Huannao Yicong Formula (还脑益聪方) Regulates γ-Secretase Activity through APH-1 and PEN-2 Gene Raguiation Pathways in Hippocampus of APP/PS1 Double Transgenic Mice

Objective: To observe the effects of Huannao Yicong Formula(还脑益聪方, HYF) on learning and memory and it's regulating effect on γ-secretase related anterior pharynx defective 1(APH-1), presenilin enhancer-2(PEN-2) signaling pathway, so as to discuss and further clarify the mechanism of HYF on Alzheimer's disease. Methods: Sixty APP/PS1 transgenic mice, randomly allocated into 4 groups, the model group, the donepezil group(0.65 mg/kg), HYF low-dose group(HYF-L, 5.46 g/kg) and HYF high-dose group(HYF-H, 10.92 g/kg), 15 for each group. Another 15 C57BL/6J mice with the same age and same genetic background were allocated into the control group, proper dosage of drugs or distilled water were given by intragastric administration once daily for 12 weeks. After 12 weeks of administration, the learning and memory abilities of mice in each group was evaluated by the morris water maze test, amyloid precursor protein(APP), Aβ1-40 and Aβ1-42 levels in hippocampus were detected by enzyme-linked immunosorbent assay, γ-secretase was detected by dual luciferase assaying, the levels of APH-1a, hypoxia-inducible factor 1α(HIF-1α), c AMP response element-binding protein(CREB) and PEN-2 and their m RNA expression was measured by Western blot and real-time polymerase chain reaction. Results: HYF can ameliorate learning and memory deficits in APP/PS1 transgenic mice by decreasing the escape latency, improving the number of platform crossing and swimming speed(P0.01, P0.05). HYF can decrease the levels of APP, Aβ1-40, Aβ1-42 and the activity of γ-secretase in hippocampus of Alzheimer's disease model mice. HYF can down-regulate the levels of CREB and PEN-2 and the expression of their m RNA. Conclusion: HYF can improve the learning and memory ability by inhibiting the activity of γ-secretase through the CREB/PEN-2 signaling pathway, and this may be one of the therapeutic mechanisms of HYF in Alzheimer's disease.     

Dendrobium Alkaloids Attenuates Learning and Memory Impairment in APP/PS1 Mice

Objective：To observe the effect of the total alkaloids of Dendrobium nobile Lindl on the learning and memory impairment of APP/PS1 transgenic mice. Methods：Seven months male APP/PS1 transgenic mice （n=24） were randomly divided into two groups：APP/PS1+vehicle and APP/ PS1+DNLA（the total alkaloids of Dendrobium nobile Lindl） groups. Age-matched male wild-type（ WT）littermates （n=24） were randomly divided into two groups：WT+vehicle and WT+DNLA groups. The normal group and APP / PS1 group were garaged with normal volume of saline（ NS） for 6 consecutive months. The mice in the normal administration group and APP/PS1-administered group were given the daily total alkaloid of Dendrobiumnobile 40 mg·kg-1. Morris water maze was used to detect learning and memory ability in mice. At the end of the behavioral test,the cortical area of senile plaques were detected in the mice by anesthesia,and the survival of the hippocampal neurons was detected by Nissl’s staining.Transmission electron microscope was performed to observe neuron structure and synaptic structure in hippocampus. The levels of IL-1β were measured by ELISA .The levels of Aβ1-40,Aβ1-42 and the expression of GFAP,IL-6,COX-2,p-NF-ΚB,p-p38,PSD95 and SYP in hippocampus were detected by Western blot. Results：Compared with WT+vehicle group,the mean escape latency was markedly increased and the time percentage in target quadrant showed notable decrease in APP/PS1+vehicle group. The number of neurons were significantly reduced in hippocampal CA1 region. The results of transmission electron microscope showed that the structure of neuron and synaptic structure were damaged and the number of synaptic density was decreased. The amyloid plaques,Aβ1-40,Aβ1-42 contents the protein expression of GFAP,IL-6,COX-2,p-NF-κB and p-p38 were increased,meanwhile,the protein expression of PSD95 and SYP were dramatically decreased in the hippocampus in APP/PS1+vehicle group. These effects in WT+DNLA group showed no notable differences compared to the WT+vehicle group. However,compared with APP/PS1+vehicle group,the mean escape latency was decreased and the time percentage in target quadrant was notably increased in APP/PS1+DNLA. Moreover,the number of neurons were significantly increased in hippocampal CA1 region. The
structure of neuron and synaptic structure was improvement Furthermore,the amyloid plaques,Aβ1-40,Aβ1-42 contents,GFAP,IL-6,COX-2,p- NF-κB and p-p38 expression were decreased in thehippocampus,the protein expression of PSD95 and SYP were significantly increased in APP/PS1+DNLA group. Conclusions：Under the experimental conditions,DNLA attenuates the learning and memory loss of Alzheimer’s disease mice and reduces the number of senile plaques and increases the number of surviving neurons. The mechanism may be related to level of Aβ,activation of astrocytes,activation of astrocytes,inhibition of NF-κB and p38 and improvement of synaptic dysfunction.     

Long-term impact of intrauterine MCMV infection on development of offspring nervous system

This study examined the impacts of intrauterine murine cytomegalovirus(MCMV) infection on the long-term learning and memory of offspring.Sexually matured male and female BALB/C mice without MCMV infection were identified by ELISA and then mated.Seventy pregnant mice were randomly divided into the virus group(n=40) and the control group(n=30),in which the pregnant mice were subjected to placenta inoculation of MCMV suspension(1 μL,1×106 PFU) or the same amount of cell culture medium,respectively,at gestational age of 12.5 days.Some pregnant mice [virus group(n=20),control group(n=15)] were sacrificed by cervical dislocation at gestational age of 18.5 days,and the head circumference and brain weight of the mouse fetuses were measured,and the MCMV infection in their brain tissues was detected by PCR.The other pregnant mice [virus group(n=20),control group(n=15)] delivered naturally,and the learning and memory capability of the offspring at 70-day-old was analyzed by Morris water maze test.The results showed that 28.57% mouse fetuses in the virus group developed viral infection in the brain.Their head circumference and brain weight were significantly reduced as compared with those in the control group(P<0.01).The Morris water maze test revealed that the mouse offspring in the control group found the platform with straight-line trajectories after training.In contrast,the counterparts in the virus group intended to enter the central area,but looked for the platform with a circular trajectory.And the infected mice exhibited prolonged swimming distance and swimming latency(P<0.01).It was concluded that:(1) placenta inoculation of MCMV can cause fetal brain infection and intrauterine development retardation;(2) the offspring of MCMV placenta inoculation mice showed a long-term decline in learning and memory capability.     

Angelica tenuissima Nakai Ameliorates Cognitive Impairment and Promotes Neurogenesis in Mouse Model of Alzheimer's Disease

Objective: To research Angelica tenuissima Nakai(ATN) for use in novel Alzheimer's disease(AD) therapeutics. Methods: The effect of a 30% ethanol extract of ATN(KH032) on AD-like cognitive impairment and neuropathological and neuroinflammatory changes induced by bilateral intracerebroventricular injections of β-amyloid(Aβ) peptide(Aβ_(1-42)) was investigated. Male C57 Bl/6 mice were randomly divided into 4 groups, 10 in each group. KH032-treated groups were administrated with a low or high dose of KH032(50 and 200 mg/kg, respectively), intragastrically for 16 days; distilled water was applied in the sham and negative groups. Open field test, Y maze and Morris water maze test were used for behavior test and cognitive ability. In addition, the neuroprotective effects of KH032 in Aβ_(1-42)-infused mice on the histopathological markers [neuronspecific nuclear protein(Neu N), Aβ1-42] of neurodegeneration were examined. The levels of glial fibrillary acidic protein(GFAP), Neu N, phosphorylation extracellular signal-regulated kinase(ERK)/ERK, brain-derived neurotrophic factor(BDNF), phosphorylation c AMP response element-binding(CREB)/CREB protein expression were measured by Western blot. Results: KH032 treatment ameliorated cognitive impairments, reduced the overexpression of Aβ_(1-42), and inhibited neuronal loss and neuroinflammatory response in the Aβ_(1-42)-infused mice. Moreover, KH032 treatment enhanced BDNF expression levels in the hippocampus. Finally, KH032 treatment increased phosphorylation of ERK1/2 and CREB, vital for ERK-CREB signaling. Conclusions: KH032 attenuated cognitive deficits in the Aβ_(1-42)-infused mice by increasing BDNF expression and ERK1/2 and CREB phosphorylation and inhibiting neuronal loss and neuroinflammatory response, suggesting that KH032 has therapeutic potential in neurodegenerative disorders such as AD.     

Effect and Safety of Huannao Yicong Formula(还脑益聪方) in Patients with Mild-to-Moderate Alzheimer's Disease: A Randomized,Double-Blinded,Donepezil-Controlled Trial

Objective: To assess the effect and safety of Huannao Yicong Formula(还脑益聪方, HYF) in the treatment of patients with mild-to-moderate Alzheimer's disease(AD). Methods: Sixty patients with mild-tomoderate AD were evenly randomized into HYF group and donepezil group with the random number method. Patients in the HYF group took 5 g of HYF granules twice daily and 5 mg placebo of donepezil once daily. Patients in the donepezil group took 5 mg donepezil once daily and 5 g placebo of HYF granules twice daily. The intervention lasted for 6 months. Clinical researchers, participants and statisticians were blinded to the treatment assignment throughout the study. The primary outcomes were scores of Alzheimer's Disease Assessment Scale-Cognitive Subscale(ADAS-Cog) and Chinese Medicine Symptom Scale(CM-SS). The secondary outcomes were scores of Montreal Cognitive Assessment(MoCA) test and Mini-Mental State Exam(MMSE). The serum levels of acetylcholinesterase(AchE) and amyloid-β protein 42(Aβ42) were detected with enzymelinked immunosorbent assay kits. The scale assessments were conducted at baseline, the 3 rd and 6 th months of treatment, respectively. Biochemistry tests were conducted at baseline and the 6 th month of treatment. Results: A total of 52 patients completed the trial, 28 in HYF group and 24 in donepezil group. Compared with the baseline, HYF and donepezil significantly decreased the total scores of ADAS-Cog and CM-SS, and significantly increased the scores of MoCA and MMSE after 6-month treatment(all P0.01). Both treatments remarkably reduced the serum levels of AchE and Aβ42(both P0.05). The CM-SS total effective rate of HYF was significantly higher than donepezil [75.00%(21/28) vs. 54.17%(13/24), P0.05]. No severe adverse events were observed in both groups. Conclusion: HYF is effective and safe for improving the cognitive function in mildto-moderate AD patients. [Trial registration: Chinese Clinical Trial Registry(Reg No. ChiCTR-IOR-17011746)]     

Overexpression of amyloid precursor protein inhibits neurite outgrowth and disrupts cytoskeleton in N2a cells

There is considerable evidence suggesting that altered metabolism of β-amyloid precursor protein (APP) and accumulation of its β-amyloid (Aβ) fragment are key features of Alzheimer's disease (AD). APP is a type Ⅰ integral membrane protein and consists of 695-770 amino acids encoded by differentially spliced mRNAs transcribed from a single gene located on human chromosome 21.1 The 695-amino acid APP is expressed preferentially in the brain. Aβ, the major component of senile plaques, is derived by proteolytic processing of APP by β-and γ-secretase and is constitutively released from most cells.     

Huannao Yicong Decoction(还脑益聪方) Extract Reduces Inflammation and Cell Apoptosis in Aβ_(1-42)-Induced Alzheimer's Disease Model of Rats

Objective: Huannao Yicong Decoction(还脑益聪方, HYD), an effective herbal formula against Alzheimer's disease(AD), has been proven to have neuroprotective action in amyloid β-protein_(1-42))(Aβ_(1-42))-induced rat model. This study was designed to characterize mechanisms by which HYD leads to suppression of inflammation and apoptosis in the brains of Aβ_(1-42)-induced rat. Methods: A total of 72 rats were divided into 6 groups, which were referred to as: sham operation group, model group, donepezil-treated group, HYD low-dose group(HYDL), HYD middle-dose group(HYDM) and HYD high-dose group(HYDH). Rats in HYDL, HYDM and HYDH were injected with Aβ_(1-42) at the CA1 region of hippocampus to form AD model and were fed the HYD extract at different dose of 3.78, 7.56 and 18.90 g crude drug/kg. The behavioral changes of rats were evaluated by Morris water maze(MWM) before sacrifice. Pathological changes of the brain tissue were evaluated using hematoxylin eosin(HE) staining. The levels of interleukin-1(IL-1) and tumor necrosis factor-α(TNF-α) were measured by radioimmunoassay. The levels of Aβ and proteins that are associated with apoptosis such as B-cell lymphoma-2 protein(Bcl-2), Bcl-2-associated X protein(Bax), cysteine-aspartic protease(caspase)-3,-8,-9 and-12 in serum were measured by immunohistochemistry. Results: Compared with the sham operation group, the spatial learning and memory abilities of AD rats were significantly decreased(P0.05 or P0.01; Expressions of IL-1, TNF-α, Aβ and apoptosis-signaling proteins caspase-3,-8,-9,-12 were significantly up-regulated(P0.05 or P0.01). The ratio of Bcl-2 to Bax were significantly decreased in the model group(P0.01). When treated with HYD extract, the spatial learning and memory abilities of AD-model rats were significantly increased(P0.05 or P0.01), IL-1, TNF-α, Aβ, caspase-3,-8,-9 and-12 were down-regulated(P0.05 or P0.01), and the ratio of Bcl-2 to Bax were reduced(P0.05 or P0.01). Conclusions: HYD extract can improve the learning and memory ability deficits, alleviate the inflammatory response and pathological manifestations induced by Aβ_(1-42) injection in the rat model of AD. HYD down-regulates the levels of IL-1, TNF-α and Aβ, and decreases the rate of apoptosis by modulating apoptosis-signaling-related proteins such as caspase-3,-8,-9, and-12.     

Neuroprotective Effect of Fructus broussonetiae on APP/PS1 Mice via Upregulation of AKT/β-Catenin Signaling

Objective:To evaluate the mechanisms underlying the protective effect of Chinese herbal medicine Fructus broussonetiae(FB)in both mouse and cell models of Alzheimer’s disease(AD).Methods:APP/PS1 mice treated with FB for 2 months and vehicle-treated controls were run through the Morris water maze and object recognition test to evaluate learning and memory capacity.RNA-Seq,Western blotting,and immunofluorescence staining were also conducted to evaluate the effects of FB treatment on various signaling pathways altered in APP/PS1 mice.To further explore the mechanisms underlying FB’s protective effect,PC-12 cells were treated with Aβ25–35 in order to establish an in vitro model of AD.Results:FB-treated mice showed improved learning and memory capacity on both the Morris water maze and object recognition tests.RNA-seq of hippocampal tissue from APP/PS1 mice showed that FB had effects on multiple signaling pathways,specifically decreasing cell apoptotic signaling and increasing AKT and β-catenin signaling.Similarly,FB up-regulated both AKT and β-catenin signaling in PC-12 cells pre-treated with Aβ25–35,in which AKT positively regulated β-catenin signaling.Further study showed that AKT promoted β-catenin signaling via enhancing β-catenin(Ser552)phosphorylation.Moreover,AKT and β-catenin signaling inhibition both resulted in the attenuated survival of FB-treated cells,indicating the AKT/β-catenin signaling is a crucial mediator in FB promoted cell survival.Conclusions:FB exerted neuroprotective effects on hippocampal cells of APP/PS1 mice,as well as improved cell viability in an in vitro model of AD.The protective actions of FB occurred via the upregulation of AKT/β-catenin signaling.     

Influence of High Level TGF-β1 on Scleral Thickness

In order to explore the role of TGF-β1 in scleral remodeling and the possible mechanism,the influence of high level TGF-β1 on scleral thickness and the expression of MMP-2 and TIMP-2 was investigated in a TGF-β1 transgenic mouse model.Alb/TGF-β1(Cys223,225Ser) TGF-β1 transgenic mice were used as experimental subjects and non-transgenic littermates as controls.Plasma levels of TGF-β1 were determined by ELISA.TGF-β1,MMP-2 and TIMP-2 levels in sclera were detected by using Western blot.The thickness of posterior sclera was measured by computerized image analysis of a midsagittal section.Mean difference was analyzed with independent t-test.The results showed plasma levels of TGF-β1 in transgenic mice were 1.68 times as much as that in the controls(P<0.01).TGF-β1 levels in the sclera of transgenic mice were 2.68 times of the controls(P<0.01).Posterior scleral thickness in transgenic mice were significantly thicker than in the controls.There was no significant difference in the MMP-2 levels between transgenic mice and controls,but the TIMP-2 levels were increased significantly in transsgenic mice as compared with those in the controls.It was suggested that high levels of TGF-β1 in transgenic mice could result in the increased scleral thickness by inducing the expression of TIMP-2 to suppress the activity of MMP-2,finally inhibiting the degradation of collagen.     

Potential association of lead exposure during early development of mice with alteration of hippocampus nitric oxide levels and learning memory

Objective Chronic lead （Pb） exposure during development is known to produce learning deficits. Nitric oxide participates in the synaptic mechanisms involved in certain forms of learning and memory. This study was designed to clarify whether Pb-induced impairment in learning and memory was associated with the changes of nitric oxide levels in mice brains. Methods Sixty Balb/c mice aged l0 days were chosen. A model of lead exposure was established by drinking 0.025%, 0.05% 0.075% lead acetate, respectively for 8 weeks. The controls were orally given distilled water. The ability to learn and memorize was examined by open field test, T-water maze test. In parallel with the behavioral data, NO level of hippocampus tissue was detected by biochemical assay. Results Compared with control groups, （1） the weight of 0.075% group was significantly reduced （P〈0.05）; （2） The number of times in mice attaining the required standards in T-water maze test was lower in 0.075% group （P〈0.01）. No significant difference was found between experimental and control groups in open field test （P〉0.05）; （3） NO level of mouse hippocampus tissue was decreased in 0.075% group （P〈0.01）. Conclusions The findings suggest that decreased hippocampus NO level may contribute to the Pb-induced deficits in learning and memory processes.     

Protective Effect of Mulberry Extract against Pb-induced Learning and Memory Deficits in Mice

<正>Lead(Pb)is ubiquitous in the environment,and low-level Pb exposure can cause neurotoxicity and irreversible damage to children’s cognition,learning and memory ability.Nutritional intervention is an effective method to prevent Pb poisoning.Mulberry is rich in anthocyanins,possessing protective effects for nerves.This study investigated the neuroprotective effects of mulberry extract(ME)against Pb-induced learning and memory deficits in mice.The results showed that the learning and memory abilities of mice,assessed using the Morris test,improved significantly after treatment with ME at a dose of 100 mg/kg body weight.The level of Pb in the brains of mice in the three ME intervention groups decreased significantly,while NO production and anti-oxidant enzymes were significantly restored.It is suggested that ME inhibits Pb-induced neurotoxicity by reversing Pb-induced alterations in the aspect of neurotoxic effects and improving learning and memory.     

Huannao Yicong Decoction (还脑益聪方) extract reduces inflammation and cell apoptosis in Aβ1-42-induced Alzheimer's disease model of rats

Objective: Huannao Yicong Decoction (还脑益聪方, HYD), an effective herbal formula against Alzheimer''s disease (AD), has been proven to have neuroprotective action in amyloid β-protein1-42 (Aβ1-42)-induced rat model. This study was designed to characterize mechanisms by which HYD leads to suppression of inflammation and apoptosis in the brains of Aβ1-42-induced rat. Methods: A total of 72 rats were divided into 6 groups, which were referred to as: sham operation group, model group, donepezil-treated group, HYD low-dose group (HYDL), HYD middle-dose group (HYDM) and HYD high-dose group (HYDH). Rats in HYDL, HYDM and HYDH were injected with Aβ1-42 at the CA1 region of hippocampus to form AD model and were fed the HYD extract at different dose of 3.78, 7.56 and 18.90 g crude drug/kg. The behavioral changes of rats were evaluated by Morris water maze (MWM) before sacrifice. Pathological changes of the brain tissue were evaluated using hematoxylin eosin (HE) staining. The levels of interleukin-1 (IL-1) and tumor necrosis factor-α (TNF-α) were measured by radioimmunoassay. The levels of Aβ and proteins that are associated with apoptosis such as B-cell lymphoma-2 protein (Bcl-2), Bcl-2-associated X protein (Bax), cysteine-aspartic protease (caspase)-3, -8, -9 and -12 in serum were measured by immunohistochemistry. Results: Compared with the sham operation group, the spatial learning and memory abilities of AD rats were significantly decreased (P<0.05 or P<0.01; Expressions of IL-1, TNF-α, Aβ and apoptosis-signaling proteins caspase-3, -8, -9, -12 were significantly up-regulated (P<0.05 or P<0.01). The ratio of Bcl-2 to Bax were significantly decreased in the model group (P<0.01). When treated with HYD extract, the spatial learning and memory abilities of AD-model rats were significantly increased (P<0.05 or P<0.01), IL-1, TNF-α, Aβ, caspase-3, -8, -9 and -12 were down-regulated (P<0.05 or P<0.01), and the ratio of Bcl-2 to Bax were reduced (P<0.05 or P<0.01). Conclusions: HYD extract can improve the learning and memory ability deficits, alleviate the inflammatory response and pathological manifestations induced by Aβ1-42 injection in the rat model of AD. HYD down-regulates the levels of IL-1, TNF-α and Aβ, and decreases the rate of apoptosis by modulating apoptosis-signaling-related proteins such as caspase-3, -8, -9, and -12.     

Angelica tenuissima Nakai Ameliorates Cognitive Impairment and Promotes Neurogenesis in Mouse Model of Alzheimer's Disease

Objective: To research Angelica tenuissima Nakai (ATN) for use in novel Alzheimer''s disease (AD) therapeutics. Methods: The effect of a 30% ethanol extract of ATN (KH032) on AD-like cognitive impairment and neuropathological and neuroinflammatory changes induced by bilateral intracerebroventricular injections of β-amyloid (Aβ) peptide (Aβ1-42) was investigated. Male C57Bl/6 mice were randomly divided into 4 groups, 10 in each group. KH032-treated groups were administrated with a low or high dose of KH032 (50 and 200 mg/kg, respectively), intragastrically for 16 days; distilled water was applied in the sham and negative groups. Open field test, Y maze and Morris water maze test were used for behavior test and cognitive ability. In addition, the neuroprotective effects of KH032 in Aβ1-42-infused mice on the histopathological markers [neuron-specific nuclear protein (NeuN), Aβ1-42] of neurodegeneration were examined. The levels of glial fibrillary acidic protein (GFAP), NeuN, phosphorylation extracellular signal-regulated kinase (ERK)/ERK, brain-derived neurotrophic factor (BDNF), phosphorylation cAMP response element-binding (CREB)/CREB protein expression were measured by Western blot. Results: KH032 treatment ameliorated cognitive impairments, reduced the overexpression of Aβ1-42, and inhibited neuronal loss and neuroinflammatory response in the Aβ1-42-infused mice. Moreover, KH032 treatment enhanced BDNF expression levels in the hippocampus. Finally, KH032 treatment increased phosphorylation of ERK1/2 and CREB, vital for ERK-CREB signaling. Conclusions: KH032 attenuated cognitive deficits in the Aβ1-42-infused mice by increasing BDNF expression and ERK1/2 and CREB phosphorylation and inhibiting neuronal loss and neuroinflammatory response, suggesting that KH032 has therapeutic potential in neurodegenerative disorders such as AD.     

Behavioral Abnormality along with NMDAR-related CREB Suppression in Rat Hippocampus after Shortwave Exposure

Objective To estimate the detrimental effects of shortwave exposure on rat hippocampal structure and function and explore the underlying mechanisms. Methods One hundred Wistar rats were randomly divided into four groups(25 rats per group) and exposed to 27 MHz continuous shortwave at a power density of 5, 10, or 30 m W/cm^2 for 6 min once only or underwent sham exposure for the control. The spatial learning and memory, electroencephalogram(EEG), hippocampal structure and Nissl bodies were analysed. Furthermore, the expressions of N-methyl-D-aspartate receptor(NMDAR) subunits(NR1, NR2 A, and NR2 B), c AMP responsive element-binding protein(CREB) and phosphorylated CREB(p-CREB) in hippocampal tissue were analysed on 1, 7, and 14 days after exposure. Results The rats in the 10 and 30 m W/cm^2 groups had poor learning and memory, disrupted EEG oscillations, and injured hippocampal structures, including hippocampal neurons degeneration, mitochondria cavitation and blood capillaries swelling. The Nissl body content was also reduced in the exposure groups. Moreover, the hippocampal tissue in the 30 m W/cm^2 group had increased expressions of NR2 A and NR2 B and decreased levels of CREB and p-CREB. Conclusion Shortwave exposure(27 MHz, with an average power density of 10 and 30 m W/cm^2) impaired rats' spatial learning and memory and caused a series of dose-dependent pathophysiological changes. Moreover, NMDAR-related CREB pathway suppression might be involved in shortwave-induced structural and functional impairments in the rat hippocampus.     

Hawthorn Extract Alleviates Atherosclerosis through Regulating Inflammation and Apoptosis Related Factors: An Experimental Study

Objective: To determine the effects of hawthorn extract on serum lipid levels, pathological changes in aortic atherosclerosis plaque, inflammatory factors, and apoptosis-related protein and mRNA expression in apolipoprotein E gene knockout(ApoE~(-/-)) mice. Methods: Thirty-six ApoE~(-/-) mice were fed with a high-fat diet starting at the age of 8 weeks. Mice were randomly divided into 3 groups by a random number table including model group, hawthorn extract group, and simvastatin group, 12 mice in each group. Twelve 8-week-old C57BL/6 mice were fed a basic diet and served as control. The mice in the control and model groups were administered 0.2 mL saline daily, the mice in the hawthorn extract and simvastatin groups were administered with 50 mg/kg hawthorn extract or 5 mg/kg simvastatin daily for 16 weeks. After 16 weeks, plasma lipids including total cholesterol(TC), triglyceride(TG), low-density lipoprotein cholesterol(LDL-C) and high-density lipoprotein cholesterol(HDL-C) were determined by an enzymatic assay. Aortic atherosclerotic lesions were observed by light microscopy, scanning and transmission electron microscopy, respectively. Plasma levels of monocyte chemoattractant protein-1(MCP-1), interleukin-1β(IL-1β), adiponectin(APN), and hypersensitive C-reactive protein(hs-CRP) were measured by enzyme-linked immunosorbent assay(ELISA). Protein and mRNA expressions of Bax and Bcl-2 in the aorta were assessed by Western blotting and quantitative real-time polymerase chain reaction(qR T-PCR), respectively. Results: Compared to the control group, the plasma levels of TC, TG and LDL-C were significantly increased and HDL-C were significantly decreased in the model group(P0.01). Compared to the model group, treatment with hawthorn extract significantly decreased the plasma levels of TC, TG, and LDL-C and increased the plasma level of HDL-C in ApoE~(-/-)mice(P0.01). The levels of MCP-1, IL-1β, and hs-CRP in the model group were significantly increased and APN was significantly decreased compared with the control group(P0.01). Compared to the model group, treatment with hawthorn extract decreased the levels of MCP-1, IL-1β, and hs-CRP and increased the APN level(P0.01). Compared to the control group, the protein and mR NA expression of Bax in the model group were significantly increased and the expression of Bcl-2 was significantly decreased(P0.01). Hawthorn extract also reduced the protein and mR NA expression of Bax and increased the Bcl-2 expression in the aorta(P0.01). Conclusion: Hawthorn extract has anti-atherosclerosis and stabilizing unstable plaque effects. The mechanism may be related to the inflammation and apoptosis signaling pathways.     

Heijiangdan Ointment(黑绛丹膏)Relieves Oxidative Stress from Radiation Dermatitis Induced by ~(60)Co γ-Ray in Mice

Objective:To investigate the effects of Heijiangdan Ointment(黑绛丹膏,HJD) on oxidative stress in ~(60)Co γ-ray radiation-induced dermatitis in mice.Methods:Female Wistar mice with grade 4 radiation dermatitis induced by ~(60)Co γ-rays were randomly divided into four groups(n=12 per group);the HJD-treated,recombinant human epidermal growth factor(rhEGF)-treated,Trolox-treated,and untreated groups,along with a negative control group.On the 11 th and 21 st days after treatment,6 mice in each group were chosen for evaluation.The levels of superoxide dismutase(SOD),malondialdehyde(MDA),and lactate dehydrogenase(LDH) were detected using spectrophotometric methods.The fibroblast mitochondria were observed by transmission electron microscopy(TEM).The expressions of fibroblast growth factor 2(FGF-2) and transforming growth factor β1(TGF-β1) were analyzed by western blot.Results:Compared with the untreated group,the levels of SOD,MDA and LDH,on the 11 th and 21 st days after treatment showed significant difference(P0.05).TEM analysis indicated that fibroblast mitochondria in the untreated group exhibited swelling and the cristae appeared fractured,while in the HJD group,the swelling of mitochondria was limited and the rough endoplasmic reticulum appeared more relaxed.The expressions of FGF-2 and TGF-β1 increased in the untreated group compared with the negative control group(P0.05).After treatment,the expression of FGF-2,rhEGF and Trolox in the HJD group were significantly increased compared with the untreated group(P0.05),or compared with the negative control group(P0.05).The expression of TGF-β1 showed significant difference between untreated and negative control groups(P0.05).HJD and Trolox increased the level of TGF-β1 and the difference was marked as compared with the untreated and negative control groups(P0.05).Conclusion:HJD relieves oxidative stress-induced injury,increases the antioxidant activity,mitigates the fibroblast mitochondrial damage,up-regulates the expression of growth factor,and promotes mitochondrial repair in mice.     

The Effect of Dendrobium Nobile Lindl. Alkaloids on the Protein Expression of Aβ42，APP and BACE1 of Hippocampus in db/db Mice

Objective：Beta-amyloid （Aβ） deposition is considered as vital factor leading to cognitive impairment in Alzheimer’s disease（ AD）.In addition,there are pathophysiological connections between Type 2 Diabetes Mellitus （T2DM） and AD. Diabetic patients have higher incidences of cognitive impairment and hence they are more at the risk of developing AD. As one of the active compounds of Dendrobium nobile Lindl. Dendrobium nobile Lindl. Alkaloids （DNLA） has the effect of protecting the nervous system and decreased the level of fasting blood glucose （FBG） in T2DM model mice. In this study,we attempted to investigate effects of DNLA on the proteins expression of APP,BACE1 and Aβ42 of hippocampus in db/db mice. Methods：10 male C57BL/KsJ mice were
control group. And 4-week-old mice male db/db mice were randomly divided into four groups：model,DNLA-L（ 20 mg·kg-1）,DNLA-M（ 40 mg·kg-1）,and DNLA-H （80 mg·kg-1）,there are 9 mice in each group. After mice were treated with different concentration DNLA by gavaged for 17 weeks. The protein expression of β-amyloid 42 （Aβ42）,β-site amyloid precursor protein-cleaving enzyme 1 （BACE1） and amyloid precursor protein （APP） were examined by Western Blotting. Results：Compared with control group,not only the protein expression of Aβ42,but also BACE1 and APP was signifi cantly increased in hippocampus of model group. Moreover,DNLA signifi cantly decreased the protein expressions of Aβ42,BACE1 and APP in hippocampus of db/db mice compared with model group,and decreased in a dosedependent manner. Conclusion：DNLA can decrease the protein expressions of Aβ42 in hippocampus of db/ db mice. And the mechanism may be involved the decrease of BACE1 and APP.     

Effects of chronic multiple stress on learning and memory and the expression of Fyn, BDNF, TrkB in the hippocampus of rats

Background The effect of chronic stress on cognitive functions has been one of the hot topics in neuroscience. But there has been much controversy over its mechanism. The aim of this study was to investigate the effects of chronic multiple stress on spatial learning and memory as well as the expression of Fyn, BDNF and TrkB in the hippocampus of rats. Methods Adult rats were randomly divided into control and chronic multiple stressed groups. Rats in the multiple stressed group were irregularly and alternatively exposed to situations of vertical revolution, sleep expropriation and restraint lasting for 6 weeks, 6 hours per day with night illumination for 6 weeks. Before and after the period of chronic multiple stresses, the performance of spatial learning and memory of all rats was measured using the Morris Water Maze （MWM）. The expression of Fyn, BDNF and TrkB proteins in the hippocampus was assayed by Western blotting and immunohistochemical methods. The levels of Fyn and TrkB mRNAs in the hippocampus of rats were detected by RT-PCR technique. Results The escape latency in the control group and the stressed group were 15.63 and 8.27 seconds respectively. The performance of spatial learning and memory of rats was increased in chronic multiple stressed group （P〈0.05）. The levels of Fyn, BDNF and TrkB proteins in the stressed group were higher than those of the control group （P〈0.05）. The results of immunoreactivity showed that Fyn was present in the CA3 region of the hippocampus and BDNF positive particles were distributed in the nuclei of CA1 and CA3 pyramidal cells as well as DG granular cells. Quantitative analysis indicated that level of Fyn mRNA was also upregulated in the hippocampus of the stressed group （P〈0.05）. Conclusions Chronic multiple stress can enhance spatial learning and memory function of rats. The expression of Fyn, BDNF and TrkB proteins and the level of Fyn mRNA are increased in the stessed rat hippocampus. These suggest that Fyn and BDNF/TrkB signal transduction pathways may participate in the process of the enhanced learning and memory durina chronic multiple stress.