Change of p16~(INK4a) and PNCA Protein Expression in Myocardium after Injection of hIGF-1 Gene Modified Skeletal Myoblasts into Post-infarction Rats

This study examined the change of p16^INK4a and PNCA protein expression in myocardium after injection of hIGF-1 gene modified skeletal myoblasts into post-infarction rats. HIGF-1 gene modified skeletal myoblasts （hIGF-1-myoblasts） were injected into hind limb muscles of 18 post-infraction rats （experimental group）. Primary-myoblasts were injected into 18 post-infraction rats （control group） and 12 non-infarction rats （sham group）. Expression of p16INK4a and PCNA pro- tein in myocardiums were separately detected immunocytochemically 1, 2 and 4 weeks after the inuection. The level of hIGF-1 and rIGF-1 protein in serum and myocardium were detected by en- zyme-linked immunosorbent assay （ELISA）. Compared with the sham group, the percentage of p^16INK4a and PCNA positive cells reached a peak after 1 week in the control group and the experimental group （P〈0.01）. Moreover, the percentage of p16^INK4a-positive cells in the experimental group was lower than in control group whereas the percentage of PCNA-positive cells was lower in the control group than in the experimental group （P〈0.01）. The percentage of p16^INK4a-positive cells in the experimental group and the percentage of PCNA-positive cells in the control group were close to that in the sham group from the 2nd week （P〉0.05）. ELISA analysis disclosed that the myocardium level of rIGF-1 protein increased gradually in the controls and especially in the experimental group （P〈0.01）. The serum level of rIGF-1 decreased significantly in post-infraction rats, but these conditions were improved in the experimental group （P〈0.01）. The hIGF-1 protein in serum and myocar- dium were detected from the 1st week to the 4th week in the experimental group. Statistical analysis revealed significant associations of myocardium level of hIGF-1 protein with expression of p^16INK4a and PCNA protein （r=–0.323, P〈0.05; r=0.647, P〈0.01）. It is concluded that genetically hIGF-1-myoblast provides a means for constant synthesis and re     

Myocardial autophagy variation during acute myocardial infarction in rats: the effects of carvedilol

Background The loss of cardiac myocytes is one of the mechanisms involved in acute myocardial infarction （AMI）-related heart failure. Autophagy is a common biological process in eukaryote cells. The relationship between cardiac myocyte loss and autophagy after AMI is still unclear. Carvedilol, a non-selective α1-and β-receptor blocker, also suppresses cardiac myocyte necrosis and apoptosis induced by ischemia. However, the association between the therapeutic effects of carvedilol and autophagy is still not well understood. The aim of the present study was to establish a rat model of AMI and observe changes in autophagy in different zones of the myocardium and the effects of carvedilol on autophagy in AMI rats. Methods The animals were randomly assigned to a sham group, an AMI group, a chloroquine intervention group and a carvedilol group. The AMI rat model was established by ligating the left anterior descending coronary artery. The hearts were harvested at 40 minutes, 2 hours, 24 hours and 2 weeks after ligation in the AMI group, at 40 minutes in the chloroquine intervention group and at 2 weeks in other groups. Presence of autophagic vacuoles （AV） in the myocytes was observed by electron microscopy. The expression of autophagy-, anti-apoptotic- and apoptotic-related proteins, MAPLC-3, Beclin-1, Bcl-xl and Bax, were detected by immunohistochemical staining and Western blotting. Results AVs were not observed in necrotic regions of the myocardium 40 minutes after ligation of the coronary artery. A large number of AVs were found in the region bordering the infarction. Compared with the infarction region and the normal region, the formation of AV was significantly increased in the region bordering the infarction （P 〈0.05）. The expression of autophagy- and anti-apoptotic-related proteins was significantly increased in the region bordering the infarction. Meanwhile, the expression of apoptotic-related proteins was significantly increased in the infarction region. In the chloroquine intervention group, a large number of initiated AVs （AVis） were found in the necrotic myocardial region. At 2 weeks after AMI, AVs were frequently observed in myocardial cells in the AMI group, the carvedilol group and the sham group, and the number of AVs was significantly increased in the carvedilol group compared with both the AMI group and the sham group （P 〈0.05）. The expression of autophagy- and anti-apoptotic-related proteins was significantly increased in the carvedilol group compared with that in the AMI group, and the positive expression located in the infarction region and the region bordering the infarction. Conclusions AMI induces the formation of AV in the myocardium. The expression of anti-apoptosis-related proteins increases in response to upregulation of autophagy. Carvedilol increases the formation of AVs and upregulates autophagy and anti-apoptosis of the cardiac myocytes after AMI.     

Huoxue Anxin Recipe(活血安心方)Promotes Myocardium Angiogenesis of Acute Myocardial Infarction Rats by Up-Regulating miR-210 and Vascular Endothelial Growth Factor

Objective: To investigate the microR NAs(miR NAs) expression profile of acute myocardial infarction(AMI) rats and the regulating effects of Huoxue Anxin Recipe(活血安心方, HAR) on angiogenesis-related miR NAs and genes. Methods: Forty-five Wistar rats were randomly assigned to 3 groups according to a random number table: sham, AMI, and AMI+HAR groups(15 in each group). AMI rats were established by ligation of the left descending coronary artery. HAR was intragastrically administered to rats of the AMI+HAR group for successive 21 days since modeling, meanwhile the same volume of 0.9% normal saline was administered to rats of the sham and AMI groups. Doppler echocardiography was used for noninvasive cardiac function test. Hematoxylin and eosin staining was used to observe the histopathological change. miR NAs expression profile was detected by quantitative realtime polymerase chain reaction(qR T-PCR). The mR NA and protein expressions of vascular endothelial growth factor(VEGF), and a target gene of miR-210 was further detected by qR T-PCR and Western blot, respectively. The microvessels density of myocardium was evaluated by CD31 immunostaining. Results: Compared with the sham group, ejection fraction(EF) and fractional shortening(FS) values were decreased significantly in the AMI group(P0.01), while the infarction area and the interstitial collagen deposition were increased obviously. As for the AMI+HAR group, EF and FS values were increased significantly(P0.05 vs. AMI group), and the infarction area was reduced and the interstitial collagen deposition were alleviated significantly. Total of 23 miR NAs in the AMI group expressed differently by at least 1.5 folds compared with those in the sham group; 5 miR NAs in the AMI+HAR group expressed differently by at least 1.5 folds compared with those in the AMI group. Among them, miR-210 was low in the AMI group and high in the AMI+HAR group. The relative mR NA and protein expressions of VEGF were decreased significantly in the AMI group(P0.05 vs. sham group), and increased significantly in the AMI+HAR group(P0.01 vs. AMI group). CD31 expression area and optical intensity were decreased significantly in the AMI group(P0.05 vs. sham group), and increased significantly in the AMI+HAR group(P0.01 vs. AMI group). Conclusions: HAR could reduce the infarction area, alleviate the interstitial fibrosis and improve the cardiac function of AMI rats. Those effects could be related to promoting myocardium angiogenesis of HAR by up-regulating miR-210 and VEGF.     

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.     

山茱萸提取物-总三萜烯酸改善链脲佐菌素诱导糖尿病大鼠肾损害进展

Objective:To Investigate whether total triterpene acids(TTAs),isolated from Corpus Fructus,attenuates renal function by reducing oxidative stress and down-regulating the expression of transforming growth factor β_1(TGF-β_1).Methods:Diabetes was induced by an injection of streptozotocin(40 mg/kg intravenously).Thirty rats were randomly divided into three groups:control group,diabetic model group and TTAs treatment group(50 mg/kg,intragastrically)administrated for 8 weeks from 5th to 12th week.All rats were anaesthetized and then were killed to remove kidneys.The renal function and redox enzyme system parameters were tested.Glomerular morphology was observed by a light microscopy.Immunohistochemistry and Western blot assays were employed to determine the protein levels of TGF-β_1.Results:TTAs attenuated the levels of urinary protein,serum creatinine and blood urea nitrogen,although it did not significantly reduce the level of glucose.In addition,TTAs decreased the malondialdehyde while increased superoxide dismutase,catalase and glutathione peroxide activities in diabetic rats.The renal pathological changes in TTAs treatment group were ameliorated.Furthermore,TTAs also ameliorated the expression of TGF-β_1.Conclusion:TTAs improved renal function via reducing oxidative stress and down-regulation the expression of TGF-β_1 in diabetic rats.     

High-frequency ultrasound evaluation of effects of early treatment with metoprolol on myocardial inflammatory cytokine expression in rats with acute myocardial infarction

This study evaluated the effects of early treatment with β-adrenergic blocker metoprolol on ventricular remodeling and function after acute myocardial infarction (AMI) by using high frequency ultrasound.The relationship between the efficacy and the expression level of cardiac myocardial inflammatory cytokine was examined in rats.The rat model of AMI was induced by ligating the left ante-rior descending artery.The surviving rats were randomly assigned to two experimental groups:MI control (MI) group and MI metoprolol (MI-B) group,with the rats undergoing sham operation serving as normal control (Sham).MI-B group was given metoprolol for 4 weeks (refer to the CCS-2 protocol) and the other two groups received equal volume of saline via intragastric (i.g.) administation.The ventricular remodeling and function were evaluated by high frequency ultrasound 4 weeks after the treatment.Then all rats were sacrificed for pathological examination and immunohistochemistrical detection of inflammatory cytokines,including IL-1β,IL-6,IL-10 and TNF-α.Compared with the MI group,the left ventricular end-systolic dimension,end-diastolic dimension,end-systolic volume and end-diastolic volume of the MI-B group were significantly decreased (P<0.01),while the left ventricular anterior wall end-diastolic thickness,ejection fraction and fractional shortening were obviously increased (P<0.01).The conspicuous improvement in the left ventricular morphology and function was coincident with the markedly reduced TNF-α and IL-1β expression and the increased IL-10 expression.We are led to conclude that early metoprolol treatment for AMI can regulate myocardial inflammatory cytokine expression to improve cardiac function and the underlying mechanism might be that it decreases the level of pro-inflammatory cytokines and increases the level of its anti-inflammatory counterparts in cardiac myocytes.Our study also showed that echocardiography is a useful technique for the structural and functional assessment of left ventricle after acute my     

Shenfu Injection(参附注射液) Suppresses Inflammation by Targeting Haptoglobin and Pentraxin 3 in Rats with Chronic Ischemic Heart Failure

Objective:To investigate the regulatory effects of Shenfu Injection(SFI,参附注射液) on hemodynamic parameters and serum proteins in rats with post-infarction chronic heart failure(CHF).Methods:Forty-five healthy Wistar rats were randomized into three groups:sham,heart failure(model) and SFI group.The CHF was induced by left coronary artery ligation.Seven days after the surgical operation,animals in the sham group and the model group received saline(6.2 mL/kg/d),while animals in the SFI group received SFI(6.2 mL/kg·d) intraperitoneally.Four weeks later,cardiac hemodynamic parameters were measured via the carotid route.The expression of serum proteins was analyzed by two-dimensional electrophoresis and matrixassisted laser desorption/ionization time-of-flight mass spectrometer(MALDI-TOF MS).Results:Recording of hemodynamic parameters showed that left ventricular systolic pressure(LVSP),maximum rate of left ventricular pressure(+dp/dt_(max)) rise,and maximum rate of left ventricular pressure(-dp/dt_(max)) decrease,while the left ventricular end diastolic pressure(LVEDP) rose in the model group compared to those in the sham group(P0.05).The results of the MALDI-TOF MS indicated that haptoglobin(HP),pentraxin 3(PTX3) and alpha-1-antitrypsin were up-regulated,while serum albumin and 40 S ribosomal protein were down-regulated in the model group(P0.05).Compared with the model group,LVSP,+dp/dt_(max)and-dp/dt_(max) were higher,while LVEDP was lower in the SFI group(P0.05).Expression levels of HP and PTX3 were lower than in the model group(P0.05).Conclusion:SFI could improve hemodynamic function and decrease inflammatory reactions in the pathophysiology of CHF.The serum proteins HP and PTX3 could be potential biomarkers for chronic ischemic heart failure,and they could also be the serum protein targets of SFI.     

Comparison of three doses of enalapril in preventing left ventricular remodeli ng after acute myocardial infarction in the rat

Objective To compare the effects of high, middle and low doses of enalapril in preventing left ventricular remodeling (LVRM) after acute myocardial infarction (AMI) in ra ts, especially evaluating the efficacy of low dose enalapril.Methods AMI was induced by ligating the left coronary artery in 149 female SD rats. 48 hours after the procedure, the 97 surviving rats were randomized to one of the f ollowing four groups: (1) AMI controls (n=24),(2) high-dose (10mg·kg(-1)·d(-1), n=25), (3) middle-dose (1mg·kg·d［-1］, n=23), and (4) low-dose (0.1mg·kg(-1)·d(-1), n=25) enalapril groups. I n addition, sham-operated (n=13) and normal rats (n=10) were randomly selected to serve as non-infarction controls. Enalapril was delivered by direct gastric gavage. After 4 weeks of therapy, hemodynamic studies were performed, then the rat hearts were fixed with 10% formalin and pathology analysis was performed. Exclusive of the dead rats and those with MI size &lt;35% or &gt;55%, complete experim ental data were obtained from 67 rats, which were comprised of (1) AMI controls (n=13), (2) high-dose enalapril (n=13), (3)middle-dose enalapril (n=12), (4) l ow-dose enalapril (n=12), (5) sham-operated (n=8) and (6) normal (n=9) groups .Results There were no significant differences among the four AMI groups in infarction si ze (all P&gt;0.05). Compared with the sham-operated group, the left vent ricu lar (LV) end diastolic pressure (LVEDP), volume (LVV), absolute and relative wei ght (LVAW, LVRW) in AMI group were all significantly increased (all P&lt;0.001 ), while maximum LV pressure rising and dropping rates (±dp/dt) and their corre cted values by LV systolic pressure (±dp/dt/LVSP) were all significantly reduce d in the AMI control group (P&lt;0.01-0.001), indicating LVRM occurred and LV systoli c and diastolic functions were impaired. Compared with the AMI group, LVEDP, LV V, LVAW and LVRW were all significantly decreased in the three enalapril groups (control P&lt;0.001), with the reduction of LVEDP, LVV and LVAW being mo re significant in high-dose than in low-dose enalapril groups (all P&lt;0.05 ), and the ±dp/dt/LVSP were significantly increased only in the high and middle -dose enalapril groups (P&lt;0.01).Conclusions High, middle and low doses of enalapril were all effective in preventing LVRM af ter AMI in the rat, with low dose enalapril being effective and high dose superi or. As for LV functional improvement, only high and middle-dose enalapril were effective.     

Effect of ETA receptor antagonist BQ-123 on cardiac function and endothelin system after coronary microembolization in rats

Objective To evaluate the effects of BQ-123 on cardiac function and ventricular remodelling after coronary microembolization (CME) in rats. Methods We created a rat model of CME by injecting a suspension of autogenic microthrombotic particles into left ventricle. Three days after the procedure, the 30 surviving rats were randomly divided into 3 groups, each consisted of 10 rats: sham-operation group(SO), CME model group(CM) and BQ-123 intervention group(BQ). Rats in the BQ group received BQ-123 (400μg/kg per day, intraperitoneally) for 4 weeks. Plasma and myocardial endothelin-1 (ET-1) were measured by radioimmunoassay. And serial echocardiography was performed to monitor alterations of left ventricular end-systolic and end-diastolic diameter (LVESD, LVEED), and left ventricular short-axis fraction shortening(LVFS) and ejection fraction (LVEF), and physiologicography to document the changes of left ventricular systolic pressure (LVSP) and end-diastolic pressure pressure(LVEDP), and left ventricular maximum positive and negative dp/dt (±LVdp/dtmax). Results Compared with sham-operated group, both LVEDD and LVESD were increased (P<0.01), whereas LVFS and LVEF were significantly decreased (P<0.01) in CME group; LVEDP was markedly increased, while LVSP and±LVdp/ dtmax markedly reduced in CME group (P<0.01); plasma and tissue ET-1 levels increased in CME group (P<0.01). BQ-123 intervention significantly decreased both the plasma and tissue ET-1 levels (P<0.01), and markedly increased LVFS and LVEF, with significant improvement of LVSP and±LVdp/ dtmax (P<0.01). Conclusions Treatment with BQ-123 prevents ventricular remodeling after CME due to suppression of the endothelin system.     

Effects of the Drug(BSZGC)--containing Serum on Proliferation of Rat''s Osteoclasts and TRACP Activity in vitro

Objective:To observe effects of the drug-containing serum of Bu Shen Zhuang Gu Capsule(BSZGC 补肾壮骨胶囊 Capsule for Tonilying the Kidney to Strengthen the Bones)on proliferation of the rat's osteoclasts and tartrate-resistant acid phosphatase (TRACP)activity in vitro SO as to delve into the mechanisms of its preventive and therapeutic actions on osteoporosis.Methods:Forty female Sprague.Dawley rats aged three months were randomly divided into high dosage BSZGC group,medium dosage BSZGC group,low dosage BSZGC group,and the control group.BSZGC was orally administered into the rats of high,medium,and low dosage groups at difierent dosages for 12 days.and isometric normal saline was orally administered to the rats of the Control group.The drug-containing serum and control serum were prepared.Osteoclasts isolated mechanically from the femur and tibia of Sprague-Dawley rats aged one week were cultured wim medium added with different drug-containing sera and control serum.The growth of osteoclasts was observed under an inverted phase-contrast microscope,and optic density(OD)value of osteoclasts was determined by MTT colorimetric assay.TRACP activity was measured by the diazol method.Results:OD value of osteoclasts in the high dosage drug-containing serum group,medium dosage drug-containing serum group,and low dosage drug-containing serum group was significantly lower than that in the control serum group(P＜0.05)with a dose-effect correlation.TRACP activity in high dosage drug-containing serum group,medium dosage drug-containing serum group,low dosage drug-containing serum group was significantly lower than that of the control serum group(P＜0.01),and no significant differences in TRACP activity were not found among the difierent dosages drug-containing serum groups.Conclusions:BSZGC can inhibit the proliferation of the osteoclasts and reduce TRACP activity,which may be one of the mechanisms of its preventive and therapeutic actions on osteoporosis.     

Huannao Yicong Decoction(还脑益聪方) Extract Reduces Inflammation and Cell Apoptosis in Aβ_(1-42)-Induced Alzheimer's Disease Model of Rats

Objective: Huannao Yicong Decoction(还脑益聪方, HYD), an effective herbal formula against Alzheimer's disease(AD), has been proven to have neuroprotective action in amyloid β-protein_(1-42))(Aβ_(1-42))-induced rat model. This study was designed to characterize mechanisms by which HYD leads to suppression of inflammation and apoptosis in the brains of Aβ_(1-42)-induced rat. Methods: A total of 72 rats were divided into 6 groups, which were referred to as: sham operation group, model group, donepezil-treated group, HYD low-dose group(HYDL), HYD middle-dose group(HYDM) and HYD high-dose group(HYDH). Rats in HYDL, HYDM and HYDH were injected with Aβ_(1-42) at the CA1 region of hippocampus to form AD model and were fed the HYD extract at different dose of 3.78, 7.56 and 18.90 g crude drug/kg. The behavioral changes of rats were evaluated by Morris water maze(MWM) before sacrifice. Pathological changes of the brain tissue were evaluated using hematoxylin eosin(HE) staining. The levels of interleukin-1(IL-1) and tumor necrosis factor-α(TNF-α) were measured by radioimmunoassay. The levels of Aβ and proteins that are associated with apoptosis such as B-cell lymphoma-2 protein(Bcl-2), Bcl-2-associated X protein(Bax), cysteine-aspartic protease(caspase)-3,-8,-9 and-12 in serum were measured by immunohistochemistry. Results: Compared with the sham operation group, the spatial learning and memory abilities of AD rats were significantly decreased(P0.05 or P0.01; Expressions of IL-1, TNF-α, Aβ and apoptosis-signaling proteins caspase-3,-8,-9,-12 were significantly up-regulated(P0.05 or P0.01). The ratio of Bcl-2 to Bax were significantly decreased in the model group(P0.01). When treated with HYD extract, the spatial learning and memory abilities of AD-model rats were significantly increased(P0.05 or P0.01), IL-1, TNF-α, Aβ, caspase-3,-8,-9 and-12 were down-regulated(P0.05 or P0.01), and the ratio of Bcl-2 to Bax were reduced(P0.05 or P0.01). Conclusions: HYD extract can improve the learning and memory ability deficits, alleviate the inflammatory response and pathological manifestations induced by Aβ_(1-42) injection in the rat model of AD. HYD down-regulates the levels of IL-1, TNF-α and Aβ, and decreases the rate of apoptosis by modulating apoptosis-signaling-related proteins such as caspase-3,-8,-9, and-12.     

Effects of Bone Marrow-derived Mesenchymal Stem Cells on TGF-β and MCP-1 in Injured Lung of Rats

The primary objective of the present study is to investigate the therapeutic effect of bone marrow-derived mesenchymal stem cells （MSCs） on bleomycin （BLM）-induced lung injury of rats and the effect on transforming growth factor-β （TGF-β） and monocyte chemoattractant protein-1 （MCP-1）. MSCs were isolated from SD rats. The recipients rats were divided randomly into four groups： lung injury group, MSC treatment group, MSC control group and normal control group. Rats of lung injury group and MSC treatment group were perfused with BLM of 5mg/kg （0.2-0.3ml） intratracheally, others were perfused with normal saline. After twelve hours, rats of MSC treatment group and MSC control group were injected MSCs of 0.5×10^6per rat into tail vein. Haematoxylin-eosin staining was used to observe the morphology in lung tissue. ELISA was used to detect the contents of TGF-β and MCP-1 in serum and bronchoalveolar lavage fluid （BALF）. Collagen content of the lung tissue was assessed by hydroxyproline （HYP） concentration. It was found that the thickness of alveolar wall and lung interstitium were significantly reduced in the rats of MSC treatment group compared with the lung injury group. HYP content in lung interstitium, TGF-β and MCP-1 in serum and BALF were increased significantly in rats of lung injury group two weeks after BLM perfusion, but they were reduced significantly in the rats of MSC treatment group compared with the injured rats. These observations provide evidence that MSCs engraftment could alleviate bleomycin-induced lung injury and fibrosis in rats and the therapeutic effects might relate with the decrease of TGF-β and MCP-1.     

Effect of TGF-β1 on the Expression of IL-12, IL-15, IL-18, IL-4 and IL-10 in Heart Transplantation Rejection in Rats

To investigate the effect of TGF-β1 on the expressions of IL-12, IL-15, IL-18, IL-4 and IL-10 in heart transplantation rejection in rats, a model of rat cervical heterotopic heart transplantation was set up and the model rats were randomly divided into three groups： control group, transplant group and TGF-β1 group. The mRNA expression levels of IL-12, IL-15, IL-18, IL-4 and IL-10 were determined by RT-PCR at the 5th day after the transplantation. The mRNA expression levels of IL- 12, IL-15, IL-18 were increased obviously and those of IL-4, IL-10 were significantly decreased in the transplant group as compared with the control group （P〈0.01）. In the TGF-β1 group, the mRNA ex- pression levels of IL- 12, IL- 15, IL- 18 were significantly decreased and those of IL-4, IL- 10 were significantly increased as compared with the transplant group （P〈0.01）. The immunosuppressive effect of TGF-β1 on heart transplantation rejection was related to its inhibition of the expressions of Th1-type cytokines （IL-12, IL-15, IL-18 etc） and its promotion of the expressions of Th2-tpye cyto- kines （IL-4, IL-10）.     

Effect and mechanism of panaxoside Rg1 on neovascularization in myocardial infarction rats

<正>Objective:To investigate the effects and mechanisms of panaxoside Rg1 on the new vessel formation in acute myocardial infarction(AMI) rats.Methods:The AMI model of male Sprague-Dawley(SD) rats was established,and rats were randomly divided into the AMI model group,the treatment group of panaxoside Rg1,the placebo group and the treatment group of panaxoside Rg1 plus rapamycin.Cardiac creatases were determined with 1 mL blood drawn from vena caudalis of the rats 48 h after the model was successfully made. After 4 weeks,Evans blue was injected into the aorta roots of the rats,and then,red tetrazoline was dyed again and the myocardial infarction area was evaluated.The microvessel density(MVD) of infarction area was determined by the immunohistochemistry of CD31;enzyme-linked immunosorbent assay(ELISA) was used to detect the protein content of CD31 and hypoxia inducible factor-1α(HIF-1α) of the infarction area. Results:The MVD in the infarction area and the contents of CD31 and HIF-1αin the Rg1 treatment group were higher than those in the AMI model group significantly(P0.05).The cardiac creatase and infarction area were lower in the Rg1 treatment group than those in the AMI model group significantly(P0.05).The above effects,however,disappeared when rapamycin,the antagonist of mammalian target of rapamycin(mTOR),was administered simultaneously.Conclusions:Panaxoside Rg1 could increase the expression of HIF-1αand CD31 of myocardium and stimulate the angiogenesis.The above mentioned role of panaxoside Rg1 might be related to the excitation of mTOR receptor.     

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.     

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.     

Protective Effect of Norcantharidin on Collagen-Induced Arthritis Rats

Objective: To observe the effect of norcantharidin(NCTD) on collagen-induced arthritis(CIA) rats. Methods: Sixty Sprague-Dawley(SD) rats were randomly divided into 6 groups(n=10): normal group, CIA model group(model group), NCTD low-dose group [1.35 mg/(kg·d)], NCTD middle-dose group [2.7 mg/(kg·d)], NCTD high-dose group [5.4 mg/(kg·d)] and methotrexate(MTX) group [1.8 mg/(kg/w)]. Anesthetized rats were sacrificed by luxation of cervical vertebra after 4 weeks of administration. The arthritis scores were evaluated twice a week. The pathological changes in the ankle joints of rats were observed by hematoxylin-eosin(HE) staining. The serum levels of interleukin(IL) 1β, IL-6, tumor necrosis factor(TNF)-α, vascular endothelial growth factor(VEGF), IL-17 and transform growth factor(TGF) β were detected by enzyme linked immunosorbent assay(ELISA). The mRNA expression of retinoid-related orphan nuclear receptor γ t(ROR γ t) and forkhead box P3(Foxp3) in peripheral blood lymphocytes were confirmed by real-time polymerase chain reaction. Results: MTX and high-dose NCTD not only decreased the arthritis scores but also alleviated the pathological changes in CIA rats' ankle joints compared with the model group(P0.05 or P0.01). All doses of NCTD significantly inhibited the serum levels of IL-6, IL-17 and TNF-α in CIA rats(P0.05). Only middle-and high-dose of NCTD prominently decreased serum IL-1β and TGF-β levels of CIA rats(P0.05). However, NCTD has no effect on vascular endothelial growth factor(VEGF) level in CIA rats. The Foxp3 mRNA expression in all NCTD groups were increased significantly than in the model group(P0.05). The mRNA expression of RORγt in NCTD high-dose group was decreased apparently in comparison with the model group(P0.05). Conclusion: NCTD showed therapeutic effect on CIA rats by inhibition of cytokines and regulation of Th17/Treg cells.     

Expression of Serum and Glucocorticoid-inducible Kinase1 in Diabetic Rats and Its Modulation by Fluvastatin

The expression of serum and glucocorticoid-induced protein kinase in the renal cortex of diabetic rats was examined, and the function of signal transduction mediated by SGK1 in diabetic nephropathy and its modulation by fluvastatin were also investigated. 24 male Wistar rats were randomly divided into normal control group （n = 8）, diabetic nephropathy group （n = 8） and fluvastatin-treated diabetic nephropathy group （15 mg/kg/d, n=8）. The metabolic parameters were measured at the 8th week. The expression of transforming growth factor β1 （TGF-β1） and fibronectin （FN） was immunohistochemically examined. The expression of SGK1 was detected by RT-PCR and Western blot, and CTGF mRNA was assessed by RT-PCR. As compared to DN, blood glucose, 24-h urinary protein, Cer and kidney weight index were all decreased and the weight was increased obviously in group F. At the same time, mesangial cells and extracellular matrix proliferation were relieved significantly. The levels of cortex SGK1 mRNA and protein were up-regulated, and both TGF-β1 and FN were down-regulated by fluvastatin. The mRNA of SGK1 was positively correlated with the CTGF, TGF-β1 and FN. SGK1 expression is markedly up-regulated in the renal cortex of DN group and plays an important role in the development and progress of diabetic nephropathy by means of signal transduction. Fluvastatin suppressed the increased SGKlmRNA expression in renal cortex and postponed the development of diabetic nephropathy.