Effects of Chinese traditional medicine-selected recipe Q0409 on ability of learning and memory in SAM-P/8 mice / 中国病理生理杂志

AIM:To investigate the effects of Chinese traditional medicine-selected recipe Q0409 on the ability of learning and memory in SAM-P/8 mice. METHODS:Total 91 mice (4-month-old SAM-P/8 mice, SAM-R/1 mice and Kunming mice) were used in the study, in which the male and female animals were labeled separately. According to the performance of Morris water maze test, the mice were divided into 5 groups randomly. The mice were fed with different drugs or distilled water for 60 d (from 4 months to 6 months). The mice were fed with the drugs from 61 d to 65 d, and 1 h later each time, the Morris water maze test was carried out. After this Morris test were finished at 65 d, the mice were killed immediately and their hippocampal tissues were isolated. Half of the hippocampal tissues were added with precooled normal saline and made into 10% (g/mL) homogenate for detecting the protein content and acetyl cholinesterase (AChE) activity. The other half was fixed with 4% paraformaldehyde and embedded with paraffin for immunohistochemical staining of amyloid β-protein (Aβ). RESULTS:Compared with model group, the results of navigation training and spatial probe training in Morris water maze test were significantly improved (P<0.05), and the activity of AChE in the hippocampal ho-mogenate was significantly decreased (P<0.05) in Q0409 treatment group. No difference in Q0409 group was observed compared with control group and positive drug (huperzine A) group. Immunohistochemical staining showed no typical "se-nile plaques" in the male mice of Q0409 group, while there was shallower and smaller brown staining in the hippocampus of the female mice of Q0409 group. The positive area of Aβ deposition decreased in the CA1 area of hippocampal tissues in Q0409 group. These results were similar to those in positive drug group. CONCLUSION:Q0409 improves the ability of learning and memory in SAM-P/8 mice, which is related to the inhibition of AChE activity and the reduction of Aβ protein deposition in the hippocampus. The effects is similar to those of huperzine A.

Attenuation of Neuronal Damage by Gymnemic acid in Experimentally Induced Cerebral Ischemia in Rats.

Gymnemic acid is obtained from the natural resource and has got antioxidant property, was investigated for the possible neuroprotective effect in experimentally induced cerebral ischemic rats. Carotid arteries clamped with the help of aneurysm clips to produce cerebral ischemia, the clips were removed from the arteries to allow the reflow of the blood through carotid arteries. It was observed that lipid peroxidation was increased significantly after bilateral common carotid artery occlusion. Brain endogenous antioxidant GSH and total protein levels were low. Administration of gymnemic acid in a dose of 250 mg/kg and 500 mg/kg b.w orally showed promising neuroprotective effect by reducing cerebral infarct size as well as improved all antioxidant levels showing activity against oxidative stress.

Research progress on chemical constituents and pharmacological effects of alkaloids from plants of Lycoris Herb / 中草药

Recently, studies on Lycoris Herb. type alkaloids received the attention of scholars home and abroad. Lycoris Herb. type alkaloids can be devided into seven types according to their molecular structure, including lycorine type, crinine type, galanthamine type, tazettine type, narciclasine type, lycorenine type, homolycorine type, and montanine type. Researches have shown that Lycoris Herb. type alkaloids possess multiple phamocology activities, such as strong antitumor activity on human breast cancer cell (MCF-7), human leukemia cell (HL-60); and strong inhibitory effect of flu virus, measles virus, polio virus, and SARS virus; Lycorine type and crinine type alkaloids have antimalaria effect; Besides, lycorine type has strong anti-acetylcholinesterase effect. In a word, lycorine type and lycoris type alkaloids carry multiple pharmacological effect and belong to promising substances.

Kinetics for the Inhibition of Serum Acetylthiocholin Esterase Activity by Some Prepared Phenobarbital Derivatives.

This work addresses the kinetic analysis on the interaction of some prepared Phenobarbital derivatives (A, B, C and D) with human serum acetylcholinesterase. It was found that these compound (A, B and D) does have inhibitory effects at different concentrations (10-4, 10-6, 10-8, 10-10M), and were observed to have elevated inhibition with increasing concentrations(10-10 to 10- 4M) of concentrations for both compounds A and B, elevated inhibition with decrease concentration from 10-4 to 10-10M for D. The effects of each A, B and D were reversible in nature. All of the results for C compound were neglected. Michaelis- Menten constant and maximum velocity for hydrolysis of acetylthiocholine iodide by AChE was determined in control and treated systems. Line weaver- Burk plot and their secondary replots indicated that the nature of inhibition was (competitive at concentration 10-4, non competitive at 10-10 for A), (non competitive at 10-4, competitive at 10-10 for B), (non competitive at 10-4, and uncompetitive at 10-10 for D) respectively. The value of ki was also estimated. The action mechanism of these types of compounds acting as inhibitors to the AChE is suggested.

Effect of acute exposure of triazophos on oxidative stress and histopathological alterations in liver, kidney and brain of Wistar rats.

Acute dose of organophosphorus pesticide Triazophos (O,O-diethyl O-1-phenyl-1H-1,2,4-triazol-3-yl phosphorothioate; Tz) administered orally affects oxidative stress parameters and the histo-architecture of liver, kidney and brain tissues. The results indicate a dose dependent induction of oxidative stress as evident by increased malondialdehyde level and decreased antioxidant defense including glutathione and superoxide dismutase activity in rat liver, kidney and brain. AChE activity was found significantly decreased in the Tz treated groups as compared to the vehicle control (DMSO) group. Histopathological examination of liver, kidney and brain in Tz treated rats revealed medullary congestion and hydropic degeneration of hepatocytes in liver and medullary congestion in kidney. However, no significant histopathological changes were observed in brain tissues.

In vitro Pharmacological Investigations of Using Leaf Callus Extracts of Canthium parviflorum Lam.

Aims: The Canthium parviflorum Lam. is an important medicinal plant extensively used in traditional oriental herbal medicines. It is important to screen for the novel biological activities and novel chemical constituents to further validate the use of this plant extract. This was mainly intended to screen in vitro pharmacological activities with callus extracts of Canthium parviflorum. Experiments were designed according to the standard methods and processes. Place and Duration of Study: The Pharmacology Department of Laila Research Centre in Vijayawada, Andhra Pradesh, India between September-October 2012. Methodology: Callus was induced from leaf explants of Canthium parviflorum on MS medium supplemented with 2,4-Dichlorophenoxyacetic acid and the compounds were extracted from dried callus using methanol solvent with Soxhlet apparatus. Results: The Graph pad Prism Version‑5 software was used to analyze data in the form of figures. The callus extract was potentially inhibited the 5- Lipoxygenase inhibitory enzyme at significantly less IC50 value that was comparable with the standard drug inhibition. In addition, this extract was exhibited remarkable cytotoxicity with less LC50 value. However, this callus extract was shown very low potency in inhibiting the enzymes of acetyl cholinesterase, tyrosinase, alpha-glucosidase. Conclusion: The study demonstrated that callus extract of Canthium parviflorum shown more potent inhibition of 5-LOX and also remarkable cytotoxicity to be further screened for in vivo anticancer and anti-inflammatory activity.

Effects and mechanisms of chloro-oxime derivatives on spatial learning and memory dysfunction in two dementia animal models / 中国药理学通报

Aim To explore the effects and mecha-nisms of choro-oxime derivatives on spatial learning and memory impairment in Kunming mice and SD rats induced by scopolamine and Aβ1-42 , respectively. Methods 40 Kunming mice were randomly divided into 5 groups: control group, model group, donepezil treatment group, arimoclomol treatment group and TCO-2 treatment group. There were 8 mice in each group. Mice of control group were established by intra-peritoneal injection of saline, and mice of other groups were injected with scopolamine and caused memory im-pairment. Both control group and model group were treated with solvent by intraperitoneal administration;donepezil treatment group received donepezil by intra-gastric administration; arimoclomol treatment group and TCO-2 treatment group were given the correspond-ing drugs by abdominal injection, respectively. The solvent and drugs were given at the same time every morning for 8 days. Spatial learning and memory abili-ty were tested by Morris water maze from the fifth day of the drugs administration. 40 SD rats were divided into 5 groups the same as the dementia model men-tioned above. Mice of control group were established by intracerebroventricular injection of saline, and mice of other groups were injected with insoluble Aβ1-42 to be induced of memory impairment. Solvent and drugs were also delivered as mentioned above. Morris water maze was carried out from the fifth day of the drug de-livery. After that, acetyl cholinesterase activity of hip-pocampus was tested with acetyl cholinesterase reagent kit; the content of Aβ1-42 in hippocampus was meas-ured by ELISA assay kit;the expression of phosphoryl-ated tau proteins was detected by Western Blot. Re-sults In both two dementia models, choro-oxime de-rivatives could improve the spatial learning and memory ability, shorten the escape latency and increase the times of crossing the former platform. Choro-oxime de-rivatives could also inhibit the acetyl cholinesterase ac-tivity in animal brain, decrease the concentration of Aβ1-42 and the expression of phosphorylated tau pro-teins in the dementia rats’ hippocampus. Conclusions Spatial learning and memory deficits induced by sco-polamine and Aβ1-42 could be reversed by choro-oxime derivatives. It may be concerned with enhancement of the cholinergic system functions and reduction of the levels of Aβ1-42 and phosphorylated tau proteins in the brain.

Efficacy of L-carnosine against Schistosoma mansoni antigens in liver, heart, kidney and brain of rabbits.

The present investigation was an attempt to study the enhancememt effect of L-carnosine (beta alanyl-l-histidine) on the influence of vaccination on healthy (non-infected) rabbits treated with Schistosoma mansoni egg antigens, cercariae antigens, and worms antigens as protective agents against infection. This study involved individual injection of three Schistosoma mansoni antigens: soluble egg antigen (SEA), cercarial antigen preparation (CAP) and soluble worm antigen preparation (SWAP), in three rabbit groups, respectively. Three other groups each received the same specific antigen in conjunction with the administration of L-carnosine, biochemical parameters including DNA, RNA, DNA/RNA ratio concentrations in addition ATPase, and acetyl cholinesterase activities were measured in liver, heart, kidney and brain in all groups. Elevation in most parameters was observed in the immunized groups. Carnosine treatment of rabbit groups immunized with SEA, CAP and SWAP in comparison to the non-carnosine-treated immunized groups resulted in amelioration the changes of DNA, RNA, ATPase and acetyl cholinesterase activities in most groups. L-carnosine has a beneficial effect in the amelioration of the changes in biochemical parameters as a result of S. mansoni antigen immunization.

Dose related effects of nicotine on oxidative injury in young, adult and old rats.

Nicotine affects a variety of cellular process ranging from induction of gene expression to secretion of hormones and modulation of enzymatic activities. The objective of the present study was to study the dose dependent toxicity of nicotine on the oxidative stress in young, adult and old rats which were administered 0.75, 3 and 6 mg kg-1 nicotine as nicotine hydrogen tartarate intraperitoneally for a period of seven days. No changes were observed in blood catalase (CAT) activity and level of blood reactive oxygen species (ROS) in any of the age group at the lowest dose of nicotine. However, at the highest dose (6 mg kg-1 nicotine) ROS level increased significantly from 1.17to 1.41>M ml-1 in young rats and from 1.13 to 1.40 >M ml-1 in old rats. However, no change was observed in blood ROS levels of adult rats. Administration of 3 mg kg-1 nicotine resulted in an increase in level of reduced glutathione (GSH) in rats of all the age groups. The young animals were the most sensitive as a dose of 6 mg kg-1 resulted in decline in the levels of reduced GSH to 0.89 mg ml-1 as compared to normal control (1.03 mg ml-1). The antioxidant enzymes SOD and CAT were sensitive to a dose of 6 mg kg-1 as it resulted in decline of the enzymatic activity in all age group animals. Also, administration of nicotine at a lower dose of 3 mg kg-1 inhibited SOD activity from 1.48 to 1.20 units min-1 mg-1 protein in old rats. Catalase activity showed a similar trend at a dose of 3 mg kg-1. Administration of nicotine also increased the blood lipid peroxidation levels at all three doses in young and old rats dose dependently. Nicotine exposure also increased ROS in brain at the doses of 3 and 6 mg kg-1 in all the three age groups. Brain GSH decreased significantly at high dose of nicotine (6 mg kg-1 b.wt.) in adult rats (4.27 mg g-1) and old rats (3.68 mg g-1) but in young rats level increased to 4.40 mg g-1 at the lower dose (0.75 mg kg-1 nicotine). Brain lipid peroxidation increased at all three doses of nicotine in young as well as old rats as compared to their respective normal control. The SOD activity increased significantly in young (2.88 units min-1mg-1 protein) and old rats (1.81 units min-1mg-1 protein) as compared to their respective normal at a dose of 6 mg kg-1. Interestingly, the SOD activity decreased in adult rats (2.18 units min-1mg-1 protein) as compared to its normal control. Catalase activity decreased at the dose of 3 mg kg-1 and 6 mg kg-1 nicotine in young and old rats but no effect was observed in adult rats at any of the doses. Acetylcholine esterase (AchE) activity decreased in a dose dependent manner in adult and old rats. Overall, the results of the study indicate that young and old rats are more sensitive to nicotine induced oxidative stress as compared to the adult ones.

Antiamnesic evaluation of C. phlomidis Linn. bark extract in mice / Avaliação da atividade antiamnésica da casca de C. phlomidis Linn. em camundongos

Clerodendron phlomidis Linn. (Verbenaceae) is known as Agnimantha in sanskrit. Bark of the plant is used in treating various nervous disorders. In the present study C. phlomidis was investigated for its potential as a nootropic agent in mice. The aqueous extract of the C. phlomidis (100 and 200 mg/kg, p.o.) was administered for 6 successive days to both young and aged mice. Exteroceptive behavioral models such as elevated plus maze and passive avoidance paradigm were employed to evaluate short term and long term memory respectively. Scopolamine (0.4 mg/kg, i.p.), diazepam (1 mg/kg, i.p.) were employed to induce amnesia in mice. To delineate the mechanism by which C. phlomidis exerts nootropic action, its effect on brain acetyl cholinesterase levels were determined. Piracetam (200 mg/kg, i.p.) was used as a standard nootropic agent. Pretreatment with C. phlomidis (100 and 200 mg/kg, p.o.) for 6 successive days significantly improved learning and memory in mice. It reversed the amnesia induced by scopolamine, diazepam and natural ageing. It also decreased the acetyl cholinesterase levels in the whole brain. The bark of C. phlomidis can be of enormous use in the management of treatment of cognitive disorders such as amnesia and Alzheimer's disease.

Clerodendron phlomidis Linn. (Verbenaceae) é conhecida como Agnimantha em sânscrito. A casca da planta é utilizada no tratamento de várias disfunções neurológicas. No presente estudo, C. phlomidis foi investigada pelo seu potencial como agente nootrópico em camundongos. O extrato aquoso de C. phlomidis (100 e 200 mg/kg, p.o.) foi administrado por seis dias consecutivos tanto para camundongos jovens quanto para idosos. Modelos comportamentais exteroceptivos, tais como labirinto em cruz elevada e paradigma de esquiva passiva foram empregados para avaliar memória recente e tardia, respectivamente. Escopolamina (0,4 mg/kg i.p.), diazepam (1 mg/kg i.p.) foram empregados para induzir amnésia em camundongos. A fim de delinear o mecanismo pelo qual C. phlomidis exerce ação nootrópica, determinaram-se seus efeitos nos níveis cerebrais de acetilcolinesterase. Utilizau-se piracetam (200 mg/kg i.p.) como nootrópico padrão. O pré-tratamento com C. phlomidis (100 e 200 mg/kg, p.o.) por seis dias sucessivos melhorou, significativamente, o aprendizado e a memória em camundongos. Ela reverteu a amnésia induzida por escopolamina, diazepam e pelo envelhecimento normal. Também, diminuíram-se os níveis de acetilcolinesteraseem todo o cérebro. A casca de C. phlomidis pode ser de grande uso no tratamento de disfunções cognitivas, como amnésia e doença de Alzheimer.

Diagnostic Strategy Using Barium Enema and Rectal Suction Biopsy with Acetylcholinesterase Histochemistry in Neonates Suspected of Having Hirschsprung's Disease

To investigate the diagnostic accuracy and applicability of barium enema (BE) and rectal suction biopsy with acetyl cholinesterase (AChE) histochemistry in the diagnosis of neonatal Hirschsprung's disease (HD), we retrospectively reviewed the findings of BE and AChE staining in 96 neonates with suspected HD during a 10-year period from January 1991 to December 2000. Sixty-nine cases of HD (58 males and 11 females) and 27 cases of non-HD are included in this study. In regard to BE, HD was based on definite transitional zone, suspicious HD on reversed rectosigmoid index (RSI 1). The histochemical criterion used for the diagnosis of HD was that of Chow et al (1977), i.e., the presence of many coarse discrete cholinergic nerve fibers in the muscularis mucosae and in the immediately subjacent submucosa regardless of infiltration of cholinergic nerve fibers in the lamina propria. Of 66 neonates with HD who underwent BE, transitional zone was identified in 33 cases (50%) and reversed RSI in 19 cases (21%), microcolon in 4 cases and normal finding in 10 cases (15%) while of 27 neonates with non-HD, there was normal finding in 16 cases and reversed RSI in 9 cases (41%). Thus diagnostic accuracy based on transitional zone was 64%. The positive predictive value of reversed RSI for the diagnosis of HD was 68%. Of 42 neonates with HD who underwent AChE histochemistry, there were 41 AChE-positive reactions and one AChE-negative reaction in a neonate with total colonic aganglionosis, while of 27 cases of non-HD, there were one equivocal AChE-positive reaction and 26 AChE-negative reactions. Thus AChE histochemical study showed a 97% diagnostic accuracy with a 98% sensitivity and a 96% specificity. In conclusion, we believe that BE is valuable as a first diagnostic step since about 80% of neonates with HD show significant radiologic findings such as a transitional zone or reversed RSI. AChE histochemical study was a more reliable diagnostic tool showing a 97% diagnostic accuracy, and is particularly valuable in neonates with HD who showed reversed RSI or normal RSI on BE. A diagnostic algorithm in neonates with suspicious HD was proposed.particularly valuable in neonates with HD who showed reversed RSI or normal RSI on BE. A diagnostic algorithm in neonates with suspicious HD was proposed.

The effects of rat brain ischemia on the expression of Acetyl-cholinesterase and Caspase-3 of the immune system / 中国免疫学杂志

Objective:To explore the effects of brain ischemia on the immune system and the expression of Acetyl-cholinesterase(AChE) and Caspase-3 after neuro-immunological injury.Methods:The serum IL-4,IL-10,TNF-?,IL-2 and IL-12 concentrations and AChE level in brain,thymus and spleen of the rat brain ischemia at different time spots were detected by ELISA.The active fragments of Caspase-3 P17 and P32 in brain,thymus and spleen were tested by Western blot.The expression of AChE in nerve cells was examined immunohistochemistry.The relationship between amount of AChE and concentrations of the cytokines or NK activity as well as cell apoptosis was analyzed.Results:The concentration of AChE in brain increased gradually after ischemia,the level reached peak after 12 hours,and then decreased afterward.The AChE level in thymus and spleen increased after 1 h in accordance with the ischemic brain.The NK activity,serum TNF-?,IL-2 and IL-12 levels also increased at 0.5 h,1 h and 2 h after ischemia,but the trend reversed after 3 hours.Serum IL-4 and IL-10 levels did not change significantly at 0.5 h,1 h and 2 h after ischemia,but serum IL-10 raised after 3 hours to 48 hours.Serum IL-4 level slightly increased only appearing at ischemia 12 h point.The ratio of P17/(P17+P32) became higher with ischemic time.Conclusion:Sustained rising of AChE level due to brain ischemia induces the apoptosis of cells in the brain,thymus and spleen via activation caspase-3,leading to Th1/Th2 imbalance and immune dysfunctions.

Effect of malathion and acetylcholine on the developing larvae of Philosamia ricini (Lepidoptera: Saturniidae).

Feeding of malathion induces accumulation of acetylcholine in Philosamia ricini developing larvae via inhibition of acetyl cholinesterase activity. The insecticide also causes depletion of all nutrients, loss in weight, under-development of silk glands resulting in reduced silk production, lack of oviposition and high mortality among the insects. Acetylcholine however, while fed during the same period of development, tones up their nutritional status, induces better growth, weight gain, improved silk production and oviposition and significantly lower mortality than in the control group of insects maintained on castor leaves. This improved status of insects has been attributed to choline, the insect vitamin released from acetylcholine. Acetylcholine has also been noted to protect the insects to some extent from the poisonous effect of malathion on exposure to it after a dose of acetylcholine during the preceding instar stadium.