EFFECTS OF MOXIBUSTION ON CEREBRAL ACETYLCHOLINE CONTENT AND CHOLINE ACETYL TRANSFERASE ACTIVITY IN THE AGED RATS

Objective To observe the effect of moxibustion of Baihui（百会GV20） and Shenshu（肾俞 BL 23） on acetylcholine （AOh） content and choline acetyl transferase （CHAT） activity in the brain of aging rats so as to investigate its underlying mechanism in delaying brain aging in the rat. Methods Thirty Wistar rats including 10 young rats （young group） and 20 aged rats that were divided into aging group and moxibustion group evenly, were used in this study. For rats of moxibustion group, moxibustion was applied to Baihui （百会GV20） and Shenshu（肾俞 BL23） for 10 min, once a day, with 5 days being a course of treatment, 8 courses altogether. At the end of the experiments, the rats anesthetized with 6 % chloral hydrate were decapitated for taking brain tissue, then AOh content, CHAT and acetylcholinesterase （ACHE） activity were detected by using high performance liquid chromatography （HPLC）. Results Compared with young rat group, AOh contents of basal ganglia, hippocampus and cerebral cortex tissues in aging group and moxibustion group all decreased significantly （P〈 0.05, 0.01 ）, while compared with aging group, ACh contents in the 3 cerebral regions in moxibustion group all increased considerably （P〈0.01）. In comparison with young group, both CHAT activity and AChE activity of the brain tissue in aging group and moxibustion group lowered significantly （ P 〈 0.05, 0.01 ） ; comparison between aging group and moxibustion group showed that CHAT activity of the later group increased evidently （P〈 0.05） while AChE activity in moxibustion group decreased considerably （P〈0.01）, displaying that moxibustion could potentiate CHAT activity and further lower AChE activity of the brain in aged rats. Conclusion Moxibustion of Baihui（百会GV20） and Shenshu（肾俞 BL23） has effects in raising AOh contents and lowering CHAT and AChE activity, which may contribute to its efficacies in repairing the injured central cholinergic nerve system and delaying the aging process in the aged rats.

PROLIFERATION AND DIFFERENTIATION OF NEURAL STEMCELLS IN ADULT RATS AFTER CEREBRAL INFARCTION

Objective To investigate proliferation and differentiation of neural stem cells in adult rats after cerebral infarction. Methods Models of cerebral infarction in rats were made and the time-course expression of bromodeoxyuridine(BrdU), Musashi1, glial fibrillary acidic protein (GFAP), and neuronal nuclear antigen (NeuN) were determined by immunohistochemistry and immunofluorescence staining. BrdU and Musashi1 were used to mark dividing neural stem cells. GFAP and NeuN were used to mark differentiating neural stem cells. Results Compared with controls, the number of BrdU-labeled and BrdU-labeled with Musashi1-positive cells incre-ased strikingly 1 day after cerebral infarction; approximately 6 fold with a peak 7 days later; markedly decreased 14 days later, but was still elevated compared with that of controls; decling to the control level 28 days later. The number of BrdU-labeled with GFAP-positive cells nearly remained unchanged in the hippocampus after cerebral infarction. The nu-mber of BrdU-labeled with NeuN-positive cells increased strikingly 14 days after cerebral infarction, reached maximum peak in the hippocampus 28 days after cerebral infarction in rats. Conclusion Cerebral infarction stimulate proliferation of inherent neural stem cells and most proliferated neural stem cells differentiate into neurons.

Effect of ginkgo biloba extract on livers in aged rats

AIM: To investigate the protective effect of ginkgo biloba extract (GBE) on livers of aged rats and the associated mechanisms.METHODS: Two-mo- and 20-mo-old rats were treated with GBE/saline for 3 mo. Liver tissue samples from 5-moold rats treated with saline (group Y) and 23-mo-old rats treated with GBE (group E) or saline (group N) were used for histopathological examinations (hematoxylin-eosin and Masson staining, Lipofuscin staining-Schmorl staining) and determination of expression of tissue inhibitor-1 of metalloproteinase (TIMP-1) and the level of malondialdehyde (MDA), glutathione peroxidase (GPx) and superoxide dismutase (SOD). Blood samples were collected for determination of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL) and albumin.RESULTS: Microscopic studies with Masson staining revealed mild liver fibrosis in aged rats (group N), while the livers of aged rats receiving GBE (group E) showed amelioration in fibrosis (2.2±0.1 vs 2.8±0.1, P＜0.01) and deposition of lipofuscin (33.7±5.3 vs 62.8±5.7, P＜0.01).The expression of TIMP-1 and the level of liver MDA (1.0±0.1 vs 1.2±0.2, P＜0.05) also decreased but the activity of GPx (97.1±15.3 vs 61.8±14.5, P＜0.01) increased in group E. Compared with group Y, the level of liver MDA (0.8±0.1 vs 1.2±0.2, P＜0.01), lipofuscin (32.4±6.0 vs 62.8±5.7, P＜0.01) and TIMP-1 expression were increased,while the activity of GPx (103.2±17.6 vs61.8±14.5, P＜0.01)and SOD (16.7±4.4 vs 11.8±3.9, P＜0.05) was decreased in group N. There was no difference in liver function among these three groups.CONCLUSION: GBE has protective effects on aging liver.The possible mechanisms might be its antioxidant activity and inhibition of TIMP-1 expression.

Effect of different intensities of physical activity on cardiometabolic markers and vascular and cardiac function in adult rats fed with a high-fat high-carbohydrate diet

Background: Physical activity(PA) and diet are 2 lifestyle factors that affect cardiometabolic risk. However, data on how a high-fat highcarbohydrate(HFHC) diet influences the effect of different intensities of PA on cardiometabolic health and cardiovascular function in a controlled setting are yet to be fully established. This study investigated the effect of sedentary behavior, light-intensity training(LIT), and high-intensity interval training(HIIT) on cardiometabolic markers and vascular and cardiac function in HFHC-fed adult rats.Methods: Twelve-week-old Wistar rats were randomly allocated to 4 groups(12 rats/group): control(CTL), sedentary(SED), LIT, and HIIT.Biometric indices, glucose and lipid control, inflammatory and oxidative stress markers, vascular reactivity, and cardiac electrophysiology of the experimental groups were examined after 12 weeks of HFHC-diet feeding and PA interventions.Results: The SED group had slower cardiac conduction(p = 0.0426) and greater thoracic aortic contractile responses(p < 0.05) compared with the CTL group. The LIT group showed improved cardiac conduction compared with the SED group(p = 0.0003), and the HIIT group showed decreased mesenteric artery contractile responses compared with all other groups and improved endothelium-dependent mesenteric artery relaxation compared with the LIT group(both p < 0.05). The LIT and HIIT groups had lower visceral(p = 0.0057 for LIT, p = 0.0120 for HIIT)and epididymal fat(p < 0.0001 for LIT, p = 0.0002 for HIIT) compared with the CTL group.Conclusion: LIT induced positive adaptations on fat accumulation and cardiac conduction, and HIIT induced a positive effect on fat accumulation,mesenteric artery contraction, and endothelium-dependent relaxation. No other differences were observed between groups. These findings suggest that few positive health effects can be achieved through LIT and HIIT when consuming a chronic and sustained HFHC diet.

Comparative study of histopathology changes between the PS1/APP double transgenic mouse model and Aβ_(1-40)-injected rat model of Alzheimer disease

Objective To identify the genetype of the PS1/APP double transgenie mouse model, then to analyse the histopathological changes in the brain and compare the differences between the transgenie mice models and Aβ1-40-injeeted rats models of Alzheimer disease. Methods The modified congo red staining, Nissl＇s staining and immunohistology staining was used to observe the Aβ deposits, activation of astrocyte respectively. Results ①The PS1/APP transgenic mouse extensively displayed Aβ deposits in the cortex and hippocampal structures, and GFAP positive cells were aggregated in mass and surrounded the congo red-positive plaque. ②The Aβ1-40-intrahippocmnpal-injeeted rat model showed the Aβ plaque deposits in the dentate gyrus of the hippocampus, with the astrocyte surrounded. The neurons loss was significant in the injection point and pin hole of injection with Nissl＇s staining methods. GFAP-positive cells increased significantly compared with the uninjected lateral of the hippocampus. Conclusion Although Aβ1-40-injected rat models could simulate some characteristic pathological features of human Alzheimer diseases, Aβ deposits and neurons loss in partial hippocampal, it would not simulate the progressive degenenration in the brain of AD. The double transgenie PS1/APP mice could simulate the specific pathogenesis and progressive changes of AD, mainly is Aβ deposits and the spongiocyte response , while no neurons loss were observed in this model.

Responses of the Adult Rat Glucose Metabolism to Early Life Feeding, Caloric Restriction and Refeeding

Early life overfeeding in the rat can be experimentally induced by reducing litter size. This investigation assessed the consequences of this manipulation on glucose metabolism in vivo and in isolated hepatocytes in 150-day old rats. Additionally, after body growth, the effects of caloric restriction and refeeding were tested. Adult rats from control （G9） and reduced litters （G3L） did not differ in body and fat weights, glucose tolerance or insulin resistance （insulin-induced hypoglycemia）, or hepatocyte glucose release under basal or gluconeogenic conditions. Caloric restriction （G3R） reduced body and fat weights, decreased glucose decay after insulin injection and decreased hepatocyte gluconeogenic glucose release. Refeeding after caloric restriction reversed these parameters to those of the freely-fed groups （G9 and G3L）. Taken together, these results suggest that the liver glucose metabolism is not programmed by lactational overfeeding, but rather is responsive to the current nutritional condition of the animal.

Clinical Observation and Experimental Study on Compound Hypoglycemic Decoction for Hyperglycemia

Purpose: To observe the clinical efficacy of Compound Hypoglycemic Decoction (CHD) and its effect on serum total cholesterol in model mice. Method: The paired t test was used to analyze the data recorded before and after administration of drugs for hyperglycemia induced by intraperitoneal injection of 75% egg yolk emulsion in experimental mice. CHD and Fenofibrate were administered as prevention measures. Results: The total effective rate of serum total cholesterol (TC) decrease in 60 cases of hyperglycemia was 86.66% and that of serum total triglyceride (TG) decrease was 81.81%. The total effective rate of high-density lipoprotein-cholesterol (HDL-C) increase was 75%. The decrease in TC and TG, and the increase of HDL-C after treatment by in-group comparison were all significant (P<0.05). 21.9% and 22.2% decrease in the total cholesterol was respectively found in the CHD and Fenofibrate groups (both P<0.05), with no significant difference. Conclusion: The hypoglycemic action of Compound Hypoglycemic Decoction was remarkable in clinical practice, and it is very effective in preventing hyperglycemia in experimental mice.

Protective Effect of Rhubarb on Endotoxin-Induced Acute Lung Injury

To approach the mechanism of lipopolysaccharide (LPS) in causing acute lung injury (ALI) and the protective effect of rhubarb and dexamethasone, lung specimens were examined with macroscopy, microscopy, electron microscopy and the biological markers of ALI including lung wet/dry weight, the rate of neutrophils and protein content in the pulmonary alveolar lavage fluid, pulmonary capillary permeability and pulmonary alveolar permeability index were observed. The mechanism of the ALI is mainly due to direct injury of alveolar epithelium and pulmonary vascular endothelium. Rhubarb and dexamethasone could significantly reduce the edema of the lung tissue, decrease the red blood cell exudation, neutrophil infiltration and plasma protein exudation in the alveoli and all the biological markers in comparison with the ALI model rats, indicating they have protective action on vascular endothelium and alveolar epithelium.

Actions of NO and INOS on Endotoxin Induced rat Acute Lung Injury and Effect of Rhubarb on Them

This study is to explore the actions of nitric oxide (NO) and inducible nitric oxide synthase (iNOS) on endotoxin (lipopolysaccharide, LPS) induced rat acute lung injury (ALI) and effect of Rhubarb on them. LPS was injected into the sublingual vein of male Wistar rats to prepare ALI animal models. The rats were divided into 4 groups: LPS, control, Rhubarb, and dexamethasone. Macroscopic and histopathological examinations of the lung specimens were performed and the biological indexes of lung, including wet weight/dry weight, the rate of neutrophils and protein content in the pulmonary alveolar lavage fluid, pulmonary vascular permeability and pulmonary alveolar permeability were observed. In the mean time, the contents of serum NO and the activities of lung tissue homogenate iNOS were measured. The results showed that in the LPS group, the injury and celluar infiltration in the pulmonary stroma and alveoli were more prominent than that in the control group. Lung wet weight/dry weight, the rate of neutrophils, protein content, pulmonary alveolar permeability, pulmonary vascular permeability were significantly increased (P<0.01); NO and iNOS were also markedly elevated (P<0.01). In the groups of dexamethasone and Rhubarb, the histopathological changes were significantly milder, and all the above biological indexes of lung injury and the contents of NO and the activities of iNOS were correspondingly decreased (P<0.05). The above data demonstrate that NO and the activities of iNOS play an important role in the onset of ALI; dexamethasone and Rhubarb interfering treatment can ameliorate lung injury and decrease the concentrations of NO and iNOS, showing that through inhibiting the levels of NO and the activities of iNOS, these 2 agents exert protective effect on ALI induced LPS.

Application of the sodium hyaluronate-CNTF scaffolds in repairing adult rat spinal cord injury and facilitating neural network formation

The present study aimed to explore the potential of the sodium hyaluronate-CNTF (ciliary neurotrophic factor) scaffold in activating endogenous neurogenesis and facilitating neural network re-formation after the adult rat spinal cord injury (SCI). After completely cutting and removing a 5-mm adult rat T8 segment, a sodium hyaluronate-CNTF scaffold was implanted into the lesion area. Dil tracing and immunofluorescence staining were used to observe the proliferation, differentiation and integration of neural stem cells (NSCs) after SCI. A planar multielectrode dish system (MED64) was used to test the electrophysiological characteristics of the regenerated neural network in the lesioned area. Electrophysiology and behavior evaluation were used to evaluate functional recovery of paraplegic rat hindlimbs. The Dil tracing and immunofluorescence results suggest that the sodium hyaluronate-CNTF scaffold could activate the NSCs originating from the spinal cord ependymal, and facilitate their migration to the lesion area and differentiation into mature neurons, which were capable of forming synaptic contact and receiving glutamatergic excitatory synaptic input. The MED64 results suggest that functional synapsis could be established among regenerated neurons as well as between regenerated neurons and the host tissue, which has been evidenced to be glutamatergic excitatory synapsis. The electrophysiology and behavior evaluation results indicate that the paraplegic rats’ sensory and motor functions were recovered in some degree. Collectively, this study may shed light on paraplegia treatment in clinics.

Age-dependent expression of forkhead box O proteins in the duodenum of rats

The O subfamily of forkhead box(FoxO) proteins is the downstream effector of the insulin-like growth factor-1/phosphoinositide 3-kinase/protein kinase B(IGF-1/PI3K/PKB) signal pathway.The objective of the present study was to examine the expressions of three members of FoxO proteins,FoxO1,FoxO3a,and FoxO4 in the duodenum of Sprague-Dawley rats at different ages.The result demonstrated that the expression of FoxO4 in rat duodenum showed an age-dependent manner.At Day 21,there were no detectable localization and expression of FoxO4 in the duodenum,while,at Months 2 and 6,localization and expression of FoxO4 were distinct.In addition,FoxO4 staining was primarily concentrated in the cell nuclei of the lamina propria around the intestinal gland of the duodenum in 2-month-old rats,but was not detectable in the same area in 6-month-old rats.Our results showed also that although FoxO3a existed in the cytoplasm of the lamina propria at a low level at the 2-and 6-month marks,it was still not detectable at Day 21.Besides,FoxO1 was not detectable in all parts and stages.Taken together,our findings suggested that the cell-specific and age-dependent expressional patterns of FoxO4 and FoxO3a proteins in the duodenum play some roles in the development and growth performance of the rat duodenum.