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

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

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.     

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.     

Effect of Quyu Chencuo Formula(去菀陈莝方)on Renal Fibrosis in Obstructive Nephropathy Rats

Objective: To observe the effect of Quyu Chencuo Formula(去菀陈莝方, QCF) on renal fibrosis in rats with obstructive nephropathy. Methods: Twenty-four rats were randomly divided into three groups, 4 for sham operation as the control group, 10 for unilateral ureteral obstruction(UUO) model group, and the rest 10 for QCF treating UUO model group. All rats were sacrificed under 3% pentobarbital(50 mg/kg) anesthesia on the 14 th day after surgery, then the right kidney samples of rats were harvested for hematoxylin eosin(HE) staining and Masson staining to observe the renal pathological changes. Immunohistochemistry and Western blotting were used to examine the expression of transforming growth factor β1(TGF-β1), and real-time polymerase chain reaction(RT-PCR) was employed to examine the expressions of TGF-β1, α-smooth muscle actin(α-SMA) and E-cadherin mRNA. Results: HE and Masson staining showed that the renal interstitial of the rats in the control group had no significant fibrotic lesion; in the model group, there were obvious interstitial fibrosis; for the QCF group, there were epithelial cell necrosis, infiltration of lymphocytes and mononuclear cells, aggravated interstitial fibrosis in varied degrees, but the pathological changes were less in the QCF group than in the model group. The immunohistochemistry and Western blotting results showed that the TGF-β1 expression was increased significantly in the model group, while decreased significantly in the QCF group(P0.05); RT-PCR showed that the mRNA expression of α-SMA and TGF-β1 increased significantly in the model group, while both were significantly decreased in the QCF group compared with the model group(P0.05). The mRNA expression of E-cadherin was decreased significantly in the model group, and it was significantly increased in the QCF group as compared with the model group(P0.05). Conclusion: QCF may improve renal fibrosis by regulating the expressions of TGF-β1, α-SMA and E-cadherin, and prevent the progress of kidney fibrosis.     

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.     

Effect of Wenhua Juanbi Recipe (温化蠲痹方) on Expression of Receptor Activator of Nuclear Factor Kappa B Ligand, Osteoprotegerin, and Tumor Necrosis Factor Receptor Superfamily Member 14 in Rats with Collagen-Induced Arthritis

Objective: To study the effect of Wenhua Juanbi Recipe(温化蠲痹方, WJR) on expression of receptor activator of nuclear factor kappa B ligand(RANKL), osteoprotegerin(OPG), and tumor necrosis factor receptor superfamily member 14(TNFRSF14, also known as LIGHT) in rats with collagen-induced arthritis(CIA). Methods: CIA rats were generated by subcutaneous injection of bovine collagen type-Ⅱ at the tail base. Sixty CIA rats were randomly assigned(10 animals/group) to: model, methotrexate(MTX)-treated(0.78 mg/kg body weight), and WJR-treated(22.9 g/kg) groups. Healthy normal rats(n=10) were used as the normal control. Treatments or saline were administered once daily by oral gavage. Rats were sacrificed at day 28 post-treatment and knee synovium and peripheral blood serum were collected. Toe swelling degree and expression of RANKL, OPG, and LIGHT were determined by Western blot and immunohistochemistry. Results: Compared with the normal group, toe swelling degree was significantly increased in the model group(P0.01). After treatment, toe swelling degree decreased significantly in the WJR and MTX groups compared with the model group(P0.01). Compared with the normal group, expression of RANKL and LIGHT were significantly increased and OPG significantly decreased in peripheral blood and synovium of the model group(P0.01). Conversely, RANKL and LIGHT expression were significantly reduced and OPG increased in the WJR and MTX groups compared with the model group(P0.01). No statistically significant difference existed between WJR and MTX groups. Conclusion: WJR likely acts by reducing RANKL expression and increasing OPG expression, thus inhibiting RANKL/RANK interaction and reducing LIGHT expression, thereby inhibiting osteoclast formation/activation to block bone erosion.     

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.     

Xuezhikang(血脂康) Reduced Renal Cell Apoptosis in Streptozocin-Induced Diabetic Rats through Regulation of Bcl-2 Family

Objective: To investigate the effect of Xuezhikang(血脂康, XZK) on renal cell apoptosis in diabetic rats and the possible mechanism. Methods: Sixty-six rats were randomly divided into 3 groups: the normal, model and XZK groups. In each group, the rats were further randomly divided into 3-month and 6-month subgroups, respectively. Diabetes of rats was induced by a single intraperitoneal injection of 1% streptozocin at 60 mg/kg body weight. Rats in the XZK group received gastric perfusion of XZK(1200 mg/kg body weight) everyday for 3 or 6 months, while rats in the normal and model groups received equal volume of saline. Twenty-four hours' urine was collected for urinary albumin excretion(UAE) measurement. Periodic acid-Schiff(PAS) and Masson's trichrome staining were used for saccharides and collagen detection. Cell apoptosis of renal cortex was investigated by Td T-mediated d UTP nick end labeling(TUNEL) staining. Bax and Bcl-2 expressions were detected by immunohistochemistry and Western blot, respectively. Cytochrome C(Cyt C) and caspase-9 concentration were detected by Western blot. Results: Compared with the model group, XZK treatment could significantly decrease the kidney hypertrophy index, 24 h UAE, renal cell apoptosis, cytoplasmic Cyt C level and active caspase-9 level, as well as suppress the increment of Bax and up-regulate the expression of Bcl-2, leading to the suppression of Bax/Bcl-2 ratio at 3 and 6 months(P0.05 or P0.01). Moreover, XZK treatment could alleviate the deposition of PAS-stained saccharides and Masson's trichromestained collagen to different extent. Conclusions: Renal cell apoptosis was observed in diabetic kidney, in which mitochondrial apoptotic pathway might be involved. XZK treatment could attenuate pathological changes in diabetic kidney and reduce renal cell apoptosis, probably via the suppression of Bax/Bcl-2 ratio, which lead to inhibition of Cyt C release and following caspase-9 activation.     

丁苯酞对阿尔茨海默病模型大鼠P38及ERK表达的影响及意义

目的 研究阿尔茨海默病模型大鼠P38、ERK的表达及丁苯酞对其影响和意义.方法 成年雄性SD大鼠60只,随机分为模型组、丁苯酞组及对照组,每组20只;通过向大鼠海马内注射Aβ_(1-42)建立阿尔茨海默病模型,喂养60d后用Y型迷宫检测大鼠学习记忆能力、免疫组织化学染色检测大鼠脑组织P38和ERK的表达.结果 与对照组比较,模型组大鼠学习记忆能力下降,其海马CA1区P38阳性细胞增多,皮层ERK表达减少,差异均有统计学意义(P＜0.01);与模型组比较,丁苯酞组大鼠海马Ca1区P38阳性细胞数明显减少,皮层ERK表达增多,差异有统计学意义(P<0.01).结论 丁苯酞改善阿尔茨海默病大鼠学习记忆能力可能与其抑制P38、增加ERK的表达有关.

Abstract：

Objective To determine the effect of butylphthalide on the expressions of p38 mitogen-activated protein kinase and extra-cellular signal regulated kinases (ERKs) in the brain tissue of rats with Alzheimer's disease (AD). Methods Sixty male adult rats were randomly divided to AD model group, butylphthalide group, and control group (n=20). AD models were established by injecting beta-amyloid protein 1 -42 into the hippocampus of rats. Sixty days later, the learning and memory abilities of the rats were evaluated using Y-maze test, and the expressions of p38 and ERKs in the brain tissue of the rats were measured by immunohistochemistry. Results Compared with the control group, the rats in AD model group exhibited significantly reduced learning and memory abilities, increased expressions of P38 in the hippocampus and lowered expression of ERK in the cortex (P<0.01). In comparison with the model group, the rats in the butylphthalide group showed significantly decreased P38-positive cells in the hippocampus and increased expression of ERK in the cortex (P<0.01). Conclusion Butylphthalide improves the learning and memory abilities of rats with experimental AD, the mechanism of which may involve inhibition of P38 expression and enhancement of ERK expression in the brain tissues.     

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 exercise on behavior and peripheral blood lymphocyte apoptosis in a rat model of chronic fatigue syndrome

This study examined the effects of exercise on behavior and peripheral blood leukocyte apoptosis in a rat model of chronic fatigue syndrome （CFS）. Thirty-six healthy male Sprague-Dawley rats were equally randomized into 3 groups： the control group, CFS model group and the exercise group in terms of body weight. A total of 25 rats entered the final statistical analysis due to 11 deaths during the study. CFS model was established by subjecting the rats in CFS model group and exercise group to electric shock, chronic restraint stress and cold water swim. Besides, rats in the exercise group took rtmning wheel exercise. After a week of conditioning feeding, model construction and running wheel exercise were performed simultaneously, and lasted for 23 consecutive days. The behavior experiments, including running wheel exercise, open-field test, tail suspension test and Morris water maze test, were conducted, either before or after the model establishment. Rats were sacrificed and peripheral blood was obtained for the assessment of lymphocyte apoptosis index by flow cytometry （FCM）. It was found that as compared with those in the control group, the weight of the rats was decreased obviously （P〈0.01）, the mobility time in the open-field and the tail suspension tests was shortened significantly （P〈0.01）, the time to locate the platform was enhanced （P〈0.01） and the cell apoptosis index was increased substantially （P〈0.01） in the CSF model group. Meanwhile, in comparison to the model group, the behavior in the open-field and the tail suspension tests was improved significantly （P〈0.05）, and the apoptosis index decreased remarkably （P〈0.01） in the exercise group. It is concluded that sport intervention can prevent lymphocyte apoptosis and improve animal behavior rather than the memory.     

Effects of Zuogui Pill(左归丸) and Yougui Pill(右归丸) on the Expression of Brain-Derived Neurotrophic Factor and Cyclic Adenosine Monophosphate/Protein Kinase A Signaling Transduction Pathways of Axonal Regeneration in Model Rats with Experimental Autoimm

Objective:To study the effects of Zuogui Pill(左归丸,ZGP)and Yougui Pill(右归丸,YGP)on the expressions of brain-derived neurotrophic factor(BDNF)and cyclic adenosine monophosphate(cAMP)/protein kinase A(PKA)signaling of axonal regeneration in the Lewis rats with experimental autoimmune encephalomyelitis(EAE),in order to explore the possible mechanism of ZGP and YGP on promoting axonal regeneration.Methods:The rats were randomly divided into normal control(NC),model(MO),prednisone acetate(PA),ZGP and YGP groups.The EAE model of rat was established by injecting antigen containing myelin basic protein(MBP)_(68-86).The brain and spinal cord were harvested on the 14th and 28th day postimmunization(PI),the protein and mRNA expression of BDNF and PKA in the brain and spinal cord of rats were detected by Western blot analysis and real-time quantitative polymerase chain reaction(PCR),and the cAMP levels were detected by using enzyme-immunoassay method.Results:(1)On the 28th day PI,the mRNA expression of BDNF in brain white matter and spinal cord of rats in ZGP and YGP groups were up-regulated,especially in YGP group(P0.05 or P0.01).(2)On the 14th day PI,the cAMP levels in brain white matters significantly increased in PA and YGP groups compared with MO group(P0.05 or P0.01),and the cAMP level in YGP group was higher than that in ZGP group(P0.05).The cAMP level in spinal cord also significantly increased in YGP group compared with MO,PA and ZGP groups,respectively(P0.01).(3)On the 14th day PI,the PKA expression in spinal cord of rats in ZGP group was significantly decreased compared with MO and YGP groups,respectively(P0.05).(4)On the 28th day PI,there was a positive correlation between cAMP and PKA expression in the brain white matter of YGP rats.Conclusions:The results suggest that ZGP and YGP may promote axonal regeneration by modulating cAMP/PKA signal transduction pathway,but the targets of molecular mechanism of ZGP may be different from those of YGP.     

Effects of Acupuncture on Progesterone and Prolactin in Rats of Embryo Implantation Dysfunction

Objective:To investigate the effect of acupuncture on progesterone(P4) and prolactin(PRL) in rats of embryo implantation dysfunction(EID).Methods:On the first day of pregnancy,72 female Wistar rats were randomly allocated into the normal group,the EID model group,the acupuncture group and the P4 group(18 in each group).The normal group was injected sesame oil,while the other three groups were given mifepristone to establish the EID model.The acupuncture group and the P4 group were given treatment of acupuncture and P4 injection,respectively.The serum of P4 and PRL were detected by radioimmunoassay,and the mRNA and protein expressions of P4 receptor(PR) and PRL receptor(PRLR) were detected using real-time polymerase chain reaction and immunohistochemical method,respectively.Results:Compared with the normal group,the serum levels of P4 and PRL as well as the mRNA and protein expression levels of PR and PRLR in the EID model group were significantly lowered(P<0.01 or P<0.05).The above indices in the acupuncture group and the P4 group were significantly elevated compared with the EID model group(P<0.01 or P<0.05).Conclusion:Acupuncture can promote embryo implantation effectively,which might be related to the effects of acupuncture on upregulating the P4 and PRL levels in serum and the PR and PRLR expression levels in rats.     

加味竹叶石膏汤及其成分可预防大鼠放射性食管炎的发生

Objective:To investigate the effect of Modified Zhuye Shigao Decoction(加味竹叶石膏汤,MZSD)and its components on preventing radiation esophagitis of rats.Methods:One hundred Wistar rats were randomly divided into 5 groups,including the control group,radiation model group,MZSD group,Zhuye Shigeo Decoction(竹叶石膏汤,ZSD)group,and added Ingredients group,20 rats in each group.The model of radiation esophagitis of rat was established by once local radiation of 40 Gy(330 Mulmin)with a high energy linear accelerator.The administration of Chinese medicine was continued for 14 days from 7 days before radiation application in the three treatment groups.On the 7th and 14th day,the serum was isolated and the levels of inflammatory cytoldnes tumor necrosis factor(TNF-α),interleukin 1 13(IL-1β)and IL-8 were tested.The pathological slices of esophagus were obtained,and the pathological changes were observed.During the whole process,weight and food intake were recorded each day.Results:On the 7th day after radiation,the esophagus of rats in the MZSD group was almost intact,and the pathological injury score was significantly lower than that of the radiation model group,ZSD group and added ingredients group(P0.01).Compared with the control group,the body weight and food intake of rats in the radiation model group were significantly decreased,and the levels of TNF-α,IL-1β and IL-8 were significantly increased(P0.05 or P0.01),while the MZSD group showed a significant increase in body weight and food intake,and a significant decrease in the levels of TNF-α,IL-1β and IL-8 compared with the radiation model group,ZSD group and added ingredients group(P0.05 or P0.01).Conclusion:MZSD prevents the development of radiation esophagitis probably by inhibiting the generation and release of the inflammatory cytoldnes TNF-α,IL-1β and IL-8.     

Effect of Soothing Gan (Liver) and Invigorating Pi (Spleen) Recipes on TLR4-p38 MAPK Pathway in Kupffer Cells of Non-alcoholic Steatohepatitis Rats

Objective: To investigate the mechanism of inflammatory-mediated toll-like receptor 4(TLR4)-p38 mitogen-activated protein kinase(p38 MAPK) pathway in Kupffer cells(KCs) of non-alcoholic steatohepatitis(NASH) rats and the intervention effect of soothing Gan(Liver) and invigorating Pi(Spleen) recipes on this pathway. Methods: After 1 week of acclimatization, 120 Sprague-Dawley male rats were randomly divided into 8 groups using a random number table(n=15 per group): normal group, model group, low-dose Chaihu Shugan Powder(柴胡疏肝散, CHSG) group(3.2 g/kg), high-dose CHSG group(9.6 g/kg), low-dose Shenling Baizhu Powder(参苓白术散, SLBZ) group(10 g/kg), high-dose SLBZ(30 g/kg) group, and low-and highdose integrated recipe(L-IR, H-IR) groups. All rats in the model and treatment groups were fed with a high-fat diet(HFD). The treatments were administrated by gastrogavage once daily and lasted for 26 weeks. The liver tissues were detected with hematoxylin-eosin(HE) and oil red O staining. Levels of liver lipids, serum lipids and transaminases were measured. KCs were isolated from the livers of rats to evaluate the mRNA expressions of TLR4 and p38 MAPK by real-time fluorescence quantitative polymerase chain reaction, and proteins expressions of TLR4, p-p38 MAPK and p38 MAPK by Western blot. Levels of inflammatory cytokines including tumor necrosis factor α(TNF-α), interleukin(IL)-1 and IL-6 in KCs were measured by enzyme-linked immunosorbent assay. Results: After 26 weeks of HFD feeding, HE and oil red O staining showed that the NASH model rats successfully reproduced typical pathogenesis and histopathological features. Compared with the normal group, the model group exhibited significant increases in body weight, liver weight, liver index, serum levels of total cholesterol(TC), triglyceride(TG), low-density lipoprotein cholesterol, and aspartate aminotransferase as well as TC and TG levels in liver tissues, and significant decrease in serum level of high-density lipoprotein cholesterol(P0.05 or P0.01), while those indices were significantly ameliorated in the H-IR group(P0.05 or P0.01). Higher levels of TNF-α, IL-1 and IL-6 in KCs were observed in the model group compared with the normal group(P0.01). Significant decreases in TNF-α, IL-1 and IL-6 were observed in the H-SLBZ, H-IR and L-IR groups compared with the model group(P0.05 or P0.01). The m RNA expressions of TLR4 and p38 MAPK and protein expressions of TLR4, p38 MAPK and p-p38 MAPK in KCs in the model group were significantly higher than the normal group(P0.01), while those expression levels in the L-IR and H-IR groups were significantly lower than the model group(P0.05 or P0.01). Conclusions: Inflammation in KCs might play an important role in the pathogenesis of NASH in rats. The data demonstrated the importance of TLR4-p38 MAPK signaling pathway in KCs for the anti-inflammatory effect of soothing Gan and invigorating Pi recipes.     

Antioxidant Mechanism of Xiaojin Pill(小金丸) for Treatment of Peyronie's Disease in Rats Based on Matrix Metalloproteinases

Objective: To evaluate the effects of Xiaojin Pill(小金丸) in the treatment of Peyronie's disease(PD) in a rat model. Methods: Twenty-four male Sprague-Dawley rats were randomly divided into four groups with 6 in each: sham operation, PD model, vehicle control and Xiaojin Pill groups. The rats in the sham operation group received penile tunica albsginea(TA) injection with 50 μL vehicle, while the rats in the other 3 groups received 50 μL penile TA injection of 50 μg transforming growth factor(TGF)-β1. Forty-two days after the injection, rats in the vehicle control and Xiaojin Pill groups received 0.5 mL water and Xiaojin Pill solution(107 mg/kg of body weight), respectively by gavage for 28 days, while those in the sham operation and PD model groups did not receive any intervention. After intervention, the expressions of matrix metalloproteinase 2/9(MMP2/9), nitric oxidesynthase(NOS), superoxide dismutase(SOD) and malondialdehyde(MDA) were measured. Results: Rats in the PD model and vehicle control groups presented obvious fibrosis in corpus cavernosum(CC) and demonstrated a significantly increased expressions of MMP2 and MMP9 in the CC compared with the sham operation group(all P0.01). In contrast, the expressions of MMP2 and MMP9 in the Xiaojin Pill group were significantly down-regulated(both P0.01). In addition, the levels of NOS and MDA in CC were significantly increased while the activity of SOD was decreased in the PD model and vehicle control groups compared with the sham operation group(al P0.01). After Xiaojin Pil treatment, the levels of MDA, NOS and SOD appeared to be corrected(al P0.01). Conclusions: Xiaojin Pill could reduce fibrosis in the CC by decreasing the expressions of MMPs, NOS and MDA, and by increasing the activity of SOD. Therefore, Xiaojin Pill might be a therapeutic option for PD.     

Effect of Lirukang Oral Liquid(利乳康口服液) on Neurotransmitters and Estrogen in Rats with Hyperplasia of Mammary Gland

Objective: To investigate the effects of Lirukang oral liquid (LRK, 利乳康口服液) on release of neurotransmitter in rats with hyperpiasia of mammary glands (HMG) and to explore its mechanism. Methods: Sixty rats were divided into six groups, the normal control group, the model control group, the large dosage (3.6g/kg) and the small dosage (1.8g/kg) LRK groups, the Ruzengning (乳增宁, RZN, 2.5g/kg) group and the tamoxifen (TAM, 5mg/kg) group, 10 in each group. Except those in the normal control group, all the animals were made into rat model of HMG by intraperitoneal injection of estradiol benzoate. Levels of dopamine (DA) and 5-hydroxytryptamine (5-HT) in hypothalamus and mammary gland in rats were detected by fluorescence luminosity assay, and level of prolactin (PRL) in serum was detected by radioimmunoassay.Results: In the model group, the level of DA reduced significantly (P＜0.01), and 5-HT and PRL increased obviously ( P＜0.01). Compared with the model group, the LRK groups of both dosages and the TAM group had their level of DA significantly increased (P ＜ 0. 01 ), and level of 5-HT significantly decreased ( P＜0.01). The serum PRL in both LRK groups was significantly decreased ( P＜0.01). No obvious changes in DA, 5-HT and PRL were found in the RZN group. Conclusion: LRK and TAM have similar effects in regulating the release of neurotransmitter in hypothalamus and mammary gland and serum content of estrogen in the animal models of HMG.