Fuzheng Huayu Formula (扶正化瘀方) Prevents Rat Renal Interstitial Fibrosis Induced by HgCl2 via Antioxidative Stress and Down-regulation of Nuclear Factor-Kappa B Activity

Objective: To investigate the mechanism of action of Fuzheng Huayu Formula (扶正化瘀方, FZHY) against renal interstitial fibrosis (RIF) relating to oxidative injury and nuclear factor-kappa B (NF-κB) activity. Methods: Thirty-two Sprague-Dawley rats were randomly divided into 3 groups: normal group, model group and FZHY treatment group. The RIF model was induced by oral administration of HgCl2 at a dose of 8 mg/kg body weight once a day for 9 weeks. Meanwhile, rats in FZHY treatment group orally took FZHY at a dose of 4.0 g/kg rat weight for 9 weeks. The content of hydroxyproline (Hyp) and collagen deposition in kidney were observed. The activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), the content of glutathione (GSH) and malondialdehyde (MDA) of kidney were tested. The expressions of inhibitor-κappa B (IκB), phospho-IκB (p-IκB), tumor necrosis factor-α (TNF-α), matrix metalloproteinase-2 (MMP-2) and α-smooth muscle actin (α-SMA) were analyzed by Western blot. α-SMA expression was also observed by immunofluorescent staining. MMP-2 activity was measured by gelatin zymography. NF-κB activation was determined by electrophoretic mobility shift assay. Results: Renal interstitial fibrosis was induced by HgCl2, demonstrated by remarkably increased Hyp contents and excessive collagen deposition in kidney (P<0.01). FZHY significantly inhibited renal interstitial collagen deposition and reduced Hyp content of the HgCl2-treated rats (P<0.01). GSH content decreased obviously, and MDA content increased significantly in HgCl2-treated rats compared with that of normal rats (P<0.01). FZHY significantly increased GSH content and decreased MDA content in the model rats (P<0.01). The expression α-SMA was increased in model rats compared with that of normal rats, FZHY significantly decreased its expression (P<0.01). The expressions of p-IκB and TNF-α and MMP-2, MMP-2 activity, and NF-κB activation were increased in model group compared with that in normal group (P<0.01), FZHY significantly decreased NF-κB activation, MMP-2 activity and p-IκB and TNF-α expressions (P<0.01). Conclusions: FZHY could protect kidney from oxidative injury intoxicated by HgCl2, and antagonized oxidative stress-stimulated NF-κB activity through inhibition of IκB phosphorylation in the interstitial fibrotic kidney, these effects importantly contributed to FZHY action mechanism against renal interstitial fibrosis.     

Fuzheng Huayu Formula(扶正化瘀方) Prevents Rat Renal Interstitial Fibrosis Induced by HgCl_2 via Antioxidative Stress and Down-regulation of Nuclear Factor-Kappa B Activity

Objective:To investigate the mechanism of action of Fuzheng Huayu Formula(扶正化瘀方,FZHY)against renal interstitial fibrosis(RIF)relating to oxidative injury and nuclear factor-kappa B(NF-κB)activity.Methods:Thirty-two Sprague-Dawley rats were randomly divided into 3 groups:normal group,model group and FZHY treatment group.The RIF model was induced by oral administration of HgC l2 at a dose of 8 mg/kg body weight once a day for 9 weeks.Meanwhile,rats in FZHY treatment group orally took FZHY at a dose of4.0 g/kg rat weight for 9 weeks.The content of hydroxyproline(Hyp)and collagen deposition in kidney were observed.The activities of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px),the content of glutathione(GSH)and malondialdehyde(MDA)of kidney were tested.The expressions of inhibitor-κappa B(IκB),phospho-IκB(p-IκB),tumor necrosis factor-α(TNF-α),matrix metalloproteinase-2(MMP-2)andα-smooth muscle actin(α-SMA)were analyzed by Western blot.α-SMA expression was also observed by immunofluorescent staining.MMP-2 activity was measured by gelatin zymography.NF-κB activation was determined by electrophoretic mobility shift assay.Results:Renal interstitial fibrosis was induced by Hg Cl2,demonstrated by remarkably increased Hyp contents and excessive collagen deposition in kidney(P0.01).FZHY significantly inhibited renal interstitial collagen deposition and reduced Hyp content of the Hg Cl2-treated rats(P0.01).GSH content decreased obviously,and MDA content increased significantly in HgC l2-treated rats compared with that of normal rats(P0.01).FZHY significantly increased GSH content and decreased MDA content in the model rats(P0.01).The expressionα-SMA was increased in model rats compared with that of normal rats,FZHY significantly decreased its expression(P0.01).The expressions of p-IκB and TNF-αand MMP-2,MMP-2 activity,and NF-κB activation were increased in model group compared with that in normal group(P0.01),FZHY significantly decreased NF-κB activation,MMP-2 activity and p-IκB and TNF-αexpressions(P0.01).Conclusions:FZHY could protect kidney from oxidative injury intoxicated by Hg Cl2,and antagonized oxidative stress-stimulated NF-κB activity through inhibition of IκB phosphorylation in the interstitial fibrotic kidney,these effects importantly contributed to FZHY action mechanism against renal interstitial fibrosis.     

The role of angiopoietin-1 and thrombospondin-1 in the kidney of rats subject to 5/6 nephrectomy

The expression of angiopoietin- 1 （Ang- 1） and thrombospondin- 1 （TSP- 1） in 5/6 subtotal nephrectomy （STN） rats model, and its correlation to the renal microvasculature injury were investigated. Rat 5/6 STN model was established in adult male SD rats, and the sham-operated group and 5/6 STN group were set up. The renal function and histopathological changes were examined at the 1st, 2nd, 4th, 8th and 12th week after operation. The expression orAng-1, TSP-1 and CD31 in renal tissues was detected by using immunohistochemistry. From 2nd to 8th week after operation, Ang-1 was significantly expressed in glomeruli of rats with STN. Ang-1 staining in glomeruli of STN group was increased significantly as compared with that in sham-operated group at 4th and 8th week after operation, and subsequently decreased after the 12th week. The expression of TSP-1 was increased significantly in STN group. As compared with sham-operated group, the CD31 expression was significantly down-regulated from the 2nd week. The expression of Ang-1 mRNA was detected by using RT-PCR at the same time points. The expression of Ang-1 mRNA in renal tissue of rats with STN was significantly up-regulated at the 2nd, 4th and 8th week after operation as compared with that in STN group at other time points or in sham-operated group at the same time points, while decreased evidently at the 12th week as compared with that in sham-operated group. It is concluded that there are changes in the mRNA expression of Ang-1, and the significant up-regulation of the expression of TSP-1 in renal tissue of rats with STN, which may be involved in the remnant renal microvasculature injury.     

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.     

Role of BMP-7 and TGF-β1 expression in renal tubulo-interstitial lesion of Wistar rats induced by unilateral ureteral obstruction

Objective： To explore the role of bone morphorgenetic protein-7 （BMP-7） in the renal tubulo-interstitial lesions induced by unilateral ureteral obstruction （UUO）. Methods： Sixty Wistar rats were equally and randomly divided into normal control, sham operation and UUO groups, and respectively sacrificed on the 1st, 3rd, 7th, 14th day after the time of UUO operation. The mRNA levels of BMP-7 and TGF-β1 in the renal tissues were examined by RT-PCR. The expression sites and levels of BMP-7 and TGF-β1 proteins were detected by immunohistochemistry staining. Results：Compared to control groups, the level of BMP-7 mRNA was significantly decreased, but that of TGF-β1 mRNA was significantly increased in UUO rats. Immunohistochemistry staining indicated that BMP-7 mainly expressed in the renal tubules and interstitum, rarely in the glomeruli. In UUO rats, the expression of BMP-7 protein was decreased, but that of TGF-β1 was increased in an obstruction dependent manner. Conclusion：The downregulation of BMP-7 is observed in the early phase of fibrotic process of the renal interstitium, suggesting it may be involved in the formation and development of the tubulo-interstitial lesions.     

Effects of <Emphasis Type="Italic">Panax notoginoside</Emphasis> on the expression of TGF-β1 and Smad-7 in renal tissues of diabetic rats

In order to explore the effects of Panax notoginoside (PNS) on the expression of transforming growth factor β1 (TGF-β1) and Smad-7 in renal tissues of diabetes, a rat model of diabetic nephropathy was set up by intravenous injection of streptozotocin (STZ). Wistar rats were randomly divided into normal group, diabetic control group, group treated by PNS at low-dosage (PL), group treated by PNS at high-dosage (PH) and group treated by catopril (C), respectively. Fasting blood glucose (FBG), renal index, endogenous creatinine clearance rate (CCr) and urinary albumin (UAlb) in 24 h were examined after 6 weeks. Meanwhile, the expressions of TGF-β1 and Smad7 in renal tissues were immunohistochemically dectected. At the end of the sixth week, FBG, renal index, Ccr, UAlb were all elevated significantly in control group (P<0.01). The expression of TGF-β1 protein was increased while Smad7 protein decreased in renal tissue (P<0.01). However, the treatment with PNS reversed the aforementioned changes in renal tissues of diabetic rats. These results indicate that PNS possess a protective effect on the kidney of diabetic rats and it might protect kidney by inhibiting the expression of TGF-β1 protein and enhancing the expression of Smad7 protein.     

Fasudil hydrochloride hydrate, a Rho-kinase inhibitor, ameliorates hepatic fibrosis in rats with type 2 diabetes

Background Hyperglycemia may accelerate liver fibrosis.Currently,there is no effective treatment for liver fibrosis induced by type 2 diabetes.The study aim was to investigate whether RhoA/Rho kinase （ROCK） pathway is involved in liver fibrosis in the rats with type 2 diabetes and define the protective effects of fasudil on livers.Methods A rat model of type 2 diabetes was established by high fat diet combined with streptozotocin （30 mg/kg,intraperitoneal injection）.Animals were randomly assigned to 3 groups：control rats,untreated diabetic rats that received vehicle and fasudil-treated diabetic rats that received ROCK inhibitor fasudil hydrochloride hydrate （10 mg/kg per day,intraperitoneal injection,for 14 weeks）.The morphological features of liver were observed by HE staining.Accumulation of collagen in livers was determined by Masson staining and the measurement of hydroxyproline.The mRNA expression of transforming growth factor-β1 （TGFβ1）,connective tissue growth factor （CTGF）,type-Ⅰ,and type-Ⅲ procollagen was assessed with real-time polymerase chain reaction.The phosphorylation of myosin phosphatase target subunit-1 （MYPT1）and the protein levels of TGFβ1 and α-smooth muscle actin （α-SMA） were evaluated by Western blotting.Results Compared with control rats,untreated diabetic rats showed higher values of collagen and hydroxyproline in livers （P ＜0.01）,the phosphorylation of MYPT1 and the protein levels of TGFβ1 and α-SMA were increased （P ＜0.01）,and the mRNA expression of TGFβ1,CTGF,type-Ⅰ,and type-Ⅲ procollagen was upregulated （P ＜0.01）; compared with untreated diabetic rats,treatment with fasudil signifcantly reduced values of collagen and hydroxyproline （P ＜0.01）,and decreased the phosphorylation of MYPT1 and the levels of TGFβ1 and α-SMA （P ＜0.01）,concomitant with the downregulation of TGFβ1/CTGF,type-Ⅰ,and type-Ⅲ procollagen mRNA expression （P ＜0.01）.Conclusions Fasudil ameliorates liver fibrosis in rats with type 2 diabetes at least partly by inhibiting TGFβ1/CTGF pathway and α-SMA expression.Inhibition of RhoA/ROCK may be a novel therapeutic target for liver fibrosis in diabetic non-alcoholic steatohepatitis.     

Effects of Houttuynia Cordata Thumb on Expression of BMP-7 and TGF-β1 in the Renal Tissues of Diabetic Rats

Objective： To explore the effects of Houttuynia Cordata Thumb （HCT 鱼腥草 Yu Xing Cao） on expression of transforming growth factor-β1 （TGF-β1） and bone morphogenetic protein-7 （BMP-7） in the renal tissues of diabetic rats. Methods： The diabetic rats induced by intravenous injection of streptozotocin（STZ） were randomly divided into a model group, a HCT group and a lotensin group, with normal rats designated as the controls. 8 weeks later, the ratio of kidney weight to body weight, the glomerular area, the excretion of β 2-microglobin （β2-MG） in 24-hr urine, the albumin excretion in 24-hr urine, and creatinine clearance rate （CCR） were investigated. The expression of TGF- β 1, BMP-7 and collagen I in the renal tissues was observed with the immunohistochemical method and by the semi-quantitative assay. Results： The overgrowth of glomerulus, the excretion of β 2-MG in 24-hr urine, the albumin excretion rate in 24-hr urine and CCR in the HCT group significantly reduced （P〈0.05）, and the expression of TGF-β1 and collagen I significantly decreased （P〈0.05）, but BMP-7 significantly increased （P〈0.05） in the HCT group as compared with those in the model group, with no significant difference as compared with the lotensin group （P〉0.05）. Conclusion： HCT has a protective effect on the renal tissues in diabetic rats, which is probably correlated with the decrease of the expression of TGF-β1 and collagen I and with the increase of the expression of BMP-7 in the renal tissues.     

Effect of Yishen Huayu Fang on kidney tissue E-cadherin expression in unilateral ureter ligation in rats

OBJECTIVE:To observe E-calcium sticky protein(E-cadherin) expression in kidney tissues in a rat model of unilateral ureter ligation and the effect of Yishen Huayu Fang(formula of tonifying the kidney and dissolving accumulated blood stasis) on the expression.METHODS:A total of 150 clean grade male rats were randomly divided into a control group,model group,low-dose Yishen Huayu Fang group(low-dose group),high-dose Yishen Huayu Fang group(high-dose group),and Lotensin group.A renal fibrosis model was established with unilateral ureteral obstruction(UUO).Pathological changes of rat renal tissue were observed with light microscopy on days 3,7,14,21,and 28 after UUO.Changes in kidney tissue E-cadherin expression were observed with immunohistochemistry.RESULTS:Three days after modeling,kidney edema appeared followed by gradual inflammatory cell infiltration,and part of the small tubules disappeared while the renal cortex thinned.Meanwhile,the E-cadherin expression level dropped,which was negatively correlated with the obstruction time.After intervention,E-cadherin expression was increased in all treatment groups(P<0.01 or P<0.05),while there were no significant differences between the high-dose and Lotensin groups.CONCLUSION:Yishen Huayu Fang delays the renal fibrosis process by promoting E-cadherin expression in renal tissues and reducing extracellular matrix deposition.     

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.     

Expression of BMP-7 and its receptors in renal tubolo-interstitial fibrosis induced with unilateral ureteral obstruction in rats

Objective:To study the changes of the expression of bone morphogenetic protein-7(BMP-7) and its receptors(BMPR- Ⅰ ,ALK2,ALK3 and ALK6) in the renal tubulointerstitial fibrosis induced with unilateral ureteral obstruction in rats. Methods: Sixty Wistar male rats were divided randomly into the normal control,sham-operation and unilateral ureteral obstruction(UUO) groups and the rats were killedon the 1^st , 3^rd, 7^th and 14^th postoperative days respectively. The mRNA level of BMP-7, BMPR- Ⅰ , ALK2, ALK3 and ALK6 was determined with RT-PCR. The site and level of protein expression of BMP-7 were observed with immunohistochemical staining. Results: The mRNA level of BMP-7, BMPR- Ⅰ ,ALK2 and ALK3 was significantly decreased in the rats of UUO group than in those of the sham-operation group but the mRNA level of ALK6 showed no obvious changes in all the rats. Immunohistochemical staining revealed that the protein of BMP-7 was mainly expressed in the renal tubules and interstitial tissue of the kidneys in normal rats but it was decreased gradually along with the unilateral ureteral obstruction. Conclusion: It is found that the loss of BMP-7 and its receptors including BMPR- Ⅰ ,ALK2 and ALK3 occursin the early phase of renal tubulointerstitial fibrosis before the appearance of other pathological changes in the kidney and may play an important role in the occurrence and progress of renal tubulointerstitial fibrosis.     

Effect of expression of TGF-beta and TNF-alpha with Qidan granule treatment in rats by bleomycin-induced pulmonary fibrosis

Objective: To evaluate the therapeutic effects and mechanisms of Qidan granule in blemycinA5-induced pulmonary interstitial fibrosis (PIF)in rats. Methods: PIF models were established by blemycinA5-induced in rats. They were treated by Qidan granule and Hydrocortisone respectively. The pathological changes and collagen protein disposition were observed, and the expression of TGF-β, TNF-α proteins were measured by immunohistochemical technique . Results: The pulmonary alveolitis and fibrosis were alleviated remarkably in Qidan granule group compared with those in the model control group and hydrocortisone group (P <0. 01). The expression of TGF-β and TNF-α protein were higher in Qidan granule group than those in normal group , and were significantly less than those in the model control group and in hydrocortisone group (P < 0. 01). Conclusion: Qidan granule would ameliorate the pulmonary alveolitis and fibrosis. TGF-β and TNF-α might play an important role in the development of alveolitis and fibro     

Preliminary Study on the Mechanism of Acupoint Injection of Bone Marrow Mesenchymal Stem Cells in Improving Blood Flow in the Rat of Hind Limb Ischemia

Objective:To explore the mechanism of acupoint injection of bone marrow mesenchymal stem cells(BM-MSCs) in improving blood flow in the rat with hind limb ischemia.Methods:Twenty-four SD rats were randomly divided into 4 groups:normal control group(n=6),model group(n=6),BM-MSCs acupoint injection group(AI group,n=6) and BM-MSC intramuscular injection group(MI group,n=6).Sanyinjiao(SP 6),Housanli(ST 36),Zhaohai(KI 6),Huantiao(GB 30) and Yanglingquan(GB 34) were selected for the AI group,and five non-acupoints were selected on gastrocnemius and adductor of ischemic hind limbs in the MI group.BM-MSCs were injected to the latter two groups.The rat hind limb ischemia model was established with the method of blocking the femoral artery and its branches.Three weeks after injection of BM-MSCs,in each group,hindlimb adductor and gastrocnemius were taken from the ischemic side.Expressions of vascular endothelial growth factor(VEGF) and transfer growth factor-β1(TGF-β1) in the skeletal muscle were determined with immunohistochemical method,and the small arteries in the skeletal muscle were labeled with α-SMA immunohistochemical staining method,the density of small arteries(number of arterioles /number of muscle fibers) and the number of the blood vessel with VEGF positive expression were calculated.The serum levels of VEGF and nitric oxide(NO) were detected.Results:Compared with the model group,the expression of VEGF and TGF-β1,and the density of small arteries and the number of VEGF-positive blood vessels in the AI group and the MI group significantly increased(both P<0.01).Compared with the MI group,the density of small arteries and the number of VEGF-positive blood vessels in the AI group significantly increased(both P<0.01);Compared with the model group and the normal control group,the serum expression quantity of NO and VEGF in the AI group and the MI group were significantly increased(P<0.01).Conclusions:Acuppoint injection of BM-MSCs secrets more VEGF,TGF-β1 and NO to increase angiogenesis and arteriogenesis,so as to improve blood flow of the rats of hind limb ischemic.     

Salvianolate Reduces Glucose Metabolism Disorders in Dimethylnitrosamine-Induced Cirrhotic Rats

Objective: To evaluate the preventive effect of salvianolate(Sal B) on glucose metabolism disorders of dimethylnitrosamine(DMN)-induced cirrhotic rats. Methods: Fifty-five Wistar rats were randomly divided into a control group(n=10) and a cirrhotic group(n=45) according to a random number table. Liver cirrhosis was induced by intraperitoneal administration of DMN. The cirrhotic rats were divided into model, Sal B and metformin groups(n=15), respectively. Rats in the model group were given saline, two treatment groups were given Sal B(50 mg/kg), metformin(150 mg/kg) respectively for 28 consecutive days, while rats in the control group were injected 0.9% saline with same volume of vehicle. Body weight was measured everyday. Insulin sensitivity was determined by euglycemic hyperinsulinemic clamp. Organ index, glucose tolerance test(OGTT), and fasting plasma glucose(FPG), fasting insulin(FINS), hepatic glycogen, hydroxyproline(HYP) and liver function were detected at the end of the treatment. Area under the curve(AUC) for OGTT was calculated. Liver and pancreas histology were determined by histopathological examination with hematoxylin and eosin staining(HE), Sirius Red staining and Masson's trichrome staining, respectively. Hepatic expression of α-smooth muscle actin(α-SMA) and collagen(Col Ⅰ) were evaluated by immunohistochemical staining. Results: Compared with the model group, Sal B significantly increased body and liver weight, liver-body ratio, glucose infusion rate(GIR), FPG, FINS levels and hepatic glycogen at the end of administration(P0.05 or P0.01). Meanwhile, Sal B significantly decreased AUC for OGTT, spleen weight, spleen-body ratio, aminotransferase and HYP level(P0.05 or P0.01). Sal B was also effective in alleviating necrosis of liver tissue, suppressing fibrosis progression and inhibiting the expression of α-SMA and Col Ⅰ in liver. Compared with the metformin group, Sal B had advantages in ameliorating FPG, hepatic glycogen, spleen weight, organ index, liver function and cirrhosis(P0.05). Metformin increased insulin sensitivity more potently than Sal B(P0.05). Conclusions: Sal B could improve glucose metabolism in cirrhotic rats by protecting hepatic glycogen reserve, increasing insulin sensitivity, and alleviating pancreatic morphology abnormalities. Sal B was clinically potential in preventing glucose metabolism anomalies accompanied with cirrhosis.     

桂枝汤对糖尿病大鼠心率变异性及心脏自主神经紊乱的调节作用

Objective:To observe abnormalities in heart rate variability(HRV) in diabetic rats and to explore the effects of treatment with Guizhi Decoction(桂枝汤) on cardiac autonomic nervous(CAN)imbalance.Methods:A radio-telemetry system for monitoring physiological parameters was implanted into rats to record electrocardiac signals and all indictors of HRV[time domain measures:standard deviation of all RR intervals in 24 h(SDNN),root mean square of successive differences(RMSSD),percentage of differences between adjacent RR intervals greater than 50 ms(PNN50),and standard deviation of the averages of RR intervals(SDANN);frequency domain measures:low frequency(LF),high frequency(HF),total power(TP),and LF/HF ratio].The normal group was randomly selected,and the remaining rats were used to establish streptozocin(STZ)-induced diabetic model.After 4 weeks,the model rats were divided into the model group,the methycobal group,and the Guizhi Decoction group,9 rats in each group.Four weeks after intragastric administration of the corresponding drugs,the right atria of the rats were collected for immunohistochemical staining of tyrosine hydroxylase(TH) and choline acetyltransferase(CHAT) to observe the distribution of the sympathetic and vagus nerves in the right atrium.The myocardial homogenate from the interventricular septum and the left ventricle was used for determination of TH,CHAT,growth-associated protein 43(GAP-43),nerve growth factor(NGF),and ciliary neurotrophic factor(CNTF) levels using an enzyme-linked immunosorbent assay.Results:(1) STZ rats had elevated blood glucose levels,reduced body weight,and decreased heart rate;there was no difference between the model group and the drug treated groups.(2) Compared with the model group,only RMSSD and TP increased in the methycobal group significantly(P0.05);SDNN,RMSSD,PNN50,LF,HF,and TP increased,LF/HF decreased(P0.05),and SDANN just showed a decreasing trend in the Guizhi Decoction group(P0.05).TH increased,CHAT decreased,and TH/CHAT increased in the myocardial homogenate of the model group(P0.05).Compared with the model group,left ventricular TH reduced in the methycobal group;and in the Guizhi Decoction group CHAT increased,while TH and TH/CHAT decreased(P0.05).Compared with the model group,CNTF in the interventricular septum increased in the methycobal group(P0.05);GAP-43 increased,NGF decreased,and CNTF increased(P0.05) in the Guizhi Decoction group.There were significant differences in the reduction of NGF and elevation of CNTF between the Guizhi Decoction group and the methycobal group(P0.05).(3) Immunohistochemical results showed that TH expression significantly increased and CHAT expression significantly decreased in the myocardia of the model group,whereas TH expression decreased and CHAT expression increased in the Guizhi Decoction group(P0.05).Conclusion:Guizhi Decoction was effective in improving the function of the vagus nerve,and it could alleviate autonomic nerve damage.