Effects of active components of Red Paeonia and Rhizoma chuanxiong on angiogenesis in atherosclerosis plaque in rabbits

Objective： To investigate the effects and mechanism of the active components of Red Paeonia and Fthizoma chuanxiong （Xiongshao Capsule, 芎芍胶囊, XSC） on angiogenesis in atherosclerosis plaque in rabbits. Metbods： Fifty New Zealand rabbits were randomly divided into the normal group, the model group, and the three medicated groups treated respectively with Simvastatin （2.5 mg/kg per day）, low-dose （0.24 g/kg per day） and high-dose （0.48 g/kg per day） XSC, 10 in each group. Rabbits in the normal group were fed with regular diet. To those in the other four groups, high fat diet was given, and a balloon angioplasty was performed two weeks later to establish abdominal aortic atherosclerosis model. Then, the model rabbits were fed continuously with high fat diet, and to those in the medicated groups, the testing drugs were added in the forage correspondingly for 6 successive weeks. Levels of blood lipids were measured at the end of the experiment. Meantime, serum levels of high sensitivity C-reactive protein （hsCRP） and tumor necrosis factor α （TNF-α） were detected with enzyme-linked immunoassay; the plaque area （PA）, cross-sectional vascular area （CVA） and correcting plaque area （PNCVA） were determined quantitatively using imaging software; and the protein expression of vascular endothelial growth factor （VEGF） and factor Ⅷ related antigen （FⅧRAg） in plaque was detected using immunohistochemical method. Results： As compared with the model group, the content of total cholesterol （TC） in the three medicated groups, and contents of triglyceride （TG） and low-density lipoprotein cholesterol （LDL-C） in the Simvastatin group were lower to various extents （P〈0.05, P〈0.01）. The serum level of hsCRP in all modeled rabbits was higher than that in the normal group, but in the three treated groups it was significantly lower than that in the model group （P〈0.05, P〈0.01）. Expressions of VEGF and FⅧRAg, as well as PNCVA in the three medicated groups were significantly lower than those in the model control group （P〈0.05, P〈0.01）. Conclusion： The active components of Red Paeonia and Rhizoma chuanxiong have definite effects in delaying the genesis and development of atherosclerosis, its mechanism might be related with the inhibition on angiogenesis in plaque, and also with its actions of lipo-metabolism regulation and anti-inflammation.     

Effects of Xianzhong Injection on the Function of Knee Joint and the Content of Insulin-Like Growth Factor-1 in Rabbits of Early Osteoarthritis

Objective: To evaluate the effect of Xianzhong Injection (仙仲注射液) into the articular cavity on the function of the joints and the content of insulin-like growth factor-1(IGF-1) in rabbits with early knee osteoarthritis (OA). Methods: 120 rabbits were randomly divided into five groups: blank group, model group (injecting 0.5ml of normal saline, once a week), positive control group (injecting 0.5ml of sodium hyaluronate, once a week), Xianzhong group I (injecting 0.5ml of Xianzhong Injection, once a week), Xianzhong group II (injecting 0.5ml of Xianzhong Injection, twice a week). Changes of the articular function and IGF-1 levels in the serum and joint fluid were investigated 8 weeks later. Results: The function of knee joint in the Xianzhong group I and II was improved significantly as compared with that in the model group (P<0.01), and there was no significant difference between the Xianzhong group I and the positive control group (P>0.05), but a significant difference was seen between the Xianzhong group I and the Xianzhong group II (P<0.05); in the positive control group, the Xianzhong group I and the Xianzhong group II , the levels of IGF-1 in the joint fluid and serum were significantly increased as compared with that in the model group (P﹤0.01). Conclusion: Xianzhong Injection can improve the function of knee joint in rabbits with early knee OA, and can substitute sodium hyaluronate Injection for treating this disease.     

Experimental Study on Gufusheng in Treatment of Steroid-Induced Ischemic Necrosis of Femoral Head in Rabbits

Objective： To explore.the mechanisim of compound Gufusheng in treatment of steroid-induced ischemic necrosis of femoral head （SINFH）. Thirty-two rabbits were randomly divided into a blank control group （n=12） and a model group （n=20）. The rabbits of the model group were established by intramuscular injection of prednisone acetate （0.32mg/kg/d） for 8 weeks, and 2 rabbits in each group were killed at the 6th and the 8th week respectively to investigate whether the model was successfully prepared. Then remainder in the model group was again randomly divided into group A （Gufusheng group, n=8） and group B （model group, n=8）, while the rest of rabbits in the blank group constituted group C （n=8）. The rabbits in group A were given Gufusheng Decoction （骨复生煎液） by intragastric perfusion, and the rabbits in group B and C were fed with normal saline. After treatment for 4 weeks, the serum insulin-like growth factor-1 （IGF-1） and tumor necrosis factor-~ （TNF-α） levels in the rabbits of the 3 groups were determined by radioimmunoassay （RIA）. The serum TNF-α decreased significantly and the IGF-1 level increased significantly in the Gufusheng group as compared with the model group （both P〈0.01）. Compound Gufusheng can prevent SINFH and promote repair of the tissue of necrosis of femoral head possibly via decreasing the TNF-α and increasing IGF-1 level.     

改良补肾活血方对实验性骨关节炎兔子模型的控制代谢及抗氧化作用

Objective: To observe the metabolic, regulatory and anti-oxidative effects of modified Bushen Huoxue Decoction (BSHXD, 补肾活血方), a Chinese herbal medicine for Kidney (Shen)-reinforcement and blood-activation, on an osteoarthritis (OA) rabbit model. Methods: A rabbit model for knee joint OA was established by the classic Hulth''s method. The OA model rabbits were randomized into 5 groups: the model control group, the positive control group treated with glucosamine sulfate, and the three BSHXD treated groups treated respectively with low, moderate, and high doses of BSHXD. In addition, a normal control group and a sham-operated group were set up. Experimental animals were sacrificed after a 7-week treatment, and pathological changes in cartilaginous tissue were estimated using the Mankin criteria. Hydroxyproline (Hyp) and malonaldehyde (MDA) contents in blood serum and urine, as well as superoxide dismutase (SOD) activity and nitric oxide (NO) content in blood serum and knee joint synovial homogenates were detected. Results: Mankin scoring showed insignificant statistical differences between the various treatment groups (P>0.05), but all were better than the model control group (P<0.05). Serum and urinary contents of Hyp and MDA as well as serum and synovial levels of NO were significantly lower, but the SOD activity in blood serum and synovial tissue was higher in the BSHXD treated groups than in the model group (P<0.01); the effect of BSHXD was dose-dependent to some extent. Conclusion: The modified BSHXD shows an effect of improving cartilage metabolism in experimental rabbits with OA, and possesses osteo-chondric protective effects in antagonizing peroxidation injury.     

Effects of Simvastatin on NF-kB-DNA Binding Activity and Monocyte Chemoattractant Protein-1 Expression in a Rabbit Model of Atherosclerosis

To observe the effects of simvastatin on nuclear factor kappaB （NF-kB）-DNA binding activity and on the expression of monocyte chemoattractant protein-1 （MCP-1） in atherosclerotic plaque in rabbits and to explore the anti-atherosclerotic properties beyond its lipid-lowering effects. Thirty-six New Zealand male rabbits were randomly divided into low-cholesterol group （LC）, high- cholesterol group （HC）, high-cholesterol＋ simvastatin group （HC＋S） and then were fed for 12 weeks. At the end of the experiment, standard enzymatic assays, electrophoretic mobility shift as- say （EMSA）, immunohistochemical staining, and morphometry were performed to observe serum lipids, NF-kB-DNA binding activity, MCP-1 protein expression, intirna thickness and plaque area of aorta respectively in all three groups. Our results showed that the serum lipids, NF-kB-DNA binding activity, expression of MCP-1 protein, intima thickness, and plaque area of aorta in the LC and HC＋S groups were significantly lower than those in the HC group （P〈0.05）. There was no significant difference in the serum lipids between the LC and HC＋S groups （P〉0.05）, but the NF-kB-DNA binding activity, the expression of MCP-1 protein and the intirna thickness and plaque area of aorta in the HC＋S group were significantly decreased as compared to the LC group （P〈0. 05）. This study demonstrated that simvastatin could decrease atherosclerosis by inhibiting the NF-kB-DNA binding activity and by reducing the expression of MCP-1 protein.     

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.     

Preventive Administration of Juanbi Capsules (蠲痹胶囊) for Knee Osteoarthritis: Effects on Serum MMP-2 and MMP-9 levels and Cartilage Repair

Objective: To explore the mechanism of action of Juanbi Capsules (蠲痹胶囊), a Chinese medicine for invigorating the kidney and replenishing qi, in preventing osteoarthritis of the knee in rabbits. Methods: Seventy-two 4-month-old, Japanese long-eared white rabbits were randomly divided into 6 groups: control (group A), model (group B), Chinese drug; high-dose (group C), Chinese drug; mid-dose (group D), Chinese drug; low-dose (group E), and drug control (group F). With the exception of the rabbits in group A, each rabbit was subjected to plaster cast fixation for 6 weeks to induce osteoarthritis. In addition, rabbits were administrated with an intragastric injection of the Chinese drug (groups C, D and E) or an aminoglucose hydrochloride capsule (group F) for 4 weeks. Blood was drawn from the central ear artery for serum MMP-2 and MMP-9 concentrations, and the knee joint cartilage was harvested for gross observation and light microscopy. Results: There were significant differences in serum MMP-2 and MMP-9 concentrations between group B and groups C, D and E (P<0.05), with no significant differences between groups D and F. Histological results showed various changes in tissue staining with treatment, with osteophyte and bone cyst formation, and superficial erosion in the articular surface of the cartilage; in some cases, the defect reached the mid-layer of the cartilage, and these changes were lower than those in the model group. Conclusion: Juanbi Capsules assist in preventing osteoarthritis in the rabbit, possibly by decreasing serum MMP-2 and MMP-9 levels.     

Effects of simvastatin on early oxidative stress and endothelial function in apolipoprotein E-deficient mice

Objective:To investigate the mechanisms that Simvastatin,a 3-ydroxy-3-methylglutaryl coenzyme A(HMG-CoA)reductase inhibitor,plays an important role in primary prevention of atherosclerosis independently of its lipid-lowering effect in Apolipoprotein E-deficient mice in the early stage of atherosclerosis.Methods:Twenty-four 6-week old male apoE-deficient mice were randomly divided into two groups:control group(normal saline)and treatment group[simvastatin(5 mg/(kg·d))].Simvastatin was administered to treatment group mice by gavage and the same volume of normal saline was administered to control group mice by the same method for 2 or 4 weeks.Total cholesterol(TC),super-oxide dismutase(SOD),malondialdehyde(MDA)and serum nitric oxide(NO)were measured by biochemical analysis.Results:There was no significant difference in serum TC between control and treatment groups.Compared with the control's,the effects of simvastatin were more signiticant in decreasing serum MDA level(P＜0.01 vs control's at 2-week;P＜0.006 vs control's at 4-week),increasing serum SOD level(P＜0.03 vs control's at 2-week;P＜0.003 vs control's at 4-week)and NO level (P＜0.01 control's at 2-week;P＜0.001 vs control's at 4-week)either at 2 or 4 weeks.Conclusion:Simvastatin attenuates oxidative stress and protects endothelial function by the mechanisms of decreasing serum MDA level,increasing serum SOD level and NO level,which were inconsistent with its cholesterol-lowering effect.It may play an important role in primary(if not all)prevention of atherosclerosis and might be independent of lipid-regulation mechanism.     

Experimental study of changes of TNF-α in rabbits with steroid-induced avascular necrosis of femoral head

Objective: To explore the effect of tumor necrosis factor-α(TNF-α)on the occurrence of steroid-induced avascular necrosis of femoral head(SANFH). Methods: Twenty-four rabbits were firstly divided into void group ( n = 12) and model group ( n =two groups were killed respectively to observe whether the model was successful. The level of TNF-α in serum of the residual rabbits of the two groups was examined in Radioimmunoassay method. Results: The level of TNF-α in model group is significantly higher than that in void group( P ＜0.001) under the premise of the model of SANFH success by histological observation. Conclusion: The rise of level of TNF-α may be one of the most important factors in the occurrence of SANFH.     

Effects on the Tumor Specific Growth Factor and Tumor Necrosis Factor α in Rats' Precancerous Lesion of Primary Hepatocellular Carcinoma by Direct Moxibustion at Ganshu(BL 18) Acupoint

Objective: To investigate the effect of direct moxibustion at Ganshu(BL18) on the serum concentrations of tumor specific growth factor(TSGF) and tumor necrosis factor α(TNF-α) in a rat model with precancerous lesion of primary hepatocellular carcinoma(HCC), so as to explore the mechanism of moxibustion underlying improvement of HCC. Methods: Sixty male Wistar rats were randomly divided into control group(n=10), model group(n=20), prevention group 1(n=15) and prevention group 2(n=15). The normal rats were injected with physiological saline as blank control. At the same time, the rats of other three groups were injected with diethylnitrosamine to establish the HCC model. Direct moxibustion with grain-sized moxa was applied to bilateral Ganshu acupoint of the rats in the prevention group 1(1 treatment course, 20 days) and prevention group 2(2 treatment courses, 40 days), 5 doses for each acupoint, 0.5 mg/dose, once every other day. At each time point(before model establishment, the end of 1st course prevention, the end of 2nd course prevention and the end of model establishment), serum levels of TSGF and TNF-α were detected using enzyme-linked immunosorbent assay. Results: Compared with the control group, there was a remarkably increase of serum TSGF and TNF-α contents in the model group at the end of the experiment(P0.05). At the end of the 1st course of direct moxibustion, the contents of serum TSGF and TNF-α of rats in the prevention group 1 were significantly increased compared with that of the model group(P0.05). At the end of the 2nd course of direct moxibustion, serum TSGF and TNF-α levels of rats in the model group were higher than the normal group with significantly difference(P0.05), and the levels of TSGF and TNF-α in the prevention group 2 were significantly reduced in comparison with the model group(P0.05). Conclusion: It was possible that direct moxibustion could inhibit precancerous lesion and postpone hepatocarcinogenesis, and the therapeutic effect of two courses were better than one course.     

Effect of Xiongshao Capsule（芎芍胶囊）on the Function of Vascular Endothelium of Patients with Cervical Atherosclerosis

Objective: To study the effect of Xiongshao Capsule (芎芍胶囊, XSC), a TCM herb that can promote blood circulation to remove blood stasis, on the endothelial dependent relaxation function, serum nitric oxide (NO), and plasma endothelin-1 (ET-1) of the patients with cervical atherosclerosis. Methods: Forty patients were randomly divided into two groups: XSC group and Probucol group (western medicine control).In addition, 20 healthy people were set as a normal control group. Plasma ET-1, serum NO, the internal diameter of basal brachial artery, endothelial dependent flow mediated dilation (FMD) and non-endothelial dependent nitroglycerin induced dilation (NID) to the trial group before and after therapy and to the healthy control group were determined respectively. Results: Compared to the healthy control group, FMD of patients with atherosclerosis was damaged obviously, the serum NO level decreased, plasma ET-1 increased (P＜0.01), NID also decreased (P＜0.05), the internal diameter of basal brachial artery has no obvious difference (P＞0.05). After the patients with atherosclerosis were treated with Xiongshao Capsule for 12 weeks,FMD increased evidently, plasma ET-1 decreased, serum NO and the ratio of NO/ET-1 increased, compared with the level before therapy and Probucol group, the difference was significant (P＜0.05, P＜0.01), NIDdidn′t change obviously (P＞0.05). Conclusion: XSC could regulate vascular activity factor and improve the function of endothelial dependent vascular dilation of patients with atherosclerosis.     

Effect of Chengzai Pill on the L-Type Ca^2＋ Channel Currents of Osteoblasts Pretreated with Methylprednisolone

Objective: To observe the effect of the serum containing Chengzai Pill on the L-type voltage-sensitive calcium channels current (L-VSCCsC) of osteoblastic MC3T3-E1 cells pretreated with methylprednisolone (mPSL). Methods: A control group, a model group, a low dose group and a high dose group were set up. The whole cell patch clamp technique was used to record L-VSCCsC of 10 osteoblastic MC3T3-E1 cells in each group and their peak currents were determined. Results: The peak current of the control group was 0.2284±0.0209 nA; the peak current of the model group was 0.1839±0.0179 nA; decreased by 19.5% as compared with the control group (P<0.01); the peak current of the low and high dose groups was 0.2526± 0.0093 nA and 0.2671±0.0120 nA respectively, increased by 37.4% and 45.2% as compared with the model group (P<0.01); the difference between the low and high dose groups was P<0.05. Conclusion: 1. mPSL inhibits L-VSCCsC of osteoblasts; and 2. The serum containing Chengzai Pill increases L-VSCCsC of osteoblasts pretreated with mPSL.     

Effects of Bushen Huoxue Decoction on Nitric Oxide （NO） in Serum, Articular Cartilage and Synovium in Rabbits of Knee Osteoarthritis

Forty-eight New Zealand rabbits were divided into normal group(n=18),control group(n=18)andChinese herbs treatment group(n=12)randomly.The rabbits in the normal group receivedsham-operation,and the OA model was established by Hulth's method.All the rabbits in the treatmentgroup were given Bushen Huoxue Decoction(补肾活血汤)from the 6th week after the operation.At 6th,8th and 12th week after the operation,the NO concentrations of the serum,joint cartilage and synoviumwere examined.Results:Indicated that the NO concentrations of the serum,joint cartilage and synoviumin the control group were all significantly higher than those in the normal group,with the joint cartilagemore obvious(P<0.05).In the Chinese herbs treatment group the NO concentrations in all the partsobviously decreased as compared with the control group(P<0.05).It is suggested that Bushen HuoxueDecoction decrease the levels of NO in the serum,synovium and joint cartilage in the OA rabbit.     

Effect of Compound Laser Acupuncture-Moxibustion on Blood Glucose, Fasting Insulin and Blood Lipids Levels in Type 2 Diabetic Rats

Objective: To investigate the effect of compound laser acupuncture-moxibustion on blood glucose, fasting insulin and blood lipids levels in type 2 diabetes mellitus(T2DM) rats. Methods: Forty male Wistar rats were randomly divided into 4 groups, including the normal group, model control group, laser group and sham laser group(n=10 per group). The rats in the normal group were fed with a standard diet. Rats in other groups were fed with a high-sugar and high-fat diet for 4 weeks, then intraperitoneally injected with 1% streptozotocin to induce T2 DM model. The laser group was irradiated by 10.6 μm and 650 nm compound laser on bilateral Pishu(BL 20), Shenshu(BL 23) and Sanyinjiao(SP 6) for 5 min, 6 times a week for 5 weeks. The sham laser group received the same treatment as the laser group, but without laser output. The model control group and normal group were not treated. Blood glucose levels were measured before and after 1, 2, 3, 4 and 5 weeks of treatment. The serum levels of fasting insulin, total cholesterol(TC), triglyceride(TG), low-density lipoprotein(LDL), and high-density lipoprotein(HDL) were analyzed after the last treatment. Results: The blood glucose levels in the model control group increased during the 5 weeks of treatment compared with the normal group(P0.05), while those in the laser group were significantly lower than the model control group after weekly treatment(P0.01 or P0.05). After 1, 2 and 3 weeks of treatment, the blood glucose levels in the laser group decreased obviously compared with the sham laser group(P0.01 or P0.05). Compared with the normal group, the levels of fasting insulin, TC and LDL in the model control group notably increased(P0.01 or P0.05), while their levels in the laser group were significantly lower than the model control group after 5 weeks of treatment(P0.05 or P0.01). However, no statistically significant differences were observed in TG or HDL levels among the 4 groups(P0.05). Conclusion: The compound laser acupuncture-moxibustion of 10.6 μm and 650 nm had positive effects on the regulation of hyperglycemia and insulin resistance in T2 DM rats, which may be a potential treatment for T2 DM, and also provide an alternative to the traditional acupuncture and moxibustion therapy.     

Effects of bone morphogenetic protein on the osseointegration of porous-surfaced implants：An experimental study in rabbits

Objective： Research on enhancing early osseointegration of cementless implants to improve early fixation and reducing of risk of loosening. Methods ： Thirty New Zealand rabbits were divided into two groups at random. BMP combined with DBM 30 mg was inserted around the prosthesis in 15 rabbits as experimental group, the remaining rabbits were served as control group. After 4, 8, and 12 weeks, five rabbits were sacrificed in each group. The humerus with the implants were retrieved. Bone ingrowth was analyzed by none-decalcification bone ground section and biomechanical test. Results： At the end of 4 and 8 weeks the osseointegration rates of BMP group were higher than those of control （P 〈 0.05）. The ultimate shear strength between BMP treated implantation and the control was the same as the results in osseointegration rates at 4, 8 weeks （P 〈 0.05）. However, there was no difference between the treated and untreated group in the osseointegration rate and ultimate shear strength at 12 weeks （P 〉 0.05）. Conclusion： BMP combination can enhance bone growth into gaps around cementless implants, especially in the early postoperative period.     

Assessing the validity of a novel model of vertebral artery type of cervical syndrome induced by injecting sclerosing agent next to transverse process of cervical vertebra

The efficacy of injecting sclerosing agent next to transverse process of cervical vertebra to induce vertebral artery type of cervical syndrome(CSA)was observed.Twenty rabbits were randomly divided into two groups:the model group and the control group.The rabbits in the model group were injected with sclerosing agent next to transverse process of cervical vertebray,on the contrary,the rabbits in the control group were injected with nothing.Transcranial Doppler(TCD)was used to detect the average speed of blo...     

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.     

Effect of chinese herbal drug-containing serum for activating-blood and dispelling-toxin on ox-LDL-induced inflammatory factors’ expression in endothelial cells

<正>Objective:To investigate the effects of drug-containing serum of Chinese herbal compound, Xiongshao Capsule(芎芍胶囊,XS,for activating-blood) and Huanglian Capsule(黄连胶囊,HL,for dispellingtoxin) on the oxidized low-density lipoprotein(ox-LDL)-induced inflammatory factors in human umbilical vein endothelial cells(HUVECs).Methods:Thirty-two rats were randomly divided into four groups:the blank control group treated with distilled water,the positive control group treated with simvastatin(1.8 mg/kg),the test group I treated with Chinese herbal compound of XS(0.135 g/kg),and the test group II treated with Chinese herbal compound of XS(0.135 g/kg) and HL(0.135 g/kg).All the treatments were administered for 7 successive days by gastrogavage.Rats' blood serum was harvested 1 h after the last administration to prepare respective drugcontaining serum.HUVECs were exposed to ox-LDL(100μg/mL) to induce cell injury model and incubated with corresponding drug-containing serum for 24 h.Untreated HUVECs were set for blank control.Levels of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),and soluble intercellular adhesion molecule-1(slCAM-1) in supernatant of cultured HUVECs were determined by enzyme-linked immunosorbent assay(ELISA).HUVEC surface expressions of ICAM-1 and E-selectin were determined by flow cytometry.Results:Levels of IL-6, TNF-α,and slCAM-1 in the supernatant of HUVECs as well as the cell surface expressions of ICAM-1 and E-selectin significantly increased after 24-h ox-LDL stimulation(P0.01),while the abnormal elevations,except slCAM-1 in the test groupⅠ,were all reduced in the treated groups(the positive control and the two test groups) significantly(P0.01 or P0.05).Besides,the effect in the test group II seemed somewhat higher than that in the test group I but with no statistical significance(P0.05).Conclusion:Drug-containing serum of XS plus HL has a certain inhibitory effect on the vascular endothelial inflammation response induced by ox-LDL.     

Effects of Bushen Tiaochong Recipe (补肾调冲方) containing serum on ovarian granulosa cell proliferation, steroidogenesis and associated gene expression in rats

Objective:To observe the effect of Bushen Tiaochong Recipe (补肾调冲方,BSTCR) on rats' ovarian granulosa cell (GC) proliferation,steroidogenesis and follicle-stimulating hormone receptor (FSHR),and insulin-like growth factor-1 (IGF-1) mRNA expression using serum pharmacological method.Methods:Rats' GCs were incubated with 10% blank serum (as negative control group),follicle- stimulating hormone (FSH)-containing serum (S-FSH,as positive control group),or BSTCR (in different dosages) containing serum (S-BSTCR,as the BSTCR groups) for 48 h.~3H-TdR incorporation was then performed;DNA was measured to analyze the distribution of GCs in the cell cycle and their proliferation index (PI) using a flow cytometer;estradiol (E_2) and progesterone (P) content in the culture fluid were examined by radioimmunoassay;and levels of FSHR and IGF-1 mRNA expression in GCs were measured by real-time RT-PCR.Results:A dose-dependent increase of ~3H-TdR incorporation in GC was shown in the BSTCR groups.Cells in G_0/G_1 phase had markedly less,while those in S phase had a significantly higher increase in the BSTCR groups compared with the negative control group.A high value of PI was also shown in the BSTCR groups,especially in the high dose group where the influence of cell proliferation was stronger than that in the positive control group.The levels of E_2 and P in the BSTCR groups of all dosages were significantly higher than those in the negative control group,and did not show any significant difference compared with those in the positive control group.Levels of FSHR and IGF-1 mRNA expression in the BSTCR groups increased in a dose-dependent manner at levels higher than those in the negative control group.Conclusion:S-BSTCR can obviously stimulate the proliferation and steroidogenesis of ovarian GCs.It is speculated that BSTCR could play a regulatory action on ovarian function through two different pathways of endocrine and autocrine by promoting FSHR and IGF-1 mRNA expression.     

Effect of Atorvastatin on Serum MMP-2,MMP-9 and TIMP-1 in Rabbits with Chronic Heart Failure

Objective: To observe the effects of atorvastatin on serum matrix metalloproteinase-2(MMP-2), matrix metalloproteinase-9(MMP-9), and the tissue inhibitor of metalloproteinase-1(TIMP-1) in the development of chronic heart failure. To investigate the role of atorvastatin in the therapy of chronic heart failure and determine its possible mechanism of action. Methods: Thirty Japanese Big Ear rabbits were randomly selected and divided into 3 groups: sham-operated group(SO group), heart failure control group(HC group) and heart failure atorvastatin therapy group(HA group), with 6, 12 and 12 animals in the respective groups. Volume overloading was produced in the HC group and HA group animals by creating an aortic insufficiency, induced by damaging the aortic valve with a catheter introduced through the carotid artery. After 14 days, abdominal aorta constriction was performed in order to obtain a pressure overload. Six weeks later rabbits in the HA group were administered atorvastatin 3mg. Kg-1.d-1 for 4 weeks, at which time the experiment was terminated. Arterial blood was drawn and serum levels of MMP-2, MMP-9 and TIMP-1 were measured in all groups at the same time using an ELISA method. Results: Structural and functional indicators of chronic heart failure(CHF) were seen in both the HC and HA groups, but atorvastatin significantly reduced the observed effects. The serum concentrations of MMP-2, MMP-9 and TIMP-1 were at low levels in all three groups at the start of the study, with no difference between them(P < 0.05). At the end of 6th week concentrations were significantly increased in the HC and HA groups compared with the SO group(P < 0.05), but there were no differences between the HC group and HA group(P > 0.05). The increased concentrations in HC group continued to the end of the experiment, but values in the HA group were all lower than those in the HC group by the end of the experiment(P < 0.05). Conclusion: Serum concentrations of MMP-2, MMP-9 and TIMP-1 increase significantly during the course of CHF, paralleling the pathological progress of CHF. Atorvastatin benefits CHF, and the decreased serum levels of MMP-2, MMP- 9 and TIMP-1 may be one of the drug's mechanism of action.