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.     

Effect of Bushenyisui Formula on brain tissue apoptosis and Bcl-2 in beta-amyloid protein-induced Alzheimer’s disease rat models

OBJECTIVE:To investigate the effect of Bushenyisui Formula on cell apoptosis and positive B cell lymphoma(Bcl-2) in the Brain of rat models of Alzheimer’s disease(AD) induced by beta-amyloid protein(Aβ) and the mechanism underlying the effect.METHODS:Total of 40 SD rats,20 females and 20 males,were randomly assigned to 4 groups,controlled group(A),model group(B),conventional treatment group(C) and Bushenyisui Formula treatment(BYFT) group(D),10 rats in each group.Aβ 1-42 was injected into the bilateral hippocampus of the rats in group B,C and D to create the models of AD.Sham operation was performed on the rats of group A in the same way by injecting equal volume of 0.9% sodium chloride solution into their bilateral hippocampus.5 days after operation,Bushenyisui Formula was intraperitoneally administered at a dose of 450 mg/kg to the rats of group D(QD) for 20 days.Equalvolumeof0.9%sodiumchloridesolution wasintraperitoneallyinjectedintotheratsofgroupB withthesameprocedure.C suspension(20 mg/mL) was intraperitoneally injected into the rats of group B with the same procedure.The number of apoptotic cells in Brain and the positive Bcl-2 were counted.The changes of learning and memory abilities were evaluated usingY-maze.RESULTS:Right after the establishment of the models,group B,C and D compared to group A respectively,the outcomes of Y-maze were significantly different from that of group A,which suggested obvious learning and memory disorder in those groups(P<0.01).After treatment,the times of electronic shocks of group C and D were significantly less than that of group B(P<0.05),and the numbers of apoptotic cells and positive Bcl-2 were significantly different from those of group B,apoptotic sells’ number of group C and D smaller than that of group B and the number of positive Bcl-2 greater than that of group B.CONCLUSION:Bushenyisui Formula could increase the number of Bcl-2 in brain,which improved the function of nervous system pertaining to learning and memory abilities.     

Multiple Pharmacological Actions of Yiqi Huatan Decoction (益气化痰方) in A Model of Depression in Rats

Objective: To investigate the influence of Yiqi Huatan Decoction (益气化痰方, YHD) on a model of depression in rats under different pathological conditions. Methods: Thirty-two male SD rats were randomly divided into 4 groups of 8: normal, model, YHD, and maprotiline. The model group, YHD group and maprotiline group used separate feeding and rats were exposed to chronic and unpredictable stress to build the depression model. From day 2, the YHD group and maprotiline group were respectively given YHD (7 g/kg) and maprotiline (10 mg/kg) by gastrogavage once daily. The normal and model groups were given the same volume of drinking water. The medication duration were 21 days. At the end of the experiment, the serum levels of copper and zinc were determined by atomic absorption spectroscopy, plasma concentrations of adrenocorticotropic hormone (ACTH) and cortisol (COR) were detected by radioimmunoassay, and levels of norepinephrine (NE), dopamine (DA), and 5-hydroxytryptamine (5-HT) in the hypothalamus were analysed by high performance liquid chromatography-eletricochemistry. Results: Compared with the content of copper and zinc in the serum of rats in the normal group, serum copper levels in model rats were significantly increased and zinc content was significantly reduced (both P<0.05). Plasma concentrations of ACTH and COR in the model group were significantly increased compared with those in the normal group (P<0.05, P<0.01). The contents of NE, DA, and 5-HT in the hypothalamus of rats in the model group were significantly reduced compared with those of the normal group (P<0.05 or P<0.01). Compared with those in the model group, the serum copper content and plasma concentrations of ACTH and COR were significantly decreased (all P<0.05); meanwhile, serum zinc content and hypothalamic contents of NE, DA, and 5-HT were significantly increased in rats of the YHD group (all P<0.05). The same effects were also shown in the maprotiline group except for 5-HT (all P<0.05). Conclusion: The pharmacological actions of YHD for depression might be related to improving trace-element anomalies, reversing endocrine dysfunction, and modulating the disorders of monoaminergic neurotransmitters.     

Tojapride Reverses Esophageal Epithelial Inflammatory Responses on Reflux Esophagitis Model Rats

Objective: To investigate the mechanism of Tojapride, a Chinese herbal formula extract, on strengthening the barrier function of esophageal epithelium in rats with reflux esophagitis (RE). Methods: Ten out of 85 SD rats were randomly selected as the sham group (n=10), and 75 rats were developed a reflux esophagitis model (RE) by the esophageal and duodenal side-to-side anastomosis. Fifty successful modeling rats were divided into different medicated groups through a random number table including the model, low-, medium-, and high dose of Tojapride as well as omeprazole groups (n=10). Three doses of Tojapride [5.73, 11.46, 22.92 g/(kg?d)] and omeprazole [4.17 mg/(kg?d)] were administrated intragastrically twice daily for 3 weeks. And the rats in the sham and model groups were administered with 10 mL/kg distilled water. Gastric fluid was collected and the supernatant was kept to measure for volume, pH value and acidity. Esophageal tissues were isolated to monitor the morphological changes through hematoxylin-eosin (HE) staining, and esophageal epithelial ultrastructure was observed by transmission electron microscopy. The expressions of nuclear factor kappa-light-chain-enhancer of activated B cells p65 (NF-κ Bp65), κ B kinase beta (IKKβ ), occludin, and zonula occludens-1 (ZO-1) in the esophageal tissues were measured by immunohistochemistry and Western blot, respectively. Results: The gastric pH value in the model group was significantly lower than the sham group (P<0.05). Compared with the model group, gastric pH value in the omeprazole and medium-dose of Tojapride groups were significantly higher (P<0.05). A large area of ulceration was found on the esophageal mucosa from the model rats, while varying degrees of congestion and partially visible erosion was observed in the remaining groups. Remarkable increase in cell gap width and decrease in desmosome count was seen in RE rats and the effect was reversed by Tojapride treatment. Compared with the sham group, the IKKβ levels were significantly higher in the model group (P<0.05). However, the IKKβ levels were down-regulated after treatment by all doses of Tojapride (P<0.01 or P<0.05). The occluding and ZO-1 levels decreased in the model group compared with the sham group (P<0.01 or P<0.05), while both indices were significantly up-regulated in the Tojapride-treated groups (P<0.01 or P<0.05). Conclusions: Tojapride could improve the pathological conditions of esophageal epithelium in RE rats. The underlying mechanisms may involve in down-regulating the IKKβ expression and elevating ZO-1 and occludin expression, thereby alleviating the inflammation of the esophagus and strengthening the barrier function of the esophageal epithelium.     

温化蠲痹方对于胶原蛋白性关节炎大鼠滑膜细胞增殖和凋亡的作用

Objective: To study the effect of Wenhua Juanbi Recipe (温化蠲痹方, WJR) on proliferation and apoptosis of synoviocytes in rats with collagen-inducing arthritis (CIA). Methods: A CIA model was induced by intradermal injection of bovine collagen type Ⅱ emulsion at the base of rat tails. Thirty modeled healthy Wistar rats were randomly assigned to one of three groups (10 per group): the model group, the methotrexate (MTX)-treated group (0.78 mg/kg) and the WJR-treated group (22.9 g/kg). A group of 10 healthy rats was used as normal control. Treatments or normal saline for the control group were administered by oral gavage once daily. Rats were sacrificed after 30-day treatment and subjected to the following examinations: arthritis index (AI) was estimated, inflammatory cell infiltration and proliferation in synovial membrane were evaluated by microscopy, the synoviocyte apoptosis was determined by TUNEL assay, and the cell apoptosis index was calculated. Results: AI was lowered significantly in the WJR group compared to the model group (P<0.01). The pathological findings observed in the model group were reversed in the WJR group, including increase in inflammatory cell infiltration and synoviocyte proliferation in synovial membrane and reduction in cell apoptosis index (all P<0.01). Conclusions: Synoviocyte proliferation and apoptosis reduction were present in CIA rats. WJR was effective in treating the rat model of CIA. The therapeutic effect might be exerted through inducing apoptosis and suppressing proliferation of synoviocytes.     

Role of β2-adrenoceptor-β-arrestin2-nuclear factor-κB signal transduction pathway and intervention effects of oxymatrine in ulcerative colitis

Objective:To investigate the β2-adrenoceptor(β2AR)-β-arrestin2-nuclear factor-κB(NF-κB) signal transduction pathway and the intervention effects of oxymatrine in a rat model of ulcerative colitis.Methods: Forty SD rats were randomly divided into four groups,which included the normal control group,the model group, the mesalazine group and the oxymatrine treatment group,with 10 rats per group.Experimental colitis induced with trinitrobenzene sulfonic acid(TNBS) was established in each group except the normal control group.The rats in the oxymatrine treatment group were treated with intramuscular injection of oxymatrine 63 mg/(kg·d) for 15 days and the rats in the mesalazine group were treated with mesalazine solution 0.5 g/(kg·d) by gastric lavage for 15 days. The rats in the normal control group and model group were treated with 3 mL water by gastric lavage for 15 days. Diarrhea and bloody stool were carefully observed.Histological changes in colonic tissue were examined on day 7 in 2 rats per group that were randomly selected.The expression of β2AR,β-arrestin2 and NF-κB p65 in colon tissue and spleen lymphocytes were detected with immunohistochemistry and Western immunoblotting techniques on day 16 after fasting for 24 h.Six rats died of lavage with 2 each in the normal control,the model group and the mesalazine group;and were not included in the analysis.Results:The rats in the model group suffered from looser stool and bloody purulent stool after modeling.But in the oxymatrine and mesalazine groups,looser stool and bloody purulent stool reduced after treatment.And the colonic wall in the model group was thickened and the colon length shortened.The colon mucosa was congested in multiple areas with edema,erosion,superficial or linear ulcer and scar formation,while the intestinal mucosa injury reduced in the mesalazine and oxymatrine groups(P<0.01).In colonic mucosa and in spleen lymphocytes,compared with the normal control group,the expression of NF-κBp65 were significantly increased(P<0.01) in the model group while the expressions ofβ2AR andβ-arrestin2 were significantly decreased(P<0.01).Compared with the model group,the expression of NF-κBp65 was significantly decreased in the mesalazine group(P<0.01) and oxymatrine treatment group(P<0.01) while the expressions of β2AR and β-arrestin2 were significantly increased(P<0.01).There were no statistically significant differences in the expression of β2AR,β-arrestin2 and NF-κBp65 between the mesalazine group and oxymatrine group(P>0.05).Conclusions:The β2AR-β-arrestin2-NF-κB signal transduction pathway participated in the pathologic course of ulcerative colitis.Oxymatrine attenuated ulcerative colitis through regulating the β2AR-β-arrestin2-NF-κB signal transduction pathway.     

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.     

Synergistic Anti-inflammatory Effect of Radix Platycodon in Combination with Herbs for Cleaning-heat and Detoxification and Its Mechanism

Objective:To investigate the synergistic anti-inflammatory effect of Radix Platycodon in combination with herbs for cleaning-heat and detoxification and its mechanism for Fei(肺)-targeting.Methods: Forty Wistar rats were randomly divided into five groups(8 per group):the sham-operated group,model group, Radix Platycodon group,Flos Lonicera and Fructus Forsythia(LF) group,and Radix Platycodon,Flos Lonicera and Fructus Forsythia combination(PLF) group,using a random number table.A rat chronic obstructive pulmonary disease(COPD) model was established by passive smoking and intratracheal instillation of lipopolysaccharide(LPS).The treatments started from the 15th day of passive smoking for a total duration of 14 days.At the end of the treatment,changes in the following measurements were determined:lung histopathology, inflammatory cytokines including tumor necrosis factorα(TNF-α),transforming growth factorβ(TGF-β) and interleukin IL-1β(IL-1β) in bronchoalveolar lavage fluid(BALF),and mRNA expression of endogenous active substance intestinal trefoil factor 3(TFF3) in the lung tissue.Results:Light microscopy showed that compared with the sham-operated group,rats in the COPD model group had disrupted alveolar structure,collapsed local alveoli,significantly widened or even fused alveolar septa,and massive infiltration of inflammatory cells in the alveolar wall and interstitium.In addition,significant bronchial epithelium hyperplasia,partially shed epithelia, and marked inflammatory cell infiltration in the bronchial wall and its surrounding tissues were noticed.Electron microscopy showed that rats in the model group had degeneration of alveolar type II epithelial cell;reduction, breakage or even loss of cell surface microvilli;swollen mitochondria with disappearing cristae and vacuole-like structure;and,increased secondary lysosomes in alveolar macrophages.The TNF-α,TGF-βand IL-1βlevels and white blood cell(WBC) count in BALF were significantly increased(P<0.01 or P<0.05) and TFF3 mRNA expression in the lung tissue was significantly reduced(P<0.01).After treatment,the pathological morphology of lung injury was less severe in all three treatment groups.In addition,TGF-βand IL-1βand WBC count in BALF were decreased(P<0.01 or P<0.05),and TFF3 mRNA expression in the lung tissue was significantly increased in the PLF group(P<0.01).Compared with the LF group,the IL-1 p in BALF was significantly decreased (P<0.05),and TFF3 mRNA expression was significantly increased(P<0.05) in the PLF group.Conclusions:Radix Platycodon synergizes with herbs for cleaning-heat and detoxification in reducing inflammatory injury in a rat model of COPD.The synergistic anti-inflammatory effect is reflected in the improvement in pathological changes and in the reduction of IL-1 p levels in BALF.The mechanism of such synergistic action may be related to its effect on maintaining the TFF3 mRNA expression and Fei-targeting function.     

Anti-ulcerogenic activity of aqueous extract of Carica papaya seed on indomethacin-induced peptic ulcer in male albino rats

OBJECTIVE: Carica papaya is an important fruit with its seeds used in the treatment of ulcer in Nigeria. This study investigated the anti-ulcerogenic and antioxidant activities of aqueous extract of Carica papaya seed against indomethacin-induced peptic ulcer in male rats.METHODS: Thirty male rats were separated into 6 groups(A–F) of five rats each. For 14 d before ulcer induction with indomethacin, groups received once daily oral doses of vehicle(distilled water), cimetidine 200 mg/kg body weight(BW), or aqueous extract of C. papaya seed at doses of 100, 150 or 200 mg/kg BW(groups A, B, C, D, E and F, respectively). Twenty-four hours after the last treatment, groups B, C, D, E and F were treated with 100 mg/kg BW of indomethacin to induce ulcer formation. RESULTS: Carica papaya seed extract significantly(P<0.05) increased gastric p H and percentage of ulcer inhibition relative to indomethacin-induced ulcer rats. The extract significantly(P<0.05) decreased gastric acidity, gastric acid output, gastric pepsin secretion, ulcer index and gastric secretion volume relative to group B. These results were similar to that achieved by pretreatment with cimetidine. Specific activities of superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase and glucose-6-phosphate dehydrogenase in the extract-treated groups(D, E and F) were increased significantly over the group B(P<0.05). Pretreatment with the seed extract protected rats from the indomethacin-mediated decrease in enzyme function experienced by the group B. Similarly, indomethacin-mediated decrease in reduced glutathione level and indomethacin-mediated increase in malondialdehyde were reversed by Carica papaya extract. CONCLUSION: In this study, pretreatment with aqueous extract of Carica papaya seed exhibited antiulcerogenic and antioxidant effects, which may be due to the enhanced antioxidant enzymes.     

Influences of electro-acupuncture at related Jing-well Points in rats with vascular dementia

OBJECTIVE:To observe the effects of electro-acupuncture(EA) at related Jing-well Points(HT 9,PC 9,KI 1 and LU 11) in rats with vascular dementia(VD) and discuss the relative mechanism.METHODS:A randomized controlled animal experiment was designed.A total of 104 rats were involved in the present study and divided randomly into 4 groups:sham-operation group,model group,Jing-well Points group,and medication group.The VD model was established according to the modified 4-vessel occlusion(4-VO) method.VD rats in the Jing-well Points group were treated by EA at the related Jing-well Points(HT 9,PC 9,KI 1 and LU 11) while those in the medication group were treated with nimotop.The step-down avoidance test was performed before and after treatment in all rats.Latency and error frequency indexed memory function were recorded.Nitric oxide(NO) levels and superoxide dismutase(SOD) activity in both cerebral tissue and serum were detected after the treatment course.RESULTS:A total of 42 rats were included in the final analysis.Compared with the model group,the latency in the Jing-well Points group was significantly prolonged(P<0.01) and the error frequency was significantly decreased(P<0.05) after therapy;the decrease in NO levels in both brain tissue and serum was significant(P<0.05 and P<0.01,respectively);and the increase in SOD activity was also significant(P<0.01).There was no significant difference in latency,error frequency,NO levels and SOD activity between the Jing-well Points group and medication group.CONCLUSION:EA at related Jing-well Points can remarkably improve memory impairment in VD rats.Moreover,decreasing the overproduction of NO and strengthening the ability of eliminating free radicals may provide therapeutic potential for the treatment of VD.     

Effect of scraping therapy on Interleukin-1 in serum of rats with lumbar disc herniation

OBJECTIVE:To explore the effect of scraping therapy on the Interleukin-1 (IL-1) levels of rats with lumbar disc herniation (LDH). METHODS: Fifty male rats were devided into a blank group (A), a sham operation group (B), a model group (C), a scraping group (D), and a drug group (E). The rats in the group B were treated with sham operation, and groups C, D and E were made into the LDH model by operation. After operation group C were treated with no interventions, D were given scraping and E were fed with azathioprine Then the IL-1 levels of different groups were detected by enzyme-linked immuno sorbent assay method. And the transplanted coccygeal vertebra discs were observed by pathological section. RESULTS: The IL-1 levels in the groups C, D, and E were significantly higher than those in the groups A and B (all P<0.01), which proved the operationwas successful.The IL-1 levels in the groups D and E at different periods had statistical significance (F= 414.158, P<0.01). The treatment periods and interventions have interation (F=46.613, P<0.01). Multiple comparison results showed that the IL-1 levels in the groups D and E was significantly lower than that in the group C (P<0.01), while the IL-1 levels between the groups D and E had no statistical significance (P>0.05). Moreover, pathological section indicated that immuno-inflammatory response was hardly found in coccygeal vertebra discs in the groups A and B, while local immuno-inflammatory responses of the groups D and E were much lighter than that of the group C. CONCLUSION: Scraping therapy could inhibit the immuno-inflammatory responses in the rats with LDH caused by transplantation of autologous nu cleus pulposus.     

Effect of Acupuncture on Proliferation and Differentiation of Neural Stem Cells in Brain Tissues of Rats with Traumatic Brain Injury

Objective:To observe the effect of acupuncture on proliferation and differentiation of neural stem cells in brain tissues of rats with traumatic brain injuny.Methods:Thirty SD rats were randomly and equally allocated to the sham-operated,the model and the acupuncture groups.The traumatic brain injury model was established by the free drop method.For the rats in the acupuncture group,acupuncture was applied once a day for 7 days.Brain histotomy was carried out when treatments were completed.Immunohistochemical techniques were adopted to detect the cells that express nestin,neurofilament proteins(NF)-200 and glial fibrillary acidic proteins(GFAP),the markers of neural stem cells,neurons,astrocytes respectively.Results:Compared to the sham-operated group,the number of nestin-positive cells and NF-200-positive cells in brain tissues was decreased significantly in the model group(P<0.01),whereas the number of GFAP-positive cells was significantly increased (P<0.01).Compared to the model group,the positive cells of nestin,NF-200,GFAP in brain tissues in the acupuncture group were increased obviously(P<0.01).Conclusions:Acupuncture can significantly increase the number of nestin-positive cells,NF-200-positive cells and GFAP-positive cells,indicating the significant increase of neural stem cells,neurons and astrocytes in number.Acupuncture can improve neuranagenesis by promoting the proliferation and differentiation of neural stem cells in brain tissues.This might be one of the mechanisms for acupuncture to treat traumatic brain injury and to promote the repair of nervous function.     

Protective effects and mechanism of total coptis alkaloids on A β25–35 induced learning and memory dysfunction in rats

Objective: To observe the effect of total coptis alkaloids (TCA) on β-amyloid peptide (Aβ25-35) induced learning and memory dysfunction in rats, and to explore its mechanism. Methods: Forty male Wistar rats were randomly divided into four groups: the control group, the model group, the TCA low dose (60 mg/kg) group and the TCA high dose (120 mg/kg) group, 10 in each. Aβ25-35 (5μl, 2μg/μl) was injected into bilateral hippocampi of each rat to induce learning and memory dysfunction. TCA were administered through intragavage for consecutive 15 days. Morris Water Maze test was used to assess the impairment of learning and memory; concentration of malondialdehyde (MDA) in cerebral cortex was determined by thiobarbituric acid reactive substance to indicate the level of lipid peroxidation in brain tissues; activity of manganese-superoxide dismutase (Mn-SOD) in cerebral cortex was determined by xanthine-oxidase to indicate the activity of the enzyme; and NF-κB protein expression in cerebral cortex was measured by SP immunohistochemistry. Results: (1) Morris Water Maze test showed that, during the 4 consecutive days of acquisition trials, the rats in the model group took longer latency and searching distance than those in the control group (P<0.01), which could be shortened by high dose TCA (P<0.05); during the spatial probe trial on the fifth day, the rats in the model group took shorter searching time and distance on the previous flat area than those in the control group (P<0.01), which could be prolonged after TCA treatment (for low dose group, P<0.05; for high dose group, P<0.01). (2) Analysis of cerebral cortical tissues showed that, compared with the control group, MDA level got significantly increased and Mn-SOD activity decreased in the model group (both P<0.01). After having been treated with TCA, the MDA level got significantly decreased (P<0.05 and P<0.01 respectively for low and high dose group), while relative increase of Mn-SOD activity only appeared in high dose group (P<0.05). (3) Immunohistochemistry analysis showed the protein expression of NF-κB got significantly increased after modeling, while high dose TCA can significantly inhibit it. Conclusion: TCA could improve Aβ25-35 induced dysfunction of learning and memory in rats, and its protective mechanism is associated with its actions in decreasing MDA level, increasing Mn-SOD activity and inhibiting the expression of NF-κB in cerebral cortex.     

High-frequency stimulation of anterior nucleus thalamus improves impaired cognitive function induced by intra-hippocampal injection of Aβ1-40 in rats

Background The advent of brain stimulation techniques to treat movement disorders and psychiatric diseases has shown potential to decode the neural mechanism that underlies the cognitive process by modulating the interrupted circuit.Here,the present investigation aimed at evaluating the influence of deep brain stimulation of the anterior nucleus thalamus （ANT-DBS） on memory.Methods Thirty-two rats were randomized into phosphate buffer saline （PBS） group （n=8,rats received PBS injections without implantation of electrodes into the ANT）,Alzheimer＇s dementia （AD） group （n=8,rats received Aβ1-40 injections without implantation of electrodes into the ANT）,ANT sham stimulation group （n=8,rats received Aβ1-40 injections with implantation of electrodes into the ANT but without stimulation） and ANT stimulation group （n=8,rats received Aβ1-40 injections with implantation of electrodes into the ANT and stimulation）.A Morris maze test was used for determining the effect of electrical stimulation on cognitive function in rats.The data were assessed statistically with one-way analysis of variance （ANOVA） followed by Tukey＇s tests for multiple post hoc comparisons.Results The data showed that in the training test,PBS group and AD group managed to learn the hidden-platform faster and faster while AD group needed a significantly longer time to reach the platform than PBS group （P ＜0.05）.Meanwhile,ANT stimulation group demonstrated a significantly shorter time to reach the platform （P ＜0.05） compared to the AD group,while there was no significant difference between the ANT sham stimulation group and the AD group （P ＞0.05）.On the probe test,the AD group spent less time （（10.15±2.34） seconds） in the target quadrant than the PBS group （（28.20±2.75） seconds） （P ＜0.05）.And the times of platform-traversing of the AD group （3.35±1.12） significantly decreased compared with the PBS group （8.69±2.87） （P ＜0.05）.However,the times of platform-traversing and the time spent in the target quadrant of the ANT stimulation group significantly increased compared to the AD group （P ＜0.05）,while times of platformtraversing or the time spent in the target quadrant was not significantly different between the ANT sham stimulation group and the AD group （P ＞0.05）.Conclusion Bilateral high-frequency stimulation of the ANT may be useful as a potential therapeutic modality for cognitive dysfunction in AD.     

Role of DOR-β-arrestinl-Bcl2 Signal Transduction Pathway and Intervention Effects of Oxymatrine in Ulcerative Colitis

This study was aimed to investigate the role of the delta-opioid receptor （DOR）-β-arrestinl-Bcl-2 signal transduction pathway in the pathogenesis of ulcerative colitis （UC） and the intervention effects of oxymatrine on UC. Forty Sprague-Dawley rats were divided into nor- mal group, model group, oxymatrine-treated group and mesalazine-treated group （n=10 each） at ran- dom. The rat UC model was established by intra-colonic injection of trinitrobenzene sulfonic acid in the model group and two treatment groups. The rats in oxymatrine-treated group were subjected to intramuscular injection of oxymatrine [63 mg/（kg·day）] for 15 days, and those in mesalazine-treated group given mesalazine solution [0.5 g/（kg·day）] by gastric lavage for the same days. Animals in normal group and model group were administered 3 mL water by gastric lavage for 15 days. On the 16th day, after fasting for 24 h, the rats were sacrificed for the removal of colon tissues. The expres- sion levels of DOR, β-arrestinl and Bcl-2 were determined in colon tissues by immunohistochemistry and real-time quantitative polymerase chain reaction （RT-PCR）, respectively. It was found that the expression levels of DOR, [3-arrestinl and Bcl-2 protein and mRNA were significantly increased in the model group as compared with the other groups （P〈0.05）. They were conspicuously decreased in both mesalazine-treated and oxymatrine-treated groups in contrast to the model group （P〈0.05）. No statistically significant difference was noted in these indices between mesalazine- and oxyma- trine-treated groups （P〉0.05）. This study indicated that the DOR-β-arrestinl-Bcl-2 signal transduc- tion pathway may participate in the pathogenesis of UC. Moreover, oxymatrine can attenuate the de- velopment of UC by regulating the DOR-β-arrestin 1-Bcl-2 signal transduction pathway.     

Effects of Osthole on the Right Ventricle Remodeling in Monocrotaline-Treated Rats

Objective:To investigate the effects of Osthole(Ost) on the right ventricle remodeling in monocrotaline-treated rats,and to explore the mechanisms. Methods:200~220 g male Sprague-Dawley rats were randomly divided into normal control group(Control,n=8),model group(Model,n=8),low dose of Ost treatment group(Ost-L,10 mg·kg~(-1),n=8),high dose of Ost treatment group(Ost-H,20 mg·kg~(-1),n=8) and sildenafil treatment group(Sildenafil,25 mg·kg~(-1),n=8).All rats were given a single dose of MCT 50 mg·kg~(-1) subcutaneously to establish the right ventricle remodeling except normal control group. Then the rats in the Ost and sildenafil treatment group were gavaged once daily from 1 day to 28 days. The other rats in the model group and normal control group were given the same amount of dd H_2O with 0.5% be tween 80. After 28 days of administration,the right ventricular pressure were measured by right heart catheterization. Right ventricle(RV) and left ventricle plus septum(LV+SEP) were weighed separately. RV hypertrophy index(RVHI) were measured by the relative weight ratio of RV to LV+SEP. The morphological change of the RV were executed by light microscope and transmission electron microscope.The cardiomyocytes apoptosis were detected by Td T mediated nick d UTP end-labeling(TUNEL) method using the paraffin-embedded right ventricular tissues. The m RNA expression of Bcl-2,Bok and p53 were examined by real time RT-PCR. The protein expression of Bcl-2,Bax,Bok,IκB,IL-6,TNF-α and the levels of Cleaved caspase 3,p-NF-κB(p65) were detected by western blotting. Results:Compared with the control group,the right ventricular pressure and RVHI were increased obviously(P<0.05),myocardial hypertrophy,structure disorders,mitochondrial swelling and cardiomyocytes apoptosis(P<0.05) were observed in model group. The m RNA expression of Bok and p53 in right ventricular tissues were up-regulated(P<0.05),and the m RNA expression of Bcl-2 was down-regulated in model group(P<0.05).The protein expression of Bax,Bok,IL-6,TNF-α and the levels of Cleaved caspase 3,p-NF-κB(p65) were up-regulated(P<0.05),and the protein expression of Bcl-2 and IκB were down-regulated significantly in model group(P<0.05). Compared with the model group,the right ventricular pressure and RVHI were decreased(P<0.05),myocardial hypertrophy,structure disorders,mitochondrial swelling and cardiomyocytes apoptosis(P<0.05) were improved significantly in Ost and sildenafil treatment group. The m RNA expression of Bok and p53 were downregulated(P<0.05),while the m RNA expression of Bcl-2 was up-regulated(P<0.05) in Ost and sildenafil treatment group. The protein expression of Bax,Bok,IL-6,TNF-α,and the levels of Cleaved caspase 3,p-NF-κB(p65) were down-regulated(P<0.05),and the protein expression of Bcl-2 and IκB were upregulated significantly in Ost and sildenafil treatment group(P<0.05). Conclusions:Ost can resist the RV remodeling induced by MCT in rats,which may be related to these possible mechanisms:(1) Ost inhibits cardiomyocytes apoptosis by regulating Bax/Bcl-2 pathways;(2) Ost attenuates inflammation by inhibiting NF-κB pathway.