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 Wuziyanzong Pill on sperm quality and calcium ion content in oligoasthenospermia rats

OBJECTIVE:To study the effect of Wuziyanzong treatment on the sperm quality and content of calcium ions(Ca 2+) in oligoasthenospermia rats.METHODS:A model of oligoasthenospermia was induced in 50 Sprague Dawley rats by treatment with tripterygium glycosides at 30 mg/kg per day for 8 weeks.They were divided randomly into a model group,a positive group(Huangjingzanyu capsule,3.01 g/kg),and low,medium and high dose Wuziyanzong treatment groups(2.30,4.60,9.20 g/kg crude drug respectively) with 10 in each group.Another 10 rats were used as a control group.The rats in the control and model groups were administered distilled water,while the rats in the remaining groups were administered Wuziyanzong for 30 d.The epididymides were removed,spermatozoa recovered and the sperm density and viability were measured.The spermatozoa were purified and the contents of Ca 2+ in the cytoplasm and mitochondria were detected by flow cytometry and atomic absorption spectrometry,respectively.RESULTS:After 8 weeks of treatment with tripterygium glycosides,the sperm density,sperm activity and the Ca 2 + content of spermatozoa in the model rats were all significantly decreased compared with the control group(all P<0.05).After 30 d treatment,the sperm density and activity improved and the Ca 2 + content of sperm were increased significantly in the medium and high dose Wuziyanzong treatment groups in comparison with the model group(all P<0.05).CONCLUSION:The Wuziyanzong treatment increased sperm density,improved sperm viability and enhanced the content of Ca 2+ in the sperm cytoplasm and mitochondria in this rat model of oligoasthenospermia.     

Effects of Kangquan Recipe (康泉方) on sex steroids and cell proliferation in rats with benign prostatic hyperplasia

Objective:To investigate the effects of Kangquan Recipe(康泉方,KQR)on sex steroids and cell proliferation in an experimental benign prostatic hyperplasia(BPH)model in rats.Methods:Seventy-two SD rats were randomly divided into six groups:the normal group,the model group,the finasteride group,and the low-, middle-,and high-dose KQR groups,12 in each group.Except those in the normal group,the rats were injected with testosterone after castration for the establishment of BPH model and then given respectively w...     

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.     

Anti-diabetic Activity of Zhenqing Recipe and Ligustri Lucidi Fructus in Type 2 Diabetic Rats

Objective To investigate the influence of Zhenqing Recipe (ZQR) and Ligustri Lucidi Fructus (LLF) on diabetic rats and its possible mechanism. Methods The model of type 2 diabetic rats was established by feeding a high-sucrose-high-fat diet and injecting a low dose of Streptozotocin in Wistar rats. The model rats were randomly divided into three groups: diabetic model, ZQR-treated, and LLF-treated groups for 8-weeks treatment. The normal Wistar rats were as a normal control group. Results The level of fasting blood glucose in ZQR and LLF groups was decreased compared with model group (P < 0.01, 0.05, respectively). Both ZQR and LLF markedly reduced serum triglycerides (P < 0.01, 0.05, respectively), and increased the insulin sensitivity index (P < 0.05). Histopathology revealed that ZQR and LLF reduced pancreatic damage. Immunohistochemistry evaluation showed that the percentage of insulin positive cells in pancreatic island was higher than model group (P < 0.01, 0.05, respectively).The mRNA and protein expression of SREBP-1c in pancreas were significantly decreased in ZQR and FLL group (P < 0.01). Conclusion ZQR has therapeutic effect on type 2 diabetes, it ameliorates the histopathological changes of pancreas, protects β cells, improves insulin resistance, and attenuates the expression of SREBP-1c. This study also provides the anti-diabetic evidence of FLL even its effects are weaker than ZQR.     

Time-dependent Expression of PEDF and VEGF in Blood Serum and Retina of Rats with Oxygen-induced Retinopathy

The effects of the balance changes of pigment epithelium growth factor(PEDF) and vascular endothelial growth factor(VEGF) in whole-body and retinal tissue on rats with oxygen-induced retinopathy were investigated. Forty-eight neonatal SD rats at the age of 7 days were randomly divided into 4 groups. The neonatal rats in experimental groups were exposed to 75% to 80% oxygen for 5 days and then to normal air, and those in control groups were kept feeding in normal air. At the age of 17 and 22 days, all the neonatal rats received retina angiography with FITC-dextran and the pathological changes of retinal vessels and perfusion were observed. HE staining of the tissue section and the number counting of endothelial cells extending beyond the inner limiting membrane were performed to evaluate the endothelial proliferation. Immunohistochemistry was applied to detect the expression of PEDF and VEGF in retinal tissue, and ELISA to detect their expression in serum. A hypoxic-ischemic proliferation of retina and more endothelial cells extending beyond the inner limiting membrane were found in the neonatal rats in both experimental groups of 17-day old and 22-day old as compared with those in control group with the difference being statistically significant(P<0.01). VEGF staining of the rats in the 17-day old experimental group was significantly stronger, with an increasing positive rate, than that of the rats in the 17-day old control group(P<0.01). PEDF staining of the rats of 22 days old was weaker than that of the rats of 17 days old in the experimental groups(P<0.01). There was no significant difference in serum VEGF concentration among all groups(P>0.05). The serum PEDF concentration in the rats of 17 days old in experimental group was decreased significantly as compared with that in the rats of 17 days old in control group(P<0.01), and in experimental groups, the serum PEDF concentration of the rats of 22 days old was increased as compared with that of the rats of 17 days old(P<0.01). In conclusion, the obviously decreased serum PEDF concentration and the abnormal enhanced expression of VEGF density in local retinal tissue broke down the balance of PEDF/VEGF in whole-body or local tissues, which might play an important role in retinal vascular proliferation.     

Effects of different therapeutic methods and typical recipes of Chinese medicine on activation of c-Jun N-terminal Kinase in Kupffer cells of rats with fatty liver disease

<正>Objective:To observe the effects of different therapeutic methods and the recipes of Chinese medicine(CM) on the activation of c-Jun N-terminal kinase(JNK) in Kupffer cells of rats with fatty liver disease and to explore the mechanisms of these therapeutic methods.Methods:By using a random number table,98 rats were randomly divided into 7 groups:control group,model group,and 5 treatment groups,including soothing Liver(Gan) recipe group,invigorating Spleen(Pi) recipe group,dispelling dampness recipe group,promoting blood recipe group,and complex recipe group.Rats in the control group were fed with normal food and distilled water by gastric perfusion,while rats in the model group were fed with high-fat food and distilled spirits by gastric perfusion.Rats in the 5 treatment groups were fed with high-fat food and corresponding recipes by gastric perfusion.Twelve weeks later,all rats were sacrificed and liver tissues were stained for pathohistological observation.Kupffer cells were isolated from livers of rats to evaluate JNK and phospho-JNK expressions by Western blotting.Results:The grade of hepatic steatosis was higher in the model group than the control group(P0.05).Compared with the model group,the grade of fatty degeneration in soothing Liver recipe group and invigorating Spleen recipe group were significantly ameliorated(P0.05).Expressions of JNK and phospho-JNK in Kupffer cells were significantly higher in the model group than those in the control group(P0.05,P0.01).Compared with the model group,expressions of JNK in all treatment groups decreased,especially in invigorating Spleen recipe group and promoting blood recipe group(P0.05).Compared with the model group,expressions of phospho-JNK in all treatment groups declined significantly(P0.01),especially in soothing Live recipe group and invigorating Spleen recipe group. Conclusions:The high expressions of JNK and phospho-JNK in Kupffer cells might play an important role in the pathogenesis of fatty liver disease in rats.The recipes of CM,especially invigorating Spleen recipe and soothing Liver recipe,might protect liver against injury by reducing the total JNK protein content and inhibiting the activation of JNK protein in Kupffer cells of fatty liver model rats,which showed beneficial effects on fatty liver disease.     

Effects of Jinmaitong Capsule (筋脉通胶囊) on Ciliary Neurotrophic Factor in Sciatic Nerves of Diabetes Mellitus Rats

Objective:To study the effects of the Chinese medicine Jinmaitong Capsule(筋脉通胶囊,JMT) on the pathomorphology of sciatic nerves,ciliary neurotrophic factor(CNTF),and the mRNA expressions of CNTF in rats with streptozotocin-induced diabetes mellitus(STZ-DM).Methods:The animal model was established by one time intraperitoneal injection of streptozotocin.The rats were simply divided by random into 5 groups including model group,low-dose JMT group(JL),medium-dose JMT group(JM),high-dose JMT group(JH) and neurotropin group.For each of the above 5 groups,a group of 10 normal Wistar rats matched in body weight,age and gender were set as normal group.Intragastric administrations were started after the animal model established.The JL group were administered with five times the JMT dose recommended for a human adult;the JM group were administered with ten times the JMT dose recommended for a human adult;the JH group were administered with twenty times the JMT dose recommended for a human adult.The neurotropin group was administered with ten times the neurotropin dose recommended for a human adult.All rats were given intragastric administration for 16 weeks and then killed.In the 4th,8th,12th,16th week,body weight and blood glucose level were detected before and after the intervention.The morphologic changes of the sciatic nerves were observed by optical microscope and transmission electron microscope.The CNTFmRNA expressions were detected by real-time fluorescent quantitative polymerase chain protein,and the CNTF protein expressions were detected by immunohistochemical method.Results:The blood glucose levels of the STZ-DM rats were much higher than normal group(P<0.01),and there was no apparent difference between any treatment groups and the model group(P>0.05).Before and after the intervention in the 4th, 8th,12th,16th week,there were no significant differences in the body weight among all the groups(P>0.05). The sciatic nerves of STZ-DM rats might have pathomorphological changes in axons,myelin sheaths,and interstitium.The levels of CNTF and CNTF-mRNA expressions in the STZ-DM rats were both significantly decreased(P<0.01).The sciatic nerves of STZ-DM rats might have pathomorphological changes in axons, myelin sheaths,and interstitium.Conclusion:JMT could improve the pathomorphology of sciatic nerves by increasing CNTF’s and CNTF-mRNA expressions in sciatic nerve tissues,and promote the repair and regeneration of damaged nerve fibers.     

Effect of Qi-tonifying and Stasis-eliminating Therapy(益气活血法)on Expression of Vascular Endothelial Growth Factor and Its Receptors FIt-l,FIk-1 in the Brain of Intracerebral Hemorrhagic Rats

Objective:To investigate the effects and mechanism of qi-tonifying and stasis- eliminating(QTSE)therapy on the expression of vascular endothelial growth factor(VEGF)and its receptors Fit-1 and FIk-1 in the brains of intracerebral hemorrhagic(model)rats.Methods:One hundred and eighty Sprague-Dawley rats were randomly divided into six groups:the normal group (n=5),the sham-operative(SO)group(n=35),the model group(n=35),the QTSE group(n=35),the QT group(n=35)and the SE group(n=35).All the rats except those in the normal group and SO group were established into an intracerebral hemorrhage(ICH)model by intracerebral injection of collagenase typeⅦand the latter three were orally administered with Buyang Huanwu Decoction (补阳还五汤,a classical recipe for QTSE)or with some of its components for qi-tonification and for stasis-elimination,respectively.To the other three groups,normal saline solutions were given instead.Behavioral tests were carried out in the animals randomly chosen from each group on days 1, 2,4,7,14,21 and 28 after modeling.The expressions of VEGF,FIk-1 and Fit-1 were determined by immunohistochemistry and the number of vascular segments with positive expression in the injured brain area of the rats was calculated.Results:From day 7 onwards,the asymmetric forelimb use rate in the QTSE group recovered more significantly than that in the other model groups.In the model group,the expressions of VEGF,FIk-1 and Fit-1 appeared on day 1 and reached a peak on day 21, then weakened gradually.In the QTSE group,as compared with the other model groups,a higher level of VEGF expression was shown from day 7(P<0.01)and a higher level of Fit-1 expression was shown from the 7th day to the 21st day(P<0.01).Conclusion:QTSE therapy can up-regulate the expressions of VEGF and its receptors(FIk-1 and Fit-1)and improve the recovery of kinetic function in the ICH rats,which may be correlated with its action in modulating vascular regeneration to promote the reconstruction of microvascular networks in the damaged areas.     

Protective Effects of Traditional Tibetan Medicine Zuo-Mu-A Decoction (佐木阿汤) on the Blood Parameters and Myocardium of High Altitude Polycythemia Model Rats

Objective: To explore the protective effects of Tibetan medicine Zuo-Mu-A Decoction(佐木阿汤, ZMAD) on the blood parameters and myocardium of high altitude polycythemia(HAPC) model rats. Methods: Forty male Wistar rats were randomly divided into 4 groups by a random number table, including the normal, model, Rhodiola rosea L.(RRL) and ZMAD groups(10 in each group). Every group was raised in Lhasa to create a HAPC model except the normal group. After modeling, rats in the RRL and the ZMAD groups were administered intragastrically with RRL(20 mL/kg) and ZMAD(7.5 mL/kg) once a day for 2 months, respectively; for the normal and the model groups, 5 mL of distilled water was administered intragastrically instead of decoction. Then routine blood and hematologic rheology parameters were taken, levels of erythropoietin and 8-hydroxy-2'-deoxyguanosine(8-OHd G) were tested, and ultrastructural change in the left ventricular myocardium was observed using transmission electron microscopy. Results: Compared with the model group, ZMAD significantly reduced the red blood cell count, hemoglobin levels, whole blood viscosity at low/middle shear rates, plasma viscosity, erythrocyte electrophoretic time, erythropoietin and 8-OHd G levels, and also increased the erythrocyte deformation index(P0.05). There was no difference in all results between the RRL and the ZMAD groups. The cardiac muscle fibers were well-protected, mitochondrial matrix swelled mildly and ultrastructure changes were less prominent in the ZMAD group compared with the model group. Conclusion: ZMAD has significant protective effects on the blood parameters against HAPC, and also has the beneficial effect in protecting against myocardial injury.     

Effects of aerosolized ketamine on the level of nitric oxide and nitric oxide synthetase in the lung tissue of rat with asthma

Objective： To explore the effects of aerosolized ketamine on the level of nitric oxide and nitric oxide synthetase in the lung tissue in rat asthma model. Methods： Forty SD rats were randomly assigned to five groups： control group （group N）, asthma model group （group A）, two pretreated groups of different concentrations of ketamine （group K1, K2） and dexamethasone group（group D） with eight rats in each group. The rats in group A were sensitized by injection of ovalbumin （OA） together with aluminum hydroxide and bordetella pertussis as adjuvants. Two weeks after the sensitization, aerosolized OA was used to cause asthma. The rats in group K1 and K2 were sensitized with OA as group A , and then exposed to aerosol of ketamine , with the concentration of 25 g/L and 50 g/L respectively. Before using aerosolized OA,the rats in group D were exposed to aerosol of 0.01% dexamethasone .The level of NO2^-/NO3^- in lung tissues, inducible nitric oxide synthetase（iNOS） and constitute nitric oxide synthetase（cNOS） was measured in all groups. Results： The level of NO2^-/NO3^- and the activity of iNOS in lung tissues in group A were signiticantly higher than those in the other groups. The iNOS activity and the level of NO2^-/NO3 ^- in lung tissues were highly positively correlated. Conclusion： NO can induce airway hyperreactivity that may worsen asthma. Aerosolized ketamine can decrease the iNOS expression and reduce the level of NO in the lung tissue in rat asthma model.     

Effect of Qingre Buyi Decoction (清热补益汤) on Nitric Oxide and Histomorphology of Intestinal Mucosa in Rats with Radiation Enteritis

Objective: To explore the mechanism of Qingre Buyi Decoction (清热补益汤,QBD) in prevention and treatment of radiation enteritis in rats. Methods: Forty-eight Wistar rats were randomly divided into four groups, the TCM group, the WM group, the model group and the control group, 12 in each group. Rats in the former three groups were given orally with QBD, norfloxacin and normal saline once a day for 7 successive days, after being irradiated with X-ray at single dose of 10 Gy for modeling of radiation enteritis, while rats in the control group were untreated. Animals were sacrificed at the end of the medication. NO concentration, mean height and number of villi per centimeter in their small intestinal mu-cosa were measured. Results: The intestinal NO concentration was significantly lower in the TCM and WM groups than that in the model group(P<0.05), while the number of villi was significantly more and the height higher in the former two groups than those in the model group (P<0.01 for both), but no signif     

Chinese Herbal Formula Ermiao Powder (二妙散) Regulates Cholinergic Anti-inflammatory Pathway in Rats with Rheumatoid Arthritis

Objective: To investigate the effect of Chinese herbal formula Ermiao Powder (二妙散, EMP) on the expression of cholinergic anti-inflammatory pathway in rats with rheumatoid arthritis (RA). Methods: Seventy two rats were randomly ivided into 6 groups according to body weight, including normal control group, collagen induced arthritis (CIA) group, three doses EMP groups, and methotrexate (MTX) group (n=12 per group). All of the rats except for those in the normal control group were given multipoint subcutaneous injection of bovine type Ⅱ collagen to establish a CIA model. Three EMP groups received a high- (4.5 g/kg), medium- (3.0 g/kg), and low- (1.5 g/kg) doses of EMP by intragavage, respectively. MTX group was injected intraperitoneally MTX at 0.9 mg/kg once a week as the positive control. The administration was 3 consecutive weeks. Joint swelling, arthritis index, and body weight changes in different experimental groups of rats were tested. The joint damage was evaluated by masson staining. Quantitative real-time polymerase chain reaction, Western blot, and immunohistochemistry (IHC) were performed to evaluate the expression of CHRNA7, encoding α 7 nicotinic acetylcholine receptor in the cholinergic anti-inflammatory pathway, in different tissues and their localization in the spleen and joints. Results: CHRNA7 expression levels were significantly higher in the joints and spleens of CIA group than those in normal control group (both P<0.05). Moreover, the CHRNA7 mRNA and protein levels in the spleen and joints of MTX and three doses of EMP groups were significantly lower than CIA group (all P<0.05). Compared with the MTX group, treatment with low-dose EMP resulted in significant reduction of CHRNA7 mRNA and protein expression levels (P<0.05 or P<0.01). IHC showed positive signals of CHRNA7 in the white pulp and red pulp of the spleens of rats; CHRNA7 was expressed on fibroblast-like synoviocytes, macrophages, and endothelial cells in the joints of rats, and the expression in the joints of low-dose EMP group was significantly lower than that in the CIA group (P<0.01). Conclusion: Cholinergic anti-inflammatory pathway was involved in the generation of the inflammatory reaction in CIA rats, and EMP exerted therapeutic effect on RA through cholinergic anti-inflammatory pathway.     

Effects of Yinchenhao Decoction(茵陈蒿汤) for Non-alcoholic Steatohepatitis in Rats and Study of the Mechanism

Objective:To observe the effects of Yinchenhao Decoction(茵陈蒿汤) for non-alcoholic steatohepatitis(NASH) in rats and study the mechanism.Methods:Total 18 male SD rats were randomly divided into a normal control group,a model group and a treatment group,6 rats in each group.Rats in the model and treatment groups were fed with high-fat forage for 10 weeks to prepare the NASH model,and the rats in the treatment group were administrated with Yinchenhao Decoction from the 6th week for 5 weeks.All rats were sacrificed at the end of the10th week and the samples were collected.Serum alanine aminotransferase(ALT) activity,tumor necrosis factor-α(TNF-α) level,and hepatic triglyceride(TG) and free fatty acid(FFA) contents were determined.Hepatic pathological changes were detected by HE staining.Results:Serum ALT activity,TNF-α level,hepatic TG and FFA contents,and the fatty deposition in hepatocytes were significantly reduced in the rats of the treatment group.Conclusion:Yinchenhao Decoction has good therapeutic effects for NASH,protecting the liver function and reducing the fatty deposition in liver,which are possibly related with reduction of FFA content and inhibition of TNF-α expression.     

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.     

祛痹镇痛方对胶原蛋白诱导的关节炎大鼠滑液基因表达的作用

Objective: To investigate the effect of the Chinese medical formula Qubi Zhentong Recipe (祛痹镇痛方, QZR) on the synovial gene expression pro?le in collagen-induced arthritis (CIA) rats. Methods: Ten rats were randomly chosen from 60 rats as the control group, and the other 50 rats were used for the CIA models. The CIA model group was constructed by bovine injection of type Ⅱ collagen through the rats'' neck and tail. Twenty rats were randomly chosen from 34 successful CIA models and randomly assigned into two groups: the model group (n=10) and the QZR group (n=10). The QZR group was fed intragastrically with QZR 22.9 g/(kg?d) (10 times the clinical adult dose), and the CIA model group was given the same dose of normal saline. Both model and QZR groups were administered treatment once a day. Total RNA was collected from the knee joint synovium after 30 days. The change in gene expression pro?le was analyzed by a whole gene chip. Results: A total of 76 genes showed a difference in expression between CIA model group and the control group; 35 genes were down-regulated and 41 were up-regulated. A total of 67 genes showed a difference in expression between the model group and the QZR group; 48 genes were down-regulated and 19 were up-regulated. Conclusions: QZR may affect CIA by stimulating multiple genes and targets, which are related to oncogenes, apoptosis, metabolism, the immune system, ion channels, and transport proteins.     

针灸脑缺血再灌注损伤大鼠百会及足三里穴位对热休克蛋白70及肿瘤坏死因子-α表达的作用

Objective:To explore the effects of acupuncture on the peripheral serum expression of heat shock protein 70(HSP70)and tumor necrosis factor a(TNF-α)in rats with cerebral ischemia-reperfusion injury(CIRI).Methods:In total,152 Sprague-Dawley(SD)rats were randomly divided into an operated group and a non-operated group according to a random digits table.The operated group included a sham-operated group,a model group and an acupuncture group,whereas the non-operated group consisted of a normal group.Except for the normal group,each group was further divided into 12,24,48,72,96,and 144 h time points according to different reperfusion times.Eight rats were assigned in each operated group and in the normal group.The rat model of CIRI was established by the thread occlusion method in the model and acupuncture groups.The acupuncture group was treated with electroacupuncture at Baihui(DU20)and Zusanli(ST36)for the required time after successful operation.Blood was sampled to detect the HSP70 and TNF-αcontent by enzyme linked immunosorbent assay.Results:The expression of HSP70 protein in the peripheral serum of the experimental groups was higher than that in the normal control group.The peak time in both the model and the sham-operated groups was 12 h,and the peak time in the acupuncture group was 24 h.The expression in the acupuncture group declined to a lower level at 72 h and was lower than that in the model and sham-operated groups(P0.05).The peak time for the expression of TNF-αprotein in the peripheral serum of both the model and the acupuncture groups was 24 h,but the expression in the acupuncture group was lower than the model group.Additionally,the expression of TNF-αin all experimental groups was higher than the normal group(P0.05).Conclusions:Acupuncture at DU20 and ST36 in rats attenuated CIRI,which was associated with a reduction in the expression of HSP70 and TNF-α.These results provide clues to acupunctural neuroprotective properties.Acupuncture at DU20 and ST36 in rats after CIRI can adjust the expression of HSP70 and TNF-αin the peripheral serum,which might be one of the mechanisms of acupuncture's attenuation of CIRI.     

Effects of ω-3 polyunsaturated fatty acids on prostaglandin E-2 and arachidonic acid in rats with senile dementia

Objective To investigate the mechanism of ω-3 polyunsaturated fatty acids in senile dementi-a. Methods The normal 15 months old Wistar rats were randomly divided into normal control,model group andre-cape latency time and mistaken times were evaluated by using the step down test. The content of serum PGE2 and urachidonic acid(AA),EPA and DHA of cerebral tissue in rats were detected. Results The lingeringly latency and error times were ( 264.83±16.99) s and (4.8±1.7) times in control group, ( 189.26±31.42)s and ( 9.6± 2.2) times in model group, (230.88±29.35) s and (7.3±2.2) times in treatment group, respectively. Differences in three groups were very significantly by analyses of variance(F=20.114, F=13.638, P=0.000). Compared with model group, control group and treatment group had lingeringly latency(LSD-t=6.332,P=0.000 and LSD-t =3.987, P=0.002, respectively) and less error times (LSD-t=5.221, P=0.000 and LSD-t=3.502, P= 0.019,respectively). Serum PGE2 were in control group, model group and treatment group were very significantly by analyses of variance(F=6.851 , P=0.004). Compared with model group,control group had less serum PGE2 (LSD-t=3.684, P=0.001 ). Compared with model group,ω-3 polyunsaturated fatty acids chould decrease serum PGE2( LSD-t=2.152, P=0.041 ). AA, EPA and DHA of cerebral tissue in control group, in model group, in treatment group, respectively. Differences in three groups were very significantly by analyses of variance (F=5.538, P=0.010, F=4.240, P=0.025, F=4.633, P=0.019). Compared with model group,ω-3 polyunsaturat-ed fatty acids could decrease A A(LSD-t=2.273, P=0.031) and increase EPA and D HA (LSD-t=2.428, P= 0.022,LSD-t=2.520, P=0.018,respectively). Conclusions ω-3 polyunsaturated fatty acids can improve learning-memory function in rats,change the proportion of AA,EPA and DHA in cerebral tissue and prevent senile dementia.     

Structural Modulation of Gut Microbiota in Rats with Allergic Bronchial Asthma Treated with Recuperating Lung Decoction

ObjectiveTo investigate whether recuperating lung decoction (RLD) can modulate the composition of gut microbiota in rats during asthma treatment. MethodsFifteen Sprague-Dawley rats were divided randomly and equally into control group, model group, dexamethasone (DEX) group, RLD medium-dose group, and RLD high-dose group. The asthma model was established in all groups, except for the control group. The rats in the DEX and RLD groups were treated orally with DEX and RLD, respectively. The rats in the control and model groups were treated orally with 0.9% saline. The intestinal bacterial communities were compared among groups using 16S rRNA gene amplification and 454 pyrosequencing. ResultsThe microbial flora differed between the control and model groups, but the flora in the RLD groups was similar to that in the control group. No significant differences were observed between the RLD high-dose and medium-dose groups. RLD treatment resulted in an increase in the level beneficial bacteria in the gut, such asLactobacillusandBifidobacteriumspp. ConclusionOral administration of RLD increased the number of intestinal lactic acid-producing bacteria, such as Lactobacillus andBifidobacterium, in asthma model rats.     

曾耀明 郑伟伟 余维微 陈余圣 林上助

Objective: To observe the level of serum CGRP and 5-hydroxytryptamine in animals model of irritable bowel syndrome treated with sheng yang yi wei decoction.Methods: 70 SPF SD rats were divided randomly into normal control group,model group,dicetel group,sheng yang yi wei decoction group(low dosage,moderate dosage,high dosage).The rat model of irritable bowel syndrome was made by binding stress and clasping their tails.group.Changes of diarrhea rate were observed in these groups．Radioimmunoassay was used to detect the level of CGRP and 5-HT in plasma．Results：The diarrhea rates in group were decreased after treatment,and had the statistics significance in comparison with model group and dicetel group(P<0.05)．The contents of 5-HT in plasma in model group were higher than those in normal control group,while the levels of CGRP in model group were slighly lower than those in normal control group(P<0.05).After treatment,the contents of 5-HT in plasma in dicetel group and sheng yang yi wei decoction group were markedly decreased than those in model group(P<0.05),while the levels of CGRP had no statistics significance among all groups(P>0.05).Conclusions: Sheng yang yi wei decoction can markly alleviate the symptom of diarrhea,inhibit 5-HT synthesis and secretion in animal model of irritable bowel syndrome,while it does not effect the levels of CGRP.