/kg body weight, every other day for a total of three times at the 12th week. The learning and memory ability of mice was detected using the Morris water maze test. The enzyme-linked immunosorbent assay was used to detect the levels of interleukin (IL)-1β, IL-6 and tumor necrosis factor-α (TNF-α) in the hippocampal tissues of the mice. The relative expression of ionized calcium binding adaptor molecule-1 (IBA1) and TREM2 protein in the hippocampus of mice was detected using Western blot. Results i) The number of platform crossings of the mice in the hypertension group and the lead-exposed group was lower than that in the control group (both P<0.05). The escape latency of the mice on the third day was longer and the number of platform crossings was lower in the lead-exposed + hypertension group compared with the control group, hypertension group and lead-exposed group (all P<0.05). The levels of IL-1β, IL-6, and TNF-α in the hippocampus of the other three groups increased compared with the control group (all P<0.05). The relative expression of IBA1 protein in the hippocampus of lead-exposed group and lead-exposed + hypertension group increased (all P<0.05), while the relative protein expression of TREM2 decreased compared with the control group (all P<0.05). The levels of IL-1β, IL-6, TNF-α, and the relative protein expression of IBA1 protein in the hippocampus of the lead-exposed+hypertension group were higher (all P<0.05), and relative expression of TREM2 protein was lower (P<0.05) than those in the hypertension group. The level of TNF-α and the relative expression of IBA1 protein in the hippocampus of lead-exposed+hypertension group were higher than those in lead-exposed group (all P<0.05). ii) The escape latency of mice in the lead-exposed + hypertension group was longer than that in the control group (P<0.05), and the number of platform crossings was fewer than that in the control group (P<0.05). The escape latency of mice in the HSP60 intervention group was shortened (P<0.05), the number of platform crossings increased (P<0.05), and the levels of IL-1β, IL-6, TNF-α and relative expression of IBA1 protein decreased in the hippocampus (all P<0.05), while the relative expression of TREM2 protein increased (P<0.05) compared with the lead-exposed+hypertension group. Conclusion Combined exposure of lead and hypertension has a synergistic effect on learning and memory impairment in mice. The mechanism may be related to the inhibition of TREM2 expression by lead in the hippocampus of hypertensive mice and aggravating the neuroinflammatory response. Intervention with TREM2 receptor agonist HSP60 can alleviate learning and memory impairment in mice exposed to lead and hypertension by up-regulating TREM2 expression in the hippocampus.
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Depression is a prevalent mental illness worldwide, its multifaceted pathogenesis is still in the exploratory stage. MicroRNA (miRNA), as a crucial epigenetic regulator, plays an important role in depression. miR-124 is one of the most abundant miRNAs in the central nervous system including neurons and microglia, and involved in various biological events like neuron development and differentiation, synaptic and axonal growth, neural plasticity, inflammation and autophagy. Recent studies have reported abnormal expression of miR-124 in both depression patients and animal models. Most of the studies showed that miR-124 is upregulated in the hippocampus or prefrontal cortex in stress-induced rodent depression animal models such as CUMS, CSDS, CORT, CRS and LH but some evidence for divergence. Upregulation of miR-124 expression may be involved in depression-like behavior via CREB/BDNF/TrkB pathway, GR pathway, SIRT1 pathway, apoptosis and autophagy pathways by directly targeting these genes including Creb, Bdnf, Sirt1, Nr3c1, Ezh2 and Stat3. The downregulation of miR-124 expression in neurons is mainly involved in the neurogenesis and neuroplasticity impairments in depression by targeting the Notch signaling pathway and DDIT4/TSC1/2/mTORC1 pathway. The downregulation of miR-124 expression also was found in the activated microglia in the stress-induced models, and resulted in neuroinflammation. In summary, the abnormal expression of miR-124 in the brain of depression-related models and its related mechanisms are complex and even contradictory, and still need further research. This review provides a summary of the research progress of miR-124 in depression.
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Objective Based on the endoplasmic reticulum stress(ERS)inositol-requiring enzyme-1α(IRE1α)/apoptosis signal-regulated kinase 1(ASK1)/c-Jun N-terminal kinase(JNK)pathway to investigate the intervention effect of Ziziphi Spinosae Semen(ZSS)-Albiziae Flos(AF)on the depression model of rats,which were prepared by solitary rearing combined with chronic unpredictable mild stress(CUMS).Methods A total of 144 rats were randomly divided into normal group,model group,high-,medium-and low-dose groups of ZSS-AF,and Venlafaxine group.In addition to the normal group,the rats in other groups were isolated for 21 days combined with CUMS to prepare the depression model.The behavioral changes of rats were observed by open field test and Morris water maze test.The ultrastructural changes of hippocampal neurons were observed by transmission electron microscope.TUNEL staining was used to observe the apoptosis of nerve cells in hippocampus.The protein expression levels of IRE1α,phosphorylated(P)-IRE1α,ASK1,P-ASK1,JNK,P-JNK,B cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax)and cysteme aspartate specific protease-3(Caspase-3)in hippocampus were detected by Western Blot.The mRNA expression levels of IRE1α,ASK1,JNK,Bax,Bcl-2 and Caspase-3 in hippocampus were detected by Real-Time PCR.Results Compared with the normal group,the scores of horizontal movement and vertical movement in the open field test of rats in the model group were decreased(P＜0.01).In the water maze test,the escape latency was increased and the number of crossing the original platform was decreased(P＜0.01).The number of hippocampal apoptosis was increased(P＜0.01).The protein expression levels of P-IRE1 α/IRE1 α,P-ASK1/ASK1,P-JNK/JNK,Bax,Caspase-3 and mRNA expressions of IRE1α,ASK1,JNK,Bax,Caspase-3 in hippocampus were increased,while the protein and mRNA expression levels of Bcl-2 were decreased(P＜0.01).Compared with model group,the scores of horizontal movement and vertical movement in the open field test of rats in each dose ZSS-AF groups and Venlafaxine group were increased(P＜0.01).In the water maze test,the escape latency was decreased and the times of crossing the original platform was increased(P＜0.01).The number of hippocampal apoptosis was decreased(P＜0.01).The mRNA expression levels of P-IRE1α/IRE1α,P-ASK1/ASK1,P-JNK/JNK,Bax,Caspase-3 protein and IRE1α,ASK1,JNK,Bax,Caspase-3 in hippocampus were decreased,while the protein and mRNA expression levels of Bcl-2 were increased(P＜0.05,P＜0.01).The effect of medium-dose ZSS-AF group was better than that of high-and low-dose groups.Conclusion ZSS-AF may play an antidepressant role by regulating IRE1α/ASK1/JNK pathway of endoplasmic reticulum stress.
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@#Objective To analyze the expression profiles of long non-coding RNAs(lncRNA)in hippocampus of alcoholdependent mice induced by double-bottle selective drinking.Methods The alcohol-dependent mouse model was established by double-bottle selective drinking method,and the control group was set up(drinking water). Three male mice with alcohol preference more than 60% and alcohol consumption more than 10 g/(kg·24 h)in alcohol group and random three male mice in control group were selected,of which bilateral hippocampal brain tissues were isolated and stored in liquid nitrogen. LncRNA and mRNA of mouse hippocampal brain tissue RNA samples were sequenced by using Agilent-084388 microarray,and the differential expression of lncRNA in samples was detected by using ncRNA microarray. The biological processes and signaling pathways involved in differential expression of lncRNA were clustered and enriched by Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis. Pearson correlation analysis was used to predict the coding genes co-expressed by each differentially expressed lncRNA. Hypergeometric distribution test was used to calculate the significance of differential gene enrichment in each corresponding transcription factor item,and Cytoscape software was used to draw a visual network diagram.Results Compared with the control group,totally 855 lncRNAs(FC ≥ 2. 0,P < 0. 05)were differentially expressed in the hippocampus of mice in alcohol group,of which 337 lncRNAs were up-regulated significantly,with NONMMUT025786.2 and NONMMUT072246.2 being the most up-regulated,and 518 significant downward adjustments were observed,with the largest downward adjustments being NONMMUT113098.1 and NONMMUT076455.1. There were 361 mRNAs differentially expressed in the two groups(FC ≥ 2. 0,P < 0. 05)with 271 mRNAs up-regulated significantly and 90 significant downward adjustments,among which,the most obvious up-regulated were Upf3b and Zfp943,and Adamts 13 and Ift 27 showed the largest downward adjustments. The differential expression of lncRNAs was most obvious in the positive regulation of cell surface,GTPase activity and cell vesicle transport;The main signaling pathways involved were propanoate metabolism,taurine metabolism,extracellular matrix receptor interaction and AMPK signaling pathway. The most abundant transcription factors were FOXL1 and LHX3,with 25 and 21 corresponding co-expressed genes,respectively.Conclusion Through high-throughput gene expression profile microarray analysis,the possible key regulatory sites of lncRNAs and mRNAs were obtained,which provided experimental basis for research of the molecular mechanism of alcohol dependence in the hippocampus.
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Objective To investigate the effects of nuclear respiratory factor 1(NRF1)on mitochondrial and cog-nitive dysfunction in Alzheimer's disease(AD)model mice.Methods The 5 × FAD mice were utilized as a mod-el for Alzheimer's disease,and the sparsely labeled AAV virus overexpressing NRF1(AAV-NRF1)was adminis-tered via stereotaxic injection into the brain.The expression of NRF1 in hippocampus was determined by Western blot,the morphology of mitochondria in hippocampus was observed by transmission electron microscope,the den-dritic spines of sparsely labeled neurons in the CA1 region were visualized and quantified using confocal laser mi-croscopy,cognitive and memory functions of mice were evaluated using the Morris water maze test,while electro-physiological methods were employed to detect long-term potentiation(LTP)of synaptic efficacy.Results The ex-pression of NRF1 in the hippocampus was significantly upregulated following stereotactic injection of AAV-NRF1(P＜0.001).This intervention led to notable improvements in mitochondrial morphology within hippocampal neurons,as well as enhanced cognitive and memory functions in mice(P＜0.01).Moreover,there was a significant in-crease in dendritic spine density among neurons located in the CA1 region of the hippocampus(P＜0.001),ac-companied by long-lasting and stable long-term potentiation(LTP)and a substantial elevation in fEPSP slope(P＜0.01).Conclusion The overexpression of NRF1 in a 5 × FAD mouse model of Alzheimer's disease(AD)initia-ted the restoration of mitochondrial dysfunction and enhanced synaptic plasticity,indicating that these alterations may contribute to the therapeutic efficacy of NRF1 overexpression in ameliorating cognitive dysfunction associated with AD.
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Objective:To evaluate the effect of five-element acupuncture on clinical symptoms, brain metabolism and immunoglobulin level in patients with chronic fatigue syndrome.Methods:It was a randomized controlled trial. A total of 94 patients with chronic fatigue syndrome in our hospital from January 2021 to January 2022 were selected and divided into two groups according to the random number table method, with 47 in each group. The conventional western medicine group was treated with conventional western medicine, and the five-element acupuncture group was treated with five-element acupuncture on the basis of the conventional western medicine group. Both groups were treated for 4 weeks. Before and after treatment, the serum levels of interferon-γ (IFN-γ), corticosterone (CORT), IL-2 and 5-hydroxytryptamine (5-HT) were detected by ELISA; the levels of natural killer (NK) cells, CD4 +, CD8 +, IgG and IgM were detected by flow cytometry; the whole body superconducting MRI scanner was used to scan T2 Flair, T2WI and TlWI sequences of the hippocampus, and the spectral curves and the areas under the peak of N-acetylaspartic acid (NAA), creatine (Cr) and choline (Cho) were obtained, and the ratios of Cho/Cr and NAA/Cr were calculated. the fatigue Scale-14 (FS-14) and Fatigue Severity Scale (FSS) were used to evaluate the fatigue state of the patients, and the Depression-Anxiety-Stress Scale-21 (DASS-21) and Beck Anxiety Inventory (BAI) were used to evaluate the anxiety state of the patients. Symptom Checklist 90 (SCL-90) and Somatic and Mental Health Report Score (SPHERE) were used to evaluate the quality of life of patients. And the clinical efficacy was evaluated. Results:After treatment, the levels of IgG, CD4 + and NK in the five-element acupuncture group were significantly higher than those in the conventional western medicine group ( t values were 4.76, 3.65, 6.42, respectively, P<0.01), and the level of IgM, CD8 + was significantly lower than that in the conventional western medicine group ( t values were 7.30, 4.79, P<0.01); the levels of IFN-γ, IL-2 and 5-HT in the observation group were significantly higher than those in the conventional western medicine group ( t values were 7.60, 4.05, 2.79, respectively, respectively, P<0.01), and the level of CORT was significantly lower than that in the conventional western medicine group ( t=6.72, P<0.01); the NAA/Cr levels in the left [(1.10±0.04) vs. (1.05±0.03), t=6.86] and right [(1.18±0.02) vs. (1.21±0.03), t=8.23] hippocampus of the experimental group were significantly higher than those in the conventional western medicine group ( P<0.01), and the Cho/Cr levels in the left [(1.08±0.04) vs. (1.03±0.03), t=5.70] and right [(1.17±0.02) vs. (1.20±0.03), t=5.71] hippocampus of the experimental group were significantly lower than those of the conventional western medicine group ( P<0.01). After treatment, the scores of physical fatigue, mental fatigue and FSS in the five-element acupuncture group were significantly lower than those in the conventional western medicine group ( t values were 8.08, 9.08 and 7.07, respectively, P<0.01). The scores of DASS-21, BAI, SCL-90 and SPHERE in the conventional western medicine group were significantly lower than those in the conventional western medicine group ( t values were 3.63, 5.77, 8.74, 5.92, respectively, P<0.01).The total effective rate was 95.74% (45/47) in the five-element acupuncture group and 82.98% (39/47) in the conventional western medicine group, and there was no significant difference between the two groups ( χ2=2.80, P=0.094). Conclusion:Five-elements acupuncture can improve the expression of T lymphocytes, increase the levels of immunoglobulin and NK, reduce the level of CORT, regulate the brain metabolism of NAA in the left and right hippocampus, improve the clinical symptoms and negative emotions, and improve the clinical efficacy and quality of life in patients with chronic fatigue syndrome.
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ObjectiveTo investigate the effects of Lactobacillus rhamnosus GG （LGG）on microglia and Tau phosphorylation in the hippocampus of aged mice induced by anesthesia and surgery. MethodsA total of thirty 18-month-old C57BL/6J mice were randomly divided into three groups： control group， anesthesia surgery group， and anesthesia surgery + LGG group （10 mice/group）. The aged mice were oral administered by NS or LGG 109 CFU 150 μL once a day for 20 days. Then anesthesia surgery group and anesthesia surgery +LGG group received anesthesia with isoflurane and exploratory laparotomy. The activation status of microglia in the hippocampus was detected by immunofluorescence staining 12 hours after surgery. IL-6 concentration changes was detected by ELISA. The expression changes of Tau protein phosphorylation site （Tau-pS202/pT205） and total Tau protein was detected by western blot. ResultsThe microglia in the hippocampus of the control group were in a resting state， and the concentration of inflammatory factor IL-6 was （82.08 ± 12.07） pg/mL in control group. Compared to the control group， the anesthesia surgery group showed microglial cell Microglia were activated， the concentration of inflammatory factors IL-6 increased significantly to （123.7±5.72） pg/mL （P=0.000）， and the expression of phosphorylated Tau-pS202/pT205 increased the hippocampus （P=0.002）. Compared to the anesthesia surgery group， the activated microglia were inhibited， the concentration of IL-6 decreased to （96.68±9.59） pg/mL （P=0.008）， and the expression of phosphorylated Tau-pS202/pT205 reduced significantly in the AS+LGG group （P=0.002）. While there were no significant changes in total Tau protein among 3 groups. ConclusionPreoperative administration of probiotic LGG can alleviate the activation of microglia， increased secretion of inflammatory factors， and increased Tau protein phosphorylation levels in the hippocampus of elderly mice caused by anesthesia surgery.