Alleviative effect of scopolamine­induced memory deficit via enhancing antioxidant and cholinergic function in rats by pinostrobin from Boesenbergia rotunda (L.).

Pinostrobin, a key bioactive compound found in the medicinal plant Boesenbergia rotunda (L.), has been noted for its beneficial biological properties including antioxidant, anti-inflammation, anti-cancer and anti-amnesia activities. In view of this, the present study purposed to evaluate the neuroprotective potential of pinostrobin in reversing scopolamine-induced cognitive impairment involving oxidative stress and cholinergic function in rats. A total of 30 male Wistar rats were randomly divided into five groups (n=6): Group 1 received vehicle as a control, group 2 received vehicle + scopolamine (3 mg/kg, i.p.), group 3 received pinostrobin (20 mg/kg, p.o.) + scopolamine, group 4 received pinostrobin (40 mg/kg, p.o.) + scopolamine and group 5 received donepezil (5 mg/kg, p.o.) + scopolamine. Treatments were administered orally to the rats for 14 days. During the final 7 days of treatment, a daily injection of scopolamine was administered. Scopolamine impaired learning and memory performance, as measured by the novel object recognition test and the Y-maze test. Additionally, oxidative stress marker levels, acetylcholinesterase (AChE) activity, choline acetyltransferase (ChAT) and glutamate receptor 1 (GluR1) expression were determined. Consequently, the findings demonstrated that the administration of pinostrobin (20 and 40 mg/kg) markedly improved cognitive function as indicated by an increase in recognition index and by spontaneous alternation behaviour. Pinostrobin also modulated the levels of oxidative stress by causing a decrease in malondialdehyde levels accompanied by increases in superoxide dismutase and glutathione activities. Similarly, pinostrobin markedly enhanced cholinergic function by decreasing AChE activity and promoting ChAT immunoreactivity in the hippocampus. Additionally, the reduction in GluR1 expression due to scopolamine was diminished by treatment with pinostrobin. The findings indicated that pinostrobin exhibited a significant restoration of scopolamine-induced memory impairment by regulating oxidative stress and cholinergic system function. Thus, pinostrobin could serve as a potential therapeutic agent for the management of neurodegenerative diseases such as Alzheimer's disease.

Role of acetylcholine and acetylcholinesterase in improving abiotic stress resistance/tolerance.

Abiotic stress that plants face may impact their growth and limit their productivity. In response to abiotic stress, several endogenous survival mechanisms get activated, including the synthesis of quaternary amines in plants. Acetylcholine (ACh), a well-known quaternary amine, and its components associated with cholinergic signaling are known to contribute to a variety of physiological functions. However, their role under abiotic stress is not well documented. Even after several studies, there is a lack of a comprehensive understanding of how cholinergic components mitigate abiotic stress in plants. Acetylcholine hydrolyzing enzyme acetylcholinesterase (AChE) belongs to the GDSL lipase/acylhydrolase protein family and has been found in several plant species. Several studies have demonstrated that GDSL members are involved in growth, development, and abiotic stress. This review summarizes all the possible mitigating effects of the ACh-AChE system on abiotic stress tolerance and will try to highlight all the progress made so far in this field.

Nasal solitary chemosensory cells govern daily rhythm in mouse model of allergic rhinitis.

BACKGROUND: While the daily rhythm of allergic rhinitis (AR) has long been recognized, the molecular mechanism underlying this phenomenon remains enigmatic. OBJECTIVE: We aimed to investigate the role of circadian clock in AR development and to clarify the mechanism by which the daily rhythm of AR is generated. METHODS: AR was induced in mice with ovalbumin. Toluidine blue staining, liquid chromatography-tandem mass spectrometry analysis, real-time quantitative PCR, and immunoblotting were performed with AR and control mice. RESULTS: Ovalbumin-induced AR is diurnally rhythmic and associated with clock gene disruption in nasal mucosa. In particular, Rev-erbα is generally downregulated and its rhythm retained, but with a near-12-hour phase shift. Furthermore, global knockout of core clock gene Bmal1 or Rev-erbα increases the susceptibility of mice to AR and blunts AR rhythmicity. Importantly, nasal solitary chemosensory cells (SCCs) are rhythmically activated, and inhibition of the SCC pathway leads to attenuated AR and a loss of its rhythm. Moreover, rhythmic activation of SCCs is accounted for by diurnal expression of ChAT (an enzyme responsible for the synthesis of acetylcholine) and temporal generation of the neurotransmitter acetylcholine. Mechanistically, Rev-erbα trans-represses Chat through direct binding to a specific response element, generating a diurnal oscillation in this target gene. CONCLUSION: SCCs, under the control of Rev-erbα, are a driver of AR rhythmicity; targeting SCCs should be considered as a new avenue for AR management.

Prebiotics counteract the morphological and functional changes secondary to chronic cisplatin exposition in the proximal colon of mice.

Cisplatin is an antimitotic drug able to cause acute and chronic gastrointestinal side effects. Acute side effects are attributable to mucositis while chronic ones are due to neuropathy. Cisplatin has also antibiotic properties inducing dysbiosis which enhances the inflammatory response, worsening local damage. Thus, a treatment aimed at protecting the microbiota could prevent or reduce the toxicity of chemotherapy. Furthermore, since a healthy microbiota enhances the effects of some chemotherapeutic drugs, prebiotics could also improve this drug effectiveness. We investigated whether chronic cisplatin administration determined morphological and functional alterations in mouse proximal colon and whether a diet enriched in prebiotics had protective effects. The results showed that cisplatin caused lack of weight gain, increase in kaolin intake, decrease in stool production and mucus secretion. Prebiotics prevented increases in kaolin intake, changes in stool production and mucus secretion, but had no effect on the lack of weight gain. Moreover, cisplatin determined a reduction in amplitude of spontaneous muscular contractions and of Connexin (Cx)43 expression in the interstitial cells of Cajal, changes that were partially prevented by prebiotics. In conclusion, the present study shows that daily administration of prebiotics, likely protecting the microbiota, prevents most of the colonic cisplatin-induced alterations.

Apparent absence of hypothalamic cholinergic neurons in the common ostrich and emu: Implications for global brain states during sleep.

We examined the presence/absence and parcellation of cholinergic neurons in the hypothalami of five birds: a Congo grey parrot (Psittacus erithacus), a Timneh grey parrot (P. timneh), a pied crow (Corvus albus), a common ostrich (Struthio camelus), and an emu (Dromaius novaehollandiae). Using immunohistochemistry to an antibody raised against the enzyme choline acetyltransferase, hypothalamic cholinergic neurons were observed in six distinct clusters in the medial, lateral, and ventral hypothalamus in the parrots and crow, similar to prior observations made in the pigeon. The expression of cholinergic nuclei was most prominent in the Congo grey parrot, both in the medial and lateral hypothalamus. In contrast, no evidence of cholinergic neurons in the hypothalami of either the ostrich or emu was found. It is known that the expression of sleep states in the ostrich is unusual and resembles that observed in the monotremes that also lack hypothalamic cholinergic neurons. It has been proposed that the cholinergic system acts globally to produce and maintain brain states, such as those of arousal and rapid-eye-movement sleep. The hiatus in the cholinergic system of the ostrich, due to the lack of hypothalamic cholinergic neurons, may explain, in part, the unusual expression of sleep states in this species. These comparative anatomical and sleep studies provide supportive evidence for global cholinergic actions and may provide an important framework for our understanding of one broad function of the cholinergic system and possible dysfunctions associated with global cholinergic neural activity.

Adeno-Associated Virus Type 9-Mediated Gene Therapy of Choline Acetyltransferase-Deficient Mice.

The enzyme choline acetyltransferase (ChAT) synthesizes acetylcholine from acetyl-CoA and choline at the neuromuscular junction and at the nerve terminals of cholinergic neurons. Mutations in the ChAT gene (CHAT) result in a presynaptic congenital myasthenic syndrome (CMS) that often associates with life-threatening episodes of apnea. Knockout mice for Chat (Chat-/-) die at birth. To circumvent the lethality of this model, we crossed mutant mice possessing loxP sites flanking Chat exons 4 and 5 with mice that expressed Cre-ERT2. Injection of tamoxifen (Tx) at postnatal (P) day 11 in these mice induced downregulation of Chat, autonomic failure, weakness, and death. However, a proportion of Chatflox/flox-Cre-ERT2 mice receiving at birth an intracerebroventricular injection of 2 × 1013 vg/kg adeno-associated virus type 9 (AAV9) carrying human CHAT (AAV9-CHAT) survived a subsequent Tx injection and lived to adulthood without showing signs of weakness. Likewise, injection of AA9-CHAT by intracisternal injection at P28 after the onset of weakness also resulted in survival to adulthood. The expression of Chat in spinal motor neurons of Chatflox/flox-Cre-ERT2 mice injected with Tx was markedly reduced, but AAV-injected mice showed a robust recovery of ChAT expression, which was mainly translated by the human CHAT RNA. The biodistribution of the viral genome was widespread but maximal in the spinal cord and brain of AAV-injected mice. No significant histopathological changes were observed in the brain, liver, and heart of AAV-injected mice after 1 year follow-up. Thus, AAV9-mediated gene therapy may provide an effective and safe treatment for patients severely affected with CHAT-CMS.

Distribution of alpha-synuclein in rat salivary glands.

Expression of alpha-synuclein (Syn), a presynaptic neuronal protein, was immunohistochemically examined in intact rat submandibular, sublingual, and lingual glands. The submandibular gland contained abundant periductal Syn-immunoreactive (-ir) nerve fibers. Abundant Syn-ir varicosities were present in acini of the sublingual and serous lingual glands. By confocal laser scanning microscopy, Syn-ir nerve fibers around smooth muscle actin (SMA)-ir cells alone were infrequent; however, those around aquaporin-5 (AQP5)-ir cells alone and both SMA- and AQP5-ir cells were abundant in the sublingual and serous lingual glands. SMA-ir cells were occasionally immunoreactive for toll-like receptor 4, a Syn receptor. Syn-ir nerve fibers contained tyrosine hydroxylase (TH) in the submandibular gland and choline acetyltransferase (ChAT) in all examined salivary glands. In the superior cervical (SCG), submandibular, and intralingual ganglia, sympathetic and parasympathetic neurons co-expressed Syn with TH and ChAT, respectively. SCG neurons innervating the submandibular gland contained mostly Syn. In the thoracic spinal cord, 14.7% of ChAT-ir preganglionic sympathetic neurons co-expressed Syn. In the superior salivatory nucleus, preganglionic parasympathetic neurons projecting to the lingual nerve co-expressed Syn and ChAT. The present findings indicate that released Syn acts on myoepithelial cells. Syn in pre- and post-ganglionic neurons may regulate neurotransmitter release and salivary volume and composition.

Effect of human urinary kallidinogenase on cognitive function of SAMP8 mouse model and its mechanism / 中国药理学通报

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.

Effect of Ultrafine Powder of Gastrodiae Rhizoma in Improving Learning and Memory Ability of Vascular Dementia Rats by Regulating Cholinergic System / 中国实验方剂学杂志

Objective:To investigate whether ultrafine powder of Gastrodiae Rhizoma （UPG） can alleviate the learning and memory impairment of vascular dementia rats and delay the process of VD， and whether this effect is related to the release of acetylcholine （Ach） through the regulation with acetylcholinesterase （AChE） and choline acetyltransferase （ChAT） and control of cholinergic system. Method:SD rats were randomly divided into sham operation group， UPG low dose group （0.45 g·kg-1）， UPG high dose group （1.8 g·kg-1） and Huperzine A group （80 μg·kg-1）， with 12 rats in each group. The drug administration groups were given orally drugs once a day for 8 weeks， and sham group and model group were given orally the same amount of distilled water. The learning and memory ability of the rats with VD were evaluated by the Morris water maze. Htoxylin eosin（HE） staining was used for pathomorphological observation of hippocampus CA1 area of the rats. The content of Ach was determined by enzyme-linked immunosorbent assay（ELISA）， AChE and ChAT protein expressions were detected by Western blot， and expression of ChAT in hippocampus CA1 area was observed by immunohistochemistry. Result:Compared with the sham operation group， the escape latency of the model group was significantly increased （P<0.01）， and the frequency of crossings platform and the time of staying in the target quadrant were reduced significantly （P<0.01）. HE staining of hippocampal tissues from VD rat showed neuron disorders， loss and degeneration and necrosis， pyknosis of the nucleus and light coloration of the cytoplasm. The level of acetylcholine in the hippocampus was significantly decreased by ELISA （P<0.05）， the expression level of AChE protein was significantly up-regulated， and the expression level of ChAT protein was significantly down-regulated （P<0.01）. Compared with model group， each administration group could significantly reduce the escape latency of the model rats， and significantly increase the frequency of crossing platform and the time of staying in the target quadrant （P<0.01）， the content of Ach was significantly increased （P<0.05）， the expression of AChE protein was significantly down-regulated （P<0.01）， while the expression of ChAT protein was significantly up-regulated （P<0.01）. Conclusion:UPG improves the learning and memory ability of vascular dementia rats， and its mechanism may be related to the increase of Ach， ChAT level and the decrease of AChE level.

Effects of Shenqi Xingnao Prescription on Learning and Memory Ability, Contents of Choline Acetyltransferase and Acetylcholine Esterase in Brain Tissue of Mice Models with Alzheimer Disease / 中国中医药信息杂志

Objective To observe the effects of Shenqi Xingnao Prescription on learning and memory ability, contents of choline acetyltransferase (ChAT) and acetylcholine esterase (AChE) in brain tissue in mice models with scopolamine-induced Alzheimer disease (AD); To investigate its mechanism for prevention and treatment for AD. Methods Totally 110 ICR mice were randomly divided into control group, control+Shenqi Xingnao Prescription high-dose group,model group,donepezil group,model+Shenqi Xingnao Prescription high-,medium-,and low-dose groups. The control and model group were given distilled water for gavage, and the other groups were given the corresponding medicine for gavage, once a day, for 14 days. On the 15th day, Morris water maze test and object recognition test were used to evaluate the learning and memory ability of each group. The model mice of memory impairment induced by intraperitoneal injection of scopolamine was established 20 minutes before the behavioral test. The expressions of ChAT and AChE in cortex and hippocampus were detected by Western blot. Results The results of Morris water maze test showed that compared with the control group, the model group had significant longer escape latency(P<0.05);Compared with the model group,Shenqi Xingnao Prescription medium-and high-dose groups could shorten the escape latency (P<0.05). The results of the object recognition test showed that compared with the control group, the ability of the model group to explore new things decreased and the discrimination index (DI) decreased (P<0.001);Compared with the model group,Shenqi Xingnao Prescription groups could increase the DI of model mice (P<0.05, P<0.01, P<0.001). The results of Western blot showed that the expression of AChE protein in the cortex and hippocampus of the model group was significantly higher than the control group (P<0.01); Compared with the model group, Shenqi Xingnao Prescription low- and medium-dose groups could decrease the expression of AChE in the cortex in different degrees(P<0.01);Shenqi Xingnao Prescription groups could decreaed the expression of AChE in the hippocampus (P<0.001); There was no significant statistical significance in the expression of ChAT in the cortex and hippocampus in each group.Conclusion Shenqi Xingnao Prescription can significantly improve the learning and memory ability of AD model mice induced by scopolamine, which may be related to the descent expression of AChE protein in the cortex and hippocampus of the model mice.

Effects of Cornel Iridoid Glycoside on Learning-memory Ability and Brain Pathological Changes in Vascular Dementia Rats / 中国中医药信息杂志

Objective To investigate the effects of cornel iridoid glycoside (CIG) on the learning-memory ability and pathological changes in the brain of vascular dementia (VD) rats; To discuss relevant mechanism of action. Methods SD rats were randomly divided into sham-operation group, model group, positive medicine group, CIG groups low-, medium- and high-dose groups. The animal model of VD was replicated by permanent bilateral common carotid artery occlusion (2VO) in rats. Drugs were intragastrically administered 6 h after surgery and then once a day for 3 months. Morris water maze test was used to detect spatial learning-memory ability, and recognition memory was measured by the object recognition test. Immunohistochemical staining was used to detect the neuronal survival and the expression of choline acetyltransferase (ChAT) in the brain. Results Three months after permanent 2VO operation, the model rats showed a longer escape latency in Morris water maze, a lower discrimination index in the object recognition test, and a decrease in NeuN positive neuronal survival and ChAT expression in the hippocampus and cerebral cortex. Compared with the model group, intragastric administration of CIG for 3 months shortened the escape latency in Morris water maze, elevated the discrimination index in the object recognition test, and increased the NeuN positive neuronal survival in the hippocampus and cerebral cortex and ChAT expression of 2VO rats. Conclusion CIG can improve the cognitive impairment of VD model rats through protecting neurons and promoting ChAT expression.

Angiogenesis promotion in peri-infarction region by ChAT-positive neurons in subventricular zone after ischemic stroke / 实用医学杂志

Objective To observe the activities of ChAT + neurons in subventricular zone (SVZ) after ischemic stroke and their effects on angiogenesis in peri-infarction region and related signaling pathways. Methods C57BL/6 mice were randomly assigned into sham group,middle cerebral artery occlusion (MCAO) group and atropine group. Ischemic models were made by permanent coagulation of the distal middle cerebral artery. The expression of ChAT,AChE in SVZ and VEGF,VEGFR2,pERK in peripheral regions of ischemic injury was evaluated by Western blotting and immunofluorescence. 5-bromodeoxyuridine(BrdU)/CD31 double-labeled cells were also tested by immunofluorescence. Results At 14 d after the surgery,the ratio of ChAT/AChE in SVZ increased after stroke(P < 0.05). Compared with those in Sham group,the levels of VEGF,VEGFR2 and pERK were higher in MCAO group(P<0.05)and VEGFR2-positive and BrdU/CD31-positive cells increased significantly. However,lower expression of VEGF,VEGFR2 and pERK and less VEGFR2-positive and BrdU/CD31-positive cells were found in atropine group when compared with that in MCAO group. Conclusions The activities of ChAT +neurons in SVZ are enhanced after ischemic injury and they can promote angiogenesis in peripheral region of ischemic injury via upregulating VEGF-VEGFR2 signaling pathway and improving the brain function restoration.

Effect of Electroacupuncture at Points Xiaohai and Xiajuxu on ChAT andα 7 nAchR in a Rat Model of DU / 上海针灸杂志

Objective To investigate the effect of electroacupuncture at points Xiaohai and Xiajuxu on the expressions of tumour necrosis factor-α (TNF-α) and choline acetyltransferase (ChAT) in serum and nicotinic acetylcholine receptorα 7 (α 7 nAchR) in duodenal tissues in a rat model of duodenal ulcer (DU) and preliminarily explore the relative specificity of He-Sea point in “treating visceral diseases with He-Sea point”.Methods Forty healthy SD rats were randomized into blank (A), model (B), Xiaohai (C) and Xiajuxu (D) groups, 10 rats each. A rat model of DU was made by subcutaneous injection of 10% cysteamine hydrochloride at the right buttock. After successful model making, group C was given electroacupuncture at point Xiaohai and group D, at point Xiajuxu. Duodenal tissue ulcer was macroscopically observed and scored in every group of rats. Rat serum expression of TNF-α was determined by double antibody sandwich enzyme-linked immunosorbent assay (ELISA); rat serum expression of ChAT, by ultraviolet spectrophotometry & colorimetry; rat duodenal expression ofα7 nAchR, by Western blot.Results After model making, the duodenal ulcer score was significantly higher in groups B, C and D than in group A (allP0.05) and was significantly higher in group D than in group B (P<0.01) or C (P<0.05).α7 nAchR expression was significantly higher in groups C and D than in group B (bothP<0.01). There was a positive correlation between ChAT andα7 nAchR expressions in every group (r=0.444,P=0.007).Conclusions Electroacupuncture at both points Xiaohai and Xiajuxu can reduce the duodenal ulcer score and serum TNF-α expression and increase serum ChAT and duodenalα7 nAchR expressions in DU rats. The results show that the therapeutic effect of electroacupuncture on duodenal ulcer may be produced by regulating TNF-α. Its mechanism may be activating cholinergic anti-inflammatory pathway to produce an anti-inflammatory effect. The effect being better in group D than in group C suggests that Xiajuxu has the relative specificity.

The Effect of Choline Acetyltransferase Genotype on Donepezil Treatment Response in Patients with Alzheimer's Disease

OBJECTIVE: We examined the difference in responses to donepezil between carriers and non-carriers of the A allele at the +4 position of the choline acetyltransferase (ChAT) gene in Koreans. METHODS: Patients who met the criteria for probable Alzheimer's disease (AD) (n=199) were recruited. Among these, 145 completed the 12-week follow-up evaluation and 135 completed the 26-week scheduled course. Differences and changes in the Korean version of the mini-mental state examination (MMSE-KC) score, Korean version of the Consortium to Establish a Registry for Alzheimer's Disease Neuropsychological Assessment Battery (CERAD-K[N]) wordlist subtest score (WSS), CERAD-K(N) total score (TS), and the Korean version of geriatric depression scale (GDS-K) score between baseline and 12 weeks or 26 weeks were assessed by the Student's t-test. RESULTS: At 12 weeks, the changes in the MMSE-KC score, CERAD-K(N) WSS, and CERAD-K(N) TS from baseline were not significant between ChAT A allele carriers and non-carriers; however, at 26 weeks, these changes were significantly larger in ChAT A allele carriers than in non-carriers (p=0.02 for MMSE-KC and p=0.03 for CERAD-K(N) WSS respectively). CONCLUSION: Our findings in this study suggested that presence of the A allele at the +4 position of ChAT might positively influence the treatment effect of donepezil in the early stages of AD in Koreans.

Effects of chronic cerebral ischemia on hippocampal microvascular architecture and ChAT expression in rats / 吉林大学学报(医学版)

Objective To investigate the damage of hippocampal neurons induced by chronic cerebral ischemia in the rats,and to clarify its mechanism.Methods 90 healthy adult male SD rats were randomly divided into sham operation group (n=30 ) and experimental group (n=60 ). The chronic cerebral ischemia rat models were established by permanently ligating the common carotid arteries on both sides of the rats in experimental group.The rats in sham operation group were established by incising the cervical median,only the common carotid arteries on both sides were separated without ligating. The rats in sham operation group and experimental group were respectively sacrificed at the 7th,14th and 21st day after operation.At each time point 10 rats in sham operation group and 20 rats in experimental group were selected and sacrificed.Immunohistochemistry staining was used to observe the dynamic changes of the expression levels of choline acetyltransferase(ChAT)in hippocampus neurons, and TA-FE method was used to observe the dynamic changes of hippocampal microvascualr architecture. MiVnt image analytical system was used to quantitatively analyze the immunohistochemistry result,the microvessel density (MVD)and micorvessel area density (MVA)of horizontal part of hippocampus in the rats. Results Compared with sham operation group,the ChAT expression levels in hippocampus neurons of the rats in experimental group at different time points were significantly decreased(P<0.05);and with the prolongation of time,the ChAT expression levels were gradually decreased;the ChAT expression level in 14-day experimental group was significantly lower than that in 7-day experimental group (P<0.05 );the ChAT expression level in 21-day experimental group was significantly lower than those in 7-day and 14-day experimental groups(P<0.05).The MVD and MVA of hippocampus of the rats in experimental group at different time points were obviously decreased compared with sham operation group(P<0.05);the MVA was gradually decreased with the prolongation of time, and the MVA of hippocampus of the rats in 21-day and 14-day experimental groups were obviously decreased compared with 7-day experimental group(P<0.05);the MVD was gradually decreased with the prolongation of time,the MVD of hippocampus of the rats in 21-day and 14-day experimental groups was obviously decreased compared with 7-day experimental group(P<0.05);the MVD of hippocampus of the rats in 21-day experimental group was obviously decreased compared with 14-day experimental group(P<0.05).Conclusion Chronic cerebral ischemia can lead to the progressive decrease of the ChAT expression level,MVD and MVA of hippocampus of the rats to aggravate gradually the learning and memory dysfunction, which may be one of the reasons of vascular dementia.

Effects of exercise training on cognitive function,ChAT and AchE activity in SHR/SP rat vascular dementia model / 重庆医学

Objective To investigate the effects of the exercise training on the cognitive function ,choline acetyltransferase (ChAT) activity and acetylcholinesterase (AchE) activity in stroke prone spontaneously hypertensive rat (SHR/SP) vascular de-mentia model .Methods 30 male SHR/SP rats were randomly divided into sham operation group ,model group and exercise group (n=10) .The VD model was established by the fractional ligation of bilateral carotid artery (2-VO) .The sham operation group and the model group were given the normal feeding without intervention after operation ;the exercise group adopted the treadmill exer-cise(DSPT-1) for 8 weeks .After the exercise ,the Morris maze test was conducted for evaluating the cognitive function in each group .The rats were finally killed for detecting the ChAT activity and AchE activity of hippocampus .Results In the positioning navigation training ,the latency period of the sham operation group was significantly short than that of the exercise group and the model group ,but the latency period of the exercise group was obviously short than that of rats in the model group (P<0 .05);in the spatial exploration test ,the rats in the sham operation group had more frequency of crossing platform than the other two groups ,the exercise group had more frequency of crossing platform in platform quadrant than the model group (P<0 .05);the exercise training could increase the ChAT activity and lower the AchE activity of hippocampus .Conclusion The exercise training can improve the function of hippocampal cholinergic system in SHR/SP and then increase the cognitive ability .

Effects of melatonin on choline acetyltransferase in rat hippocampus after isoflurane anesthesia / 中华麻醉学杂志

Objective To investigate the effects of melatonin on choline acetyltransferase (ChAT) in the hippocampus of rats after isoflurane anesthesia.Methods Sixty male SD rats weighing 390-440 g were randomized into five groups (n =12 each):control group (group C),1％ isoflurane group (group Ⅰ),1％ isoflurane + melatonin group (group IM),2％ isoflurane group (group J) and 2％ isoflurane + melatonin group (group JM).Rats in groups IM and JM received intraperitoneal injection of melatonin (10 mg/kg) for 7 days,and rats in other groups received normal saline.On the 7th day of injection,rats in groups Ⅰ and IM inhaled 1％ isoflurane for 4 hours,and rats in groups J and JM inhaled 2％ isoflurane for 4 hours.One day after anesthesia,all the rats began Morris water maze to assess the learning and memory ability,which was made for continuous 5 days.At the end of probe test,6 rats in each group were randomly selected,blood samples were collected to detect plasma melatonin level,and the hippocampi were removed to evaluate the expression and activity of ChAT.The other rats were sacrificed to perform immunofluorescence to detect ChAT in hippocampal CA1 region and dentate gyrus.Results The plasma melatonin level,and the expression and activity of ChAT were significantly lower in group Ⅰ than in group C (P ＜ 0.01).The escape latency was significantly longer,the probe time was significantly shorter,and the plasma melatonin level and the expression and activity of ChAT were significantly lower in group J than in group C (P ＜ 0.05 or 0.01).The escape latency was significantly shorter,the probe time was significantly longer,and the plasma melatonin level and the expression and activity of ChAT were significantly higher in group IM than in group Ⅰ (P ＜ 0.05 or 0.01).The escape latency was significantly shorter,and the plasma melatonin level and the ChAT activity were significantly higher in group JM than in group J (P＜ 0.05 or 0.01).Conclusion Melatonin can attenuate isoflurane-induced ChAT inhibition and thus improve the cognitive function of rats after isoflurane anesthesia.

Study on Pathology and Neurotransmitters of Esophageal Sphincter after Radiofrequency Thermocoagulation in Dogs / 天津医药

Objective To investigate the effects of radiofrequency thermocoagulation (RFT) on pathological features and the expressions of choline acetyltransferase (ChAT), vasoactive intestinal peptide (VIP) and nitric oxide synthase (NOS) at lower esophageal sphincter (LES) in family dogs. Methods A total of 15 dogs were randomly divided into three groups. Sham group underwent gastroscopy and was fed for 3 months (n=5). Dogs were given RFT and were fed for 24 h after RFT (n=5, RFT+24 h group). Dogs were given RFT and were fed for 3 months after RFT (n=5, RFT+3m group). The pathological changes of LES were observed after HE staining in three groups. The expressions of ChAT, VIP and NOS were detected by immunohistochemical method in three groups. Results Results of HE staining showed nearly the same tissues in Sham group and control group. There were active inflammatory reaction and structural damage in RFT+24 h group. The chronic in-flammatory reaction and structural remodeling were found in RFT+3m group. Immunohistochemistry showed that ChAT was significantly increased in RFT+3m group compare than that of Sham group. Values of VIP and NOS were significantly de-creased in RFT+3m group compare than that of Sham group (P<0.01). Conclusion The thickness and increased pressure of LES were found after RFT,which also caused changes in neurotransmitters of local tissues in dogs.

Study of the activities of ChAT and CHE in patients with non-dementia vascular cognitive impairment / 重庆医学

Objective To study of the activities of choline acetyltransferase (ChAT ) and cholinesterase(CHE) in non-dementia vascular cognitive impairment (VCIND)patients ,and explore its mechanism .Methods Sixty-one patients with VCIND were select-ed as the research group (which accord with Rockwood diagnostic criteria for cognitive dysfunction but not meet the NINDS-ARI-IEN diagnostic criteria for vascular dementia)and 75 healthy people in the same period as the control group .Venous blood was col-lected in the early morning and serum was separated in both of the two groups .The activities of ChAT and CHE were detected by spectrophotometric method .Results The activity of ChAT in the research group(120 .94 ± 23 .93) U/mL was increased significant-ly than the control group(64 .88 ± 12 .23) U/mL(P0 .05) .Conclusion The increased activity of ChAT in VCIND patient and enhance the synthesis of acetylcholine , which may be a compensatory mechanism of VCIND .

Effect of Heixiaoyao Powder on Mice Model of Senile Dementia Induced by Scopolamine / 中国中医药信息杂志

Objective To explore the mechanisms and the significance of compatibility of Heixiaoyao Powder by observing the effect of Heixiaoyao Powder on mice model of senile dementia induced by scopolamine. Methods Kunming mice were randomly divided into five groups including control group, model group, Heixiaoyao Powder group, Xiaoyao powder group and streamlined formula group, with 10 mice of each group. Scopolamine hydrobromide intraperitoneal injection was adopted to make memory impairment in mice model, and followed by drugs treatment with corresponding decoction of the crude drug 2.314 g /(kg·d). Control group and model group were given normal saline orally once a day, all groups were treated for 10 days continuously. After 6 and 8 days of the administration, water maze test, stair jumping test and darkness avoidance test were conducted to test behavior proficiency, and the activity of acetylcholinesterase (AchE) activity and choline acetyltransferase (ChAT) in brain tissue were tested also. Results Compared with model group, Heixiaoyao Powder group, Xiaoyao Powder group and streamlined formula group could significantly reduce the error number and shorten latency in the water maze test, decrease the time of electrical shocks and prolong the incubation period of time in both stair jumping test and darkness avoidance test, and could increase ChAT activity and lower AchE activity in brain tissue of mice (P<0.01 or P<0.05), especially the effects of Heixiaoyao Powder group. Conclusions Heixiaoyao Powder and its analogous formula have the function of preventing and treating senile dementia.