Effects of clenbuterol exposure on PPARγ expression in adipocytes of rats

Objective To investigate effects of clenbuterol(CLB) on the peroxisome proliferatorsactivated receptor γ(PPARγ) expression in adipose tissues of rats.Methods Twenty adult female Sprague-Dawley rats were randomly divided into4 groups(5 rats per group). CLB solved in normal saline solution was given at the dose of 0 mg/kg body weight(bw)(group A, as the control), 0.4 mg/kg bw(group B,low-dose group), 2.0 mg/kg bw(group C, mid-dose group), and 18.5 mg/kg bw(group D,high-dose group) for 14 d by gavage consecutively, respectively. Methods of immunohistochemistry, quantitative Real-time PCR and Western blotting were performed to detect expression of PPARγ in the adipose tissue samples.Results PPARγ-positive immunostaining was strong in the controls and weak in the experimental groups. There was no difference on PPARγ mR NA and protein between the low-dose group and the control(P>0.05). With the increase of CLB doses, expression levels of PPARγ mR NA and protein were significantly lower in mid- or high-dose group than those in the control(P<0.01).Conclusions The PPARγ expression in adipose tissues of rats could be down-regulated after CLB exposure, and the decrease became more severe with the increasing doses.     

Mitofusin2 decreases intracellular cholesterol of oxidized LDL-induced foam cells from rat vascular smooth muscle cells

Mitofusin2 (Mfn2) plays a pivotal role in the proliferation and apoptosis of vascular smooth muscle cells (VSMCs). The purpose of this study was to investigate the effects of Mfn2 on the trafficking of intracellular cholesterol in the foam cells derived from rat VSMCs (rVSMCs) and also to investigate the effects of Mfn2 on the expression of adenosine triphosphate-binding cassette subfamily A member 1 (ABCA1), adenosine triphosphate-binding cassette subfamily G member 1 (ABCG1) and peroxisome proliferator-activated receptor gamma (PPARγ). The rVSMCs were co-cultured with oxidized low density lipoprotein (LDL, 80 μg/mL) to produce foam cells and cholesterol accumulation in cells. Before oxidized LDL treatment, different titers (20, 40 and 60 pfu/cell) of recombinant adenovirus containing Mfn2 gene (Adv-Mfn2) were added into the culture medium for 24 h to transfect the Mfn2 gene into the rVSMCs. Then the cells were harvested for analyses. The protein expression of Mfn2 was significantly higher in Adv-Mfn2-transfected group than in untransfected group (P<0.05), and the expression levels significantly increased when the titer of Adv-Mfn2 increased (P<0.05). At 24 or 48 h after oxidized LDL treatment, rVSMCs became irregular and their nuclei became larger, and their plasma abounded with red lipid droplets. However, the number of red lipid droplets was significantly decreased in Adv-Mfn2-transfected group as compared with untransfected group. At 48 h after oxidized LDL treatment, the intracellular cholesterol in rVSMCs was significantly increased (P<0.05), but it was significantly decreased in Adv-Mfn2-transfected group as compared with untransfected group (P<0.05), and it also significantly decreased when the titer of Adv-Mfn2 increased (P<0.05). The mRNA and protein expression levels of ABCA1 and ABCG1 were significantly increased in Adv-Mfn2-transfected group as compared with untransfected group (P<0.05). Though the mRNA and protein expression levels of PPARγ was not significantly increased (P>0.05), the phosporylation levels of PPARγ were significantly decreased in Adv-Mfn2-transfected group as compared with untransfected group (P<0.05). These results suggest that the transfection of Adv-Mfn2 can significantly reduce intracellular cholesterol in oxidized LDL-induced rVSMCs possibly by decreasing PPARγ phosporylation and then increasing protein expression levels of ABCA1 and ABCG1, which may be helpful to suppress the formation of foam cells.     

Effect of Schisandra Chinensis on Interleukins,Glucose Metabolism,and Pituitary-Adrenal and Gonadal Axis in Rats under Strenuous Swimming Exercise

Objective:To investigate the effect of Chinese medicine(CM) Schisandra chinensis on interleukin(IL),glucose metabolism,and pituitary-adrenal and gonadal axis of rats after strenuous navigation and exercise.Methods:A total of 45 Sprague-Dawley rats were randomized into the quiet control group,the stress group,and the CM group(15 in each group).The CM group received 2.5 g/kg of Schisandra chinensis twice per day for one week before modeling.Except the quiet controls,rats were trained using the Bedford mode for 10 days.On the11 th day,they performed 3 h of stressful experimental navigation and 3 h of strenuous treadmill exercise.The levels of serum testosterone(T),Cortisol(CORT),luteinizing hormone(LH),IL-1,IL-2,and IL-6 were tested by radioimmunoassay and enzyme-linked immunosorbent assay,respectively.The adrenal cortex ultrastructure was observed using electron microscopy.Results:Compared with the quiet control group,after navigation and strenuous exercise,blood glucose was increased,and T level was decreased in the stress group(both P<0.01).The blood glucose,CORT,IL-1 and IL-2 levels were significantly reduced in the CM group(P<0.05 or P<0.01) as compared with the stress group.Electron microscopy revealed that the rats in the CM group had a smaller decrease in adrenal intracellular lipid droplets and higher levels of apoptosis than those in the stress group.Conclusions:Schisandra chinensis can reduce serum CORT and blood glucose levels in stressed rats.It appears to protect the cell structure of the adrenal cortex,and offset the negative effects of psychological stress and strenuous exercise related to immune dysfunction.Schisandra chinensis plays a regulatory role in immune function,and can decrease the influence of stress in rats.     

Antidepressant-Like Effects of Chaihu Shugan Powder (柴胡疏肝散) on Rats Exposed to Chronic Unpredictable Mild Stress through Inhibition of Endoplasmic Reticulum Stress-Induced Apoptosis

Objective: To investigate the antidepressant-like effects of Chaihu Shugan Powder (CSP, 柴胡疏肝散) and to explore its underlying mechanisms. Methods: Thirty-two Sprague-Dawley rats were randomly divided into control (CON), chronic unpredictable mild stress (CUMS), fluoxetine (FLU), and CSP groups, 8 rats in each group. All of the rats except for those in the control group were subjected to 3 consecutive weeks of CUMS to establish the depression model. The open field test (OFT), forced swimming test (FST), and sucrose preference test were used to assess the anti-anxiety and antidepressant effects of CSP. Terminal deoxynucleotidyl transferase (TdT) dUTP nick-end labeling was used to determine the apoptosis rate in the hippocampal tissues. The mRNA and protein levels of glucose-regulated protein (GRP) 78, spliced X-box-binding protein (XBP)-1, CCAAT/enhancer binding protein homologous protein (CHOP), caspase-12, and c-Jun N-terminal kinase (JNK) in the hippocampus of rats were evaluated by real-time PCR and Western blot analysis, respectively. Results: Administration of CSP alleviated anxiety and depression-like behavior in CUMS rats, as revealed by enhanced time and distance in the center of the OFT (P<0.05), an increased preference for sucrose, and longer swimming time and shorter immobility time during the FST (all P<0.05). In addition, CSP treatment significantly reduced the rate of apoptosis in rat hippocampal neurons (P<0.05). The mRNA and protein expression levels of GRP78, spliced XBP-1, and CHOP were down-regulated along with the expression of caspase-12 and cleaved caspase-12 proteins (all P<0.05), whereas total and phosphorylated JNK1 protein levels did not differ significantly between control and CSP-treated rats. Conclusion: CSP can improve depression-like behavior in rats exposed to CUMS, possibly by suppressing CHOP and caspase-12 mediated apoptosis in the rat hippocampus.     

Pioglitazone ameliorates experimental nonalcoholic steatohepatitis by regulating hepatic nuclear factor-kappa B and cyclooxygenases-2 expression in rats

Background Pioglitazone is effective in nonalcoholic steatohepatitis (NASH), but the mechanism of action is not completely understood. This study was designed to investigate the impact of pioglitazone on hepatic nuclear factor-kappa B (NF-κB) and cyclooxygenases-2 (COX-2) expressions in NASH rats. Methods Thirty Sprague-Dawley male rats were randomly assigned to a control group (n=10), NASH group (n=10) and pioglitazone treatment group (n=10). Liver tissues were processed for histology by HE and Masson stained. Biochemical parameters of antioxidant enzyme activities, tumor necrosis factor alpha (TNF-α), prostaglandin E2 (PGE2) in the serum and hepatic samples were measured. The mRNA and protein expressions of peroxisome proliferator-activated receptor gamma (PPARγ), NF-κB and COX-2 were determined by real-time polymerase chain reaction, Western blotting and immunohistochemistry, respectively. Results There were severe steatosis, moderate inflammatory cellular infiltration and fibrosis in the livers of the NASH models. After treatment, steatosis, inflammation and fibrosis were significantly improved compared with the NASH group (χ2＝20.40, P＜0.001; χ2＝20.17, P＜0.001; χ2＝13.98, P=0.002). The serum and hepatic levels of total anti-oxidation competence (T-AOC), superoxide dismutase (SOD), catalase(CAT), glutathione peroxidase (GSH-PX) and malondialdehyde (MDA) in the NASH group were conspicuous disordered than those indicators in the control group. In the NASH group, the levels of serum TNF-α and PGE2 were significantly increased compared with the control group. Immunohistochemistry showed expressions of NF-κB and COX-2 in livers were significantly elevated, but PPARγ was decreased in the NASH group. Real-time PCR and Western blotting revealed mRNA and protein expressions of COX-2 were increased in the NASH group compared with the control group (0.57±0.08 vs 2.83±0.24; 0.38±0.03 vs 1.00±0.03, P＜0.001 and P=0.004, respectively). After pioglitazone intervention, all of those indicators markedly improved(P＜0.05 or P＜0.01 ). Conclusion Suppressing hepatic NF-κB and COX-2 expression, at least in part, is one of the possible therapeutic mechanisms of pioglitazone in NASH rats.     

EFFECT OF PEROXISOME PROLIFERATOR-ACTIVATED RECEPTOR ACTIVATORS ON TUMOR NECROSIS FACTOR-αL EXPRESSION IN NEONATAL RAT CARDIAC MYOCYTES A

Objecfive To investigate the effect ofperoxisome proliferator-αctivated receptor-α (PPARα) and PPARγ activators on tumor necrosis factor-α (TNFα) expression in neonatal rat cardiac myocytes. Primary cultures of cardiac myocytes from 1- to 3-dayold Wistar rats were prepared, and myocytes were ex-posed to lipopolysaccharide (LPS) and varying concentrations of PPARα or PPARγ activator (fenofibrate or pioglitazone).RT-PCR and ELISA were used to measure TNFα, PPARα, and PPARγ expression in cultured cardiac myocytes. Transient transfection of TNFα promoter with or without nuclear factor-kappaB (NF-κB) binding site to cardiac myocytes was performed. Performed Pretreatment of cardiac myocytes with fenofibrate or pioglitazone inhibited LPS-induced TNFα mRNA and protein expression in a dose-dependent manner. However, no significant changes were observed on PPARα or PPARα mRNA expression when cardiac myocytes were pretreated with fenofibrate or pioglitazone. Proportional suppression of TNFα promoter activity was observed when myocytes was transiently transfected with whole length of TNFα promoter (-721/ 17) after being stimulated with LPS and fenofibrate or pioglitazone, whereas no change of promoter activity was observed with transfection of TNFα reporter construct in deletion of NF-κBbinding site (- 182/ 17). Conchusions PPARα and PPARγ activators may inhibit cardiac TNFα expression but not accompanied by change of PPARα or PPARγ mRNA expression. Therefore PPARα and PPARγ activators appear to play a role in anti-inflammation.The mechanism may partly be involved in suppression of the NF-κBpathway.     

Effect of oxidative stress on ventricular arrhythmia in rabbits with adriamycin-induced cardiomyopathy

The purpose of the present study was to examine the effects of oxidative stress on ventricular arrhythmias in rabbits with adriamycin-induced cardiomyopathy and the relationship between oxidative stress and ventricular arrhythmia. Forty Japanese white rabbits were randomly divided into four groups (n=10 in each): control group, metoprolol (a selective β1 receptor blocker) group, carvedilol (a nonselective β blocker/α-1 blocker) group and adriamycin group. Models of adriamycin-induced car-diomyopathy were established by intravenously injecting adriamycin hydrochloride (1 mg/kg) to rabbits via the auri-edge vein twice a week for 8 weeks in the adriamycin, metoprolol and carvedilol groups. Rabbits in the control group were given equal volume of saline through the auri-edge vein. Rabbits in the metoprolol and carvedilol groups were then intragastrically administrated metoprolol (5 mg/kg/d) and carvedilol (5 mg/kg/d) respectively for 2 months, while those in the adriamycin and control groups were treated with equal volume of saline in the same manner as in the metroprolol and carvedilol groups. Left ventricular end diastolic diameter (LVEDd) and left ventricular ejection fraction (LVEF) were measured by echocardiography. Plasma levels of N-terminal pro B-type natriuretic peptide (NT-proBNP), malondialdehyde (MAD) and superoxide dismutase (SOD) were detected. The left ventricular wedge preparations were perfused with Tyrode’s solution. The transmural electrocardiogram, transmural action potentials from epicardium (Epi) and endocardium (Endo), transmural repolarization dispersion (TDR) were recorded, and the incidences of triggered activity and ventricular arrhythmias were obtained at rapid cycle lengths. The results showed that TDR and the serum MDA and NT-proBNP levels were increased, and LVEF and the serum SOD level decreased in the adriamycin group compared with the control group. The incidences of triggered activity and ventricular arrhythmia were significantly higher in the adriamycin group than those in the control group (P<0.05). In the carvedilol group as compared with the adriamycin group, the serum SOD level and the LVEF were substantially increased; the TDR, and the serum MDA and NT-proBNP levels were significantly decreased; the incidences of triggered activity and ventricular arrhythmia were obviously reduced (P<0.05). There were no significant differences in the levels of MDA and SOD, LVEF, TDR and the incidences of triggered activity and ventricular arrhythmia between the adriamycin group and the metoprolol group. It was concluded that carvedilol may inhibit triggered activity and ventricular arrhythmias in rabbit with adriamycin-induced cardiomyopathy, which is related to the decrease in oxygen free radials.     

Role of ghrelin in cognitive impairment of hypertensive elders with chronic psychological distress

Objective： To investigate ghrelin level change in combination with psychological stress in the hypertensive old people with cognitive impairment and to explore its effect as well as possible mechanism. Methods： The study population of 300 elders was divided into 2 groups, 148 with hypertension and 152 non-hypertension, who were screened for psychological distress and cognition function, and had blood drawn to measure plasma levels of ghrelin and total cortisol on the same day. Results： The rates of anxiety and cognitive impairment were higher in the hypertension elders, which were negatively correlated with plasma ghrelin level, resulted from chronic cortisol response to anxiety. Conclusion： Chronic plasma cortisol increase to long-term anxiety leads a reduce in ghrelin level which then adversely affects blood pressure and cognitive function in old people. So measuring ghrelin of elders may be a diagnostic tool to predict cognitive development and ghrelin may be a selective antihypertensive medicine for cognitive impairment elders with or without chronic psychological stress     

EFFECT OF PEROXISOME PROLIFERATOR-ACTIVATED RECEPTOR ACTIVATORS ON TUMOR NECROSIS FACTOR-α EXPRESSION IN NEONATAL RAT CARDIAC MYOCYTES

Objective To investigate the effect ofperoxisome proliferator-activated receptor-α (PPARα) and PPARγ activators on tumor necrosis factor-α (TNFα) expression in neonatal rat cardiac myocytes.Methods Primary cultures of cardiac myocytes from 1- to 3-day-old Wistar rats were prepared, and myocytes were exposed to lipopolysaccharide (LPS) and varying concentrations of PPARα or PPARγ activator (fenofibrate or pioglitazone).RT-PCR and ELISA were used to measure TNFα, PPARα, and PPARγexpression in cultured cardiac myocytes. Transient transfection of TNFα promoter with or without nuclear factor-kappaB (NF-κB) binding site to cardiac myocytes was performed.Results Pretreatment of cardiac myocytes with fenofibrate or pioglitazone inhibited LPS-induced TNFα mRNA and protein expression in a dose-dependent manner. However, no significant changes were observed on PPARα or PPARγ mRNA expression when cardiac myocytes were pretreated with fenofibrate or pioglitazone. Proportional suppression of TNFαpromoter activity was observed when myocytes was transiently transfected with whole length of TNFα promoter (-721/ 17) after being stimulated with LPS and fenofibrate or pioglitazone, whereas no change of promoter activity was observed with transfection of TNFα reporter construct in deletion of NF-κB binding site (-182/ 17).Conclusions PPARα and PPARγ activators may inhibit cardiac TNFα expression but not accompanied by change of PPARα or PPARγmRNA expression. Therefore PPARα and PPARγ activators appear to play a role in anti-inflammation.The mechanism may partly be involved in suppression of the NF-κB pathway.     

Tenascin-x facilitates myocardial fibrosis and cardiac remodeling through transforming growth factor-β1 and peroxisome proliferator- activated receptor γ in alcoholic cardiomyopathy

Background  Tenascin-x, an extracellular matrix glycoprotein exclusively expressed in fibroblasts, can mediate fibrosis in the presence of collagen. Therefore, we have investigated its potential role in facilitating myocardial fibrosis and cardiac remodeling via the transforming growth factor-β1 and peroxisome proliferator-activated receptor γ (TGFβ₁-PPARγ) pathway in alcoholic cardiomyopathy (ACM).

Methods  Experimental animals were divided into control (group A) and tenascin-x knock-out groups (group B) receiving alcohol. Six months post treatment, cardiac ejections fraction (EF), fractional shortening (FS), left ventricle end-diastole internal diameter (LVEDd) and collagen column fraction (CVF) were observed. Tenascin-x, smad-3, TGFβ₁, smad-7 and PPARγ protein expression levels were detected by Western blotting.

Results  Six months post treatment, EF and FS values were higher in group B than in group A (P <0.05 and P <0.01, respectively), while LVEDd and CVF were lower in group B (P <0.05 and P <0.01, respectively). Tenascin-x, smad-3 and TGFβ₁ protein expression levels were higher in group A, while smad-7 and PPARγ levels were lower than in group B (P <0.01), as measured by immunohistochemistry and Western blotting. Tenascin-x protein expression was negatively correlated with EF, FS, smad-7 and PPARγ, and positively correlated with LVEDd, CVF, smad-3, and TGFβ₁ (P <0.001).

Conclusion  Tenascin-x is an initiator of myocardial fibrosis and ACM development via upregulation of TGFβ₁ and downregulation of PPARγ.

     

Protective Effects of Hu-Lu-Ba-Wan (葫芦巴丸) against Oxidative Stress in Testis of Diabetic Rats through PKCα /NAPDH Oxidase Signaling Pathway

Objective: To explore the protective effect and the underlying mechanism of Hu-Lu-Ba-Wan (葫芦巴丸, HLBW) on the testis of diabetic rats. Methods: Twenty-four male Wistar rats (160–180 g) were randomly divided into 3 groups according to a random number table, including a control group (n=8), diabetic group (n=8), and HLBW group (n=8). Diabetic rat model was established by high-fat-diet administration and single intravenous injection of streptozotocin (26 mg/kg). Then HLBW granule was administrated for 12 weeks. Fasting blood glucose and insulin levels as well as serum total testosterone level and testicular testosterone content were examined. Oxidative stress markers in both serum and testis were tested. Meanwhile, testicular morphology was observed under hematoxylin and eosin (HE) and the ultrastructure of Leydig cell was observed by electron microscope. The superoxide anion level was detected by DHE, and TUNEL-positive cells of testis was evaluated by TUNEL assay. The gene and protein expression of protein kinase C (PKCα ), phosphorylated PKCα (P-PKCα ) and P47phox in testicular tissues were determined by quantitative RT-PCR analysis and Western bolt analysis. Results: Compared with the diabetic group, HLBW treatment significantly reduced the fasting glucose levels and increased the levels of fasting insulin and testosterone in serum (P<0.01). HLBW administration also reduced the levels of reactive oxygen species (ROS) in plasma and alleviated the damage of oxidative stress in the testis of diabetic rats. Additionally, HLBW down-regulated the protein and mRNA levels of PKCα , P-PKCα and P47phox in testicular tissues. Conclusion: HLBW may attenuate the oxidative stress in the testis of diabetic rats via PKCα /NAPDH oxidase signaling pathway.     

Xuefu Zhuyu Oral Liquid (血府逐瘀口服液) Prevents Apoptosis of Ischemic Myocardium Cells in Rats by Regulating SIRT1 and Its Pathway-Related Genes

Objective: To observe the changes of ischemic myocardial cells apoptosis in rats following intervention with Xuefu Zhuyu Oral Liquid(血府逐瘀口服液, XFZY), as well as changes of protein expression of silent information regulator 1(SIRT1) and SIRT1 pathway-related genes. Methods: H9c2 rat myocardial cells were divided into 6 groups: control group, oxygen glucose deprivation(OGD) group, SIRT1 siRNA group, OGD+SIRT1 siRNA group, OGD+XFZY group, and OGD+SIRT1 siRNA+XFZY group. Quantitative fluorescent polymerase chain reaction(PCR) and Western blot were used to detect the concentration variations of SIRT1 and its pathway-related genes and corresponding protein expression after XFZY intervention and SIRT1 transfection. Results: Compared with the control group, the m RNA and protein expressions of SIRT1 were decreased obviously, while the mRNA and protein levels of P53, forkhead box protein O1(FoxO1), FoxO3, FoxO4 and nuclear factor kappa B(NF-κB) were increased in the OGD group, SIRT1 siRNA group, and OGD+SIRT1 siRNA group(P0.01). Compared with the OGD group and OGD+SIRT1 siRNA group, the treatment of XFZY inhibited the decline in SIRT1 mRNA and protein expressions(P0.01), and down-regulated the mRNA and protein levels of P53, FoxO1, FoxO3, FoxO4 and NF-κB, respectively(P0.05 or P0.01). Conclusion: XFZY could prevent myocardial cells apoptosis probably by increasing the mRNA and protein expressions of SIRT1 and inhibiting the m RNA and protein expressions of P53, NF-κB, FoxO1, FoxO3 and FoxO4.     

Effects of curcumin on peroxisome proliferator-activated receptor γ expression and nuclear translocation/redistribution in culture-activated rat hepatic stellate cells

Background The function of peroxisome proliferator-activated receptor γ （PPARγ） in hepatic fibrogenesis remains largely unknown. Curcumin is a natural substance extracted form Curcuma Longa Linn and has a variety of pharmacological effects. In this study, the effects of curcumin on the proliferation, activation and apoptosis of rat hepatic stellate cells （HSCs） through PPARγ signaling were investigated. Methods HSCs were isolated from the normal Sprague Dawley rats through in situ peffusion of the liver with Pronase E and density-gradient centrifugation with Nycodenz. Cells were treated with curcumin, troglitazone, salvianolic acid B or GW9662. The effect on HSCs proliferation was determined by MTT colorimetry. Total RNA was extracted by TRizol reagent and gene levels were determined by semi-quantitative RT-PCR. Total cellular and nuclear protein were isolated and separated by 10% sodium dodecy Isulfate polyacrylamide gel electrophoresis. Protein levels were determined by Western blot. Cell apoptosis was detected by Hoechst 33258 staining. PPARγ subcellular distribution was detected by immunofluorescent staining. The activities of MMP-2 and 9 were measured by Gelatin zymograph assay. Results Curcumin suppressed HSCs proliferation in a dose-dependent manner. As HSCs underwent gradual activation with culture prolongation the PPARγ nuclear expression level decreased. Curcumin up-regulated PPARγ expression and significantly inhibited the production of a-SMA and collagen Ⅰ. PPARγ is expressed in the cytoplasm and nucleus and is evenly distributed in HSCs, but accumulated in the nucleus of HSCs and disappeared from cytoplasm after curcumin treatment. Hoechst 33258 staining showed that curcumin induced the apoptosis of culture-activated HSCs and significantly increased pro-apoptotic Bax expression and reduced anti-apoptotic Bcl-2 expression. Cyclin D1 gene, activated NFKB p65 protein and TGFβR-Ⅰ protein expression were down-regulated significantly by curcumin. The activities of MMP-2 and MMP-9 were enhanced significantly by curcumin. Conclusions Curcumin can inhibit the proliferation and activation of HSCs, induce the apoptosis of activated HSCs and enhance the activities of MMP-2 and MMP-9. The effects of curcumin are mediated through activating the PPARγ sianal transduction pathway and associated with PPARγ nuclear translocation/redistribution.     

Evaluation of oxidative stress in colorectal cancer patients

Objective To evaluate the oxidative stress in patients with colorectal cancer and to investigate the relationship between oxidative stress and colorectal cancer. Methods Seventy-six subjects were divided into two groups （36 colorectal cancer patients as the study group and 40 normal healthy individuals as the control group）. Their protein oxidation, DNA damage, lipid peroxidation and antioxidants, vitamin C, vitamin E, glutathione （GSH）, and antioxidative enzymes in serum were detected. Results The levels of protein carbonyl and advanced oxidation protein products （AOPP） were significantly higher in the study group than in the control group （P〈0.01）. Serum 8-OHdG was significantly increased in the study group compared to the control group （P〈0.01）. However, the mean serum level of MDA and conjugated diene was lower in the study group than in the control group （P〈0.01）. The activity of antioxidative enzymes was significantly decreased in the study group compared to the control group （P〈0.01）. Serum vitamins C and E concentrations were significantly reduced in the study group compared to the control group （P〈0.01）. Conclusion Colorectal cancer is associated with oxidative stress, and assessment of oxidative stress and given antioxidants is important for the treatment and prevention of colorectal cancer.     

Guizhi Decoction (桂枝汤) Inhibits Cholinergic Transdifferentiation by Regulating Imbalance of NGF and LIF in Salt-Sensitive Hypertensive Heart Failure Rats

Objective: To observe the imbalance of anatomical and functional innervation factors of sympathetic nerves, nerve growth factor(NGF) and leukemia inhibitory factor(LIF), in salt-sensitive hypertensive heart failure rats and to explore the effects of treatment with Guizhi Decoction(桂枝汤) on sympathetic remodeling by inhibiting cholinergic transdifferentiation. Methods: SS-13 BN and Dahl salt-sensitive(DS) rats were divided into 3 groups: SS-13 BN group(control group, n=9), DS group(model group, n=9) and GS group(Guizhi Decoction, n=9). After 10 weeks of a high-salt diet, the GS group rats were given Guizhi Decoction and other two groups were given saline at an equal volume as a vehicle. After 4 weeks' intragastric administration, rats were executed to detect the relevant indicators. Echocardiography and plasma n-terminal pro-B type natriuretic peptide(NT-proBNP) levels were used to assess cardiac function. Noradrenaline(NA) levels in the plasma and myocardium were detected to evaluate the sympathetic function. NGF and LIF expression were detected in the myocardium by Western blot or quantitative real-time PCR. Double immunofluorescence or Western blot was used to detect tyrosine hydroxylase(TH), choline acetyltransferase(CHAT) and growth associated protein 43(GAP43) in order to reflect anatomical and functional changes of sympathetic nerves. Results: DS group had anatomical and functional deterioration of sympathetic nerves in the decompensation period of heart failure compared with SS-13 BN group. Compared with the DS group, Guizhi Decoction significantly decreased the expression of LIF mRNA/protein(P0.01), increased the expression of NGF(P0.05 or P0.01), enhanced the levels of TH~+/GAP43~+ and TH~+/CHAT~+ positive nerve fibers(P0.01), and improved the protein expression of TH and GAP43 in left ventricle, but had no effect on CHAT(P0.05). Guizhi Decoction inhibited inflammatory infiltration and collagen deposition of myocardial injury, increased the content of myocardial NA(P0.05), reduced the plasma NA level(P0.01), improved cardiac function(P0.01), and improved weight and blood pressure to some extent(P0.05), compared with DS group. Conclusions: Guizhi Decoction could inhibit cholinergic transdifferentiation of sympathetic nerves, improve the anatomical and functional denervation of sympathetic nerves, and delay the progression of decompensated heart failure. The mechanism may be associated with the correction of the imbalance of NGF and LIF.