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BACKGROUND: The relationship between nuclear factor (NF)-kB signaling pathway and intervertebral disc degeneration is a hotspot in the field of orthopedics. In-depth investigation on various signaling pathways in the intervertebral disc contributes to understanding the mechanism of intervertebral disc degeneration. OBJECTIVE: To investigate the regulation of the serum containing Yishen Huoxue Tongluo Recipe on the NF-kB signal pathway of human intervertebral disc nucleus pulposus cells under different hydrostatic pressures, attempting to explore the possible therapeutic mechanism and target of Yishen Huoxue Tongluo Recipe in the treatment of degenerative diseases of the intervertebral disc from the perspective of molecular biology. METHODS: Passage 3 Human intervertebral disc nucleus pulposus cells were divided into eight groups and were cultured in the drug-containing serum of Yishen Huoxue Tongluo Recipe. The cells were intervened for 2, 4, and 6 hours under different hydrostatic loading conditions (0.3,1, and 3 MPa). The morphology and growth of nucleus pulposus cells were observed by an inverted phase contrast microscope. Ultrastructural changes of intervertebral disc nucleus pulposus cells were observed by a transmission electron microscopy. Cell counting kit-8 method was used to detect the proliferation activity of nucleus pulposus cells. Annexin V-FITC/Propidium Iodide apoptotic kit was used to double-stain nucleus pulposus cells to detect the cell apoptotic rate. Western blot method was used to detect the changes of NF-kB p65, Collagen II, ADAMTS-4, MMP-13 and Caspase-3 in nucleus pulposus cells. RESULTS AND CONCLUSION: (1) Under the same pressure and time, the morphology and growth of nucleus pulposus cells in the pressure+drug-containing serum groups were better than those in the normal pressure group and the simple pressure groups. Among them, the 0.3 and 1 MPa pressure+drug-containing serum groups had more intact cell morphology and better cell growth than the 3 MPa pressure+ drug-containing serum group. (2) The proliferative activity of nucleus pulposus cells was higher in the pressure+drug-containing serum group, and there was a significant difference between the 0.3 MPa pressure+drug-containing serum group and the 0.3 MPa simple pressure group (P < 0.05). (3) The apoptotic rate of nucleus pulposus cells in the pressure+drug-containing serum group was lower than that in the normal pressure group and simple pressure intervention group (P < 0.05). (4) The expression of Collagen II and Caspase-3 increased in the pressure+drug-containing serum group, while the expression of NF-kappa B p65, ADAMTS-4 and MMP-13 decreased in the pressure+drug-containing serum group (P < 0.05). To conclude, Yishen Huoxue Tongluo Recipe can increase cell activity, reduce cell apoptosis and effectively delay the degeneration of nucleus pulposus cells. Its mechanism is likely to promote the expression of Collagen II and Caspase-3 through the NF-kB signaling pathway of nucleus pulposus cells of intervertebral disc, and inhibit the expression of NF-kB p65, ADAMTS-4 and MMP-13.
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Objective@#To explore the protective effect of Yishen Tongluo Recipe (YTR) against aberrant sperm DNA methylation in male rats exposed to benzo(a)pyrene (BaP).@*METHODS@#Thirty male SD rats of the SPF grade were randomly divided into three groups of equal number: solvent control, BaP exposure and YTR intervention. The animals of the solvent control group were injected intraperitoneally with 0.5% DMSO while those of the other two groups with BaP at 0.1 mg/kg/d, all for 60 days, and at 31 days of BaP exposure, those of the YTR group were treated intragastrically with YTR for 30 days. Then, the left epididymides were harvested from all the rats and sperm suspensions collected and centrifuged for extraction of sperm DNA. The methylated DNA immunoprecipitation sequencing (MeDIP-seq) technique was used to detect the whole-genome DNA methylation in different groups.@*RESULTS@#Exposure to BaP induced the up-regulation of 828 genes encoding mRNA in the sperm DNA, while YTR intervention produced a significant protective effect on the transforming growth factor β3 (TGF-β3), cystic fibrosis transmembrane conductance regulator (CFTR) and recombination activating gene 1 (RAG1), and down-regulated the expressions of 3 227 genes. BaP exposure also caused the up-regulation of 783 genes encoding lncRNA in the sperm DNA, and YTR treatment exhibited an evident protective effect on 62 of the up-regulated genes, induced the down-regulation of 3 378 genes, and showed a protective effect on 56 of the down-regulated genes.@*CONCLUSIONS@#YTR has a protective effect against aberrant sperm DNA methylation in male rats exposed to BaP, which may be associated with lncRNA.
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Objective To study the clinical efficacy of Bushen Huoxue Tongluo Recipe in the treatment of type 2 diabetes with hypertension, to provide more basis for clinical treatment.Methods64 patients with type 2 diabetes mellitus complicated with hypertension were selected in our hospital from July 2014 to November 2015.They were divided into two groups, each group was 32.The control group was treated with western medicine, the observation group was treated with Bushen Huoxue Tongluo recipe, and the therapeutic effects of the two groups were observed and compared.ResultsThe total effective rate of the two groups has significantly difference, and the incidence of complications of the two groups also has statistical difference (P<0.05).ConclusionThe treatment of Bushen Huoxue Tongluo recipein in type 2 diabetes with hypertension, has good effect, was very useful to change symptoms of patients, improve the clinical treatment effect of patients.It is conducive to the recovery of patient.
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Objective To investigate the clinical effect of Bushen Huoxue Tongluo Recipe in the treatment of acute attack of primary knee osteoarthritis, so as to enrich the treatment method and improve the curative effect. Methods 94 cases of primary knee osteoarthritis acute attack were randomly divided into 2 groups, 47 cases in the control group were treated with conventional western medicine treatment, the observation group of 47 cases, treated with Bushen Huoxue Tongluo Decoction, were observed after treatment in the improvement of the safety index, serum indexes and related score. Results The two groups at the end of the pain, stiffness, ADL, tenderness, swelling, joint activities are significantly lower than before treatment, after treatment were significantly different (P<0.05) at the end of treatment, the observation group in the above six indicators were significantly better than the control group (P<0.05); group MMP-3, two at the end of treatment CRP, IL-1, TNF- were significantly lower than before treatment, after treatment were significantly different (P<0.05) at the end of treatment observation group above index was significantly better than the control group (P<0.05). Conclusion: Bushen Huoxue Tongluo Decoction can improve the primary acute exacerbation of knee osteoarthritis clinical symptoms, reduce inflammation, and high safety.
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Objective To investigate the clinical effect of Bushen Huoxue Tongluo Recipe in the treatment of acute attack of primary knee osteoarthritis, so as to enrich the treatment method and improve the curative effect. Methods 94 cases of primary knee osteoarthritis acute attack were randomly divided into 2 groups, 47 cases in the control group were treated with conventional western medicine treatment, the observation group of 47 cases, treated with Bushen Huoxue Tongluo Decoction, were observed after treatment in the improvement of the safety index, serum indexes and related score. Results The two groups at the end of the pain, stiffness, ADL, tenderness, swelling, joint activities are significantly lower than before treatment, after treatment were significantly different (P<0.05) at the end of treatment, the observation group in the above six indicators were significantly better than the control group (P<0.05); group MMP-3, two at the end of treatment CRP, IL-1, TNF- were significantly lower than before treatment, after treatment were significantly different (P<0.05) at the end of treatment observation group above index was significantly better than the control group (P<0.05). Conclusion: Bushen Huoxue Tongluo Decoction can improve the primary acute exacerbation of knee osteoarthritis clinical symptoms, reduce inflammation, and high safety.
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Objective: To observe the protective effect of α-lipoic acid (ALA) and Tongluo recipe(TLR) against injury of glomerular visceral epithelial cells in diabetic rats, so as to discuss the possible mechanism for injury of glomerular visceral epithelial cells in diabetes. Methods: Rat diabetes model was induced by high-fat diet and a single intraperitoneal injection of streptozotocin (35 mg/kg). The diabetic model rats were randomly divided into three groups (n = 8): diabetes mellitus (DM) group, DM+TLR and DM+ALA group; another 8 rats served as normal controls (CON group). Rats in the DM+TLR group received 0.4 g • kg-1 • d-1 TLR and DM+ALA group received 0. 1 g • kg-1 • d-1 ALA throughout the experiment. Rats in the DM group and control (CON) group were given celiac perfusion of distilled water once at the same time and volume. After 12 weeks of treatment, kidney mass/body mass index, 24 h urinary protein, blood urea nitrogen, and urinary albumin/Cr were determined in each group. The number and the density of podocytes in glomerulis were measured by stereological dissector/ fractionator methods. The expression of nephrin in renal cortical tissue was detected by Western blotting analysis. The activity of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and the content of malondialdehyde (MDA)were measured in the plasma and renal cortical tissues. Results: After 12 weeks, the kidney mass/body mass index, 24 h urinary protein and urinary albumin/Cr in DM group were significantly increased compared to control group (P<0. 01); the number and density of podocytes in the glomerulis and nephrin expression in DM group were significantly decreased (P<0. 01). The kidney mass/ body mass index, 24 h urinary protein and urinary albumin/Cr in DM+TLR and DM+ALA groups were significantly decreased compared to DM group (P<0. 01); the number and density of podocytes and nephrin expression in DM+TLR and DM+ALA groups were significantly increased compared to DM group (P<0. 01, P<0. 05). GSH-Px and SOD activities were significantly decreased in the plasma (P<0. 01) and renal cortical tissue (P<0. 01) in DM, DM+ALA and DM+TLR groups compared to those in the CON group, and the content of MDA was significantly increased (P<0. 01, P<0. 05). GSH-Px and SOD activities were significantly increased in the plasma and renal cortical tissue (P both<0. 01) and MDA content was significantly decreased in DM+TLR and DM+ALA groups (P<0. 01, P<0. 05)compared to those in the DM group. No significant differences were found between DM + ALA group and DM + TLR group concerning the above parameters. Conclusion: TLR can ameliorate podocyte injury in diabetic rats, which might be associated with its anti-oxidative effect.
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Objective To observe the effect of Huazhuo-Jiedu-Huoxue Tongluo recipe on local inflammation of cerebral ischemia reperfusion injury in mice. Method Fifty Kunming Mice were randomly divided into five groups: model group, high-dose group of TCM, low-dose group of TCM, nimodiping group and sham operation group. Each group had 10 mice. Mice models of the right middle cerebral artery CIRI in the first four groups were established through thread embolism method. Physiological saline was given to the mice in the model group and sham operation group. Nimodipine solution was given to the mice in the Nimodipine group. High and low dose of Huazhuo-Jiedu-Huoxue-Tongluo Chinese medicine decoction were given to the mice in the high dose of TCM group and low dose of TCM group respectively after the models were successfully established. To assess the expression level of IL-6 and TNF-α by radioimmunoassay after the treatment. Results Compared with model group [(219.29±39.28) pg/ml, (1.325 ± 0.236) ng/ml] , the level of IL-6 and TNF-α of high dose of TCM group[(120.69±49.23)pg/ml, (1.086±0.178)ng/ml], low dose of TCM group[(173.97±49.03)pg/ml, (0.937 ± 0.105)ng/ml]and nimodipine group[(170.88± 47.06)pg/ml, (1.092±0.184) ng/ml]decrease (P0.05). Conclusion Huazhuo-Jiedu-Huoxue-Tongluo recipe can restrain the expression of inflammatory cytokines and reduce the infiltrating of leukocytes in mice with cerebral ischemia reperfusion injury.
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Objective: To observe the effect of Tongluo recipe(TLR) on CD4+ CD25+ regulatory T cell in the thymus of experimental diabetic rats. Methods: Totally 112 SD rats were randomly divided into three groups: control group, diabetes mellitus (DM) group, and TLR treatment group. Rat diabetic model was induced by intraperitoneal injection of streptozotocin (60 mg/kg). The TLR treatment group was further divided into two subgroups: prevention group was given intragastric TLR (0.4 g·kg-1·d-1) for 12 weeks starting from the day of streptozotocin injection (TL-1 group), and the treatment group was given intragastric TLR (0.4 g· kg -1·d-1) for 12 weeks starting from 12 weeks after streptozotocin injection (TL-2). The animals were sacrificed at different time points (The control and DM group: 4, 8, 12, 16, 20, and 24 weeks; TLR group: 12 and 24 weeks after administration of TLR). Flow cytometry, H-E staining, and immunochemistry were performed to investigate the change of CD4+ CD25+ regulatory T cell ratio in CD4+ T cells, the thymus morphology, and the expression of Foxp3 protein in the thymus. Results: Compared with the normal control group, diabetic group had a gradually shrunk thymus, a decreased ratio of CD4+ CD25+ regulatory T cells in CD4+ T cells, and a decreased expression of Foxp3. The ratios of CD4+ CD25+ regulatory T cells and the thymus indices in the two TLR groups were significantly higher than those in the diabetic model group(P<0.05). Conclusion: Tongluo recipe can greatly increase the ratio of CD4+ CD25+ regulatory T cells, thymus index, and Foxp3 expression in the thymus, and may protect the thymus CD4+ CD25 + regulatory T cells in experimental diabetic rats.
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Objective To study the effects of Tongluo recipe on p38 mitogen-activated protrin kinase (p38 MAPK)- fibronectin(FN) signal pathway in the kidney of streptozotocin-induced diabetic rats, so as to explore the possible mechanism by which Toogluo recipe protects the kidney. Methods Male Sprague-Dawley(SD) rats were randomly divided into 3 groups(n = 7): control group (CON group), model group(DM group)and Toogluo recipe group(DM+TLR group). Rats in the DM group and DM+TLR groups were administered with streptozotocin (65 mg/kg) by intraperitoneal injection to induce diabetic model. Rats in DM + TLR group were also given 1g/(kg·d) of Toogluo recipe orally after successfully establishment of diabetic model, and rats in DM and CON groups were only given equivalent volume of ddH2O. The interference lasted for 12 weeks. Biochemical indices were obtained by automatic analyzer. Coomassie brilliant blue assay was used to measure the 24-hour urine protein. Kidney cortex was subjected to PAS staining. Western blotting analysis was used to detect the protein expression of total p38 MAPK (t-p38 MAPK), phosphorylated p38 MAPK (p-p38 MAPK), total cAMP response element-binding protein (t-CREB), phosphorylated CREB (p-CREB) and FN. Results Compared with CON group, the average left kidney mass/body mass (LKM/BM), blood glucose(BG), blood urea nitrogen(BUN), serum creatinine(Scr), 24-hour urine proteins(24 h UP) and urine protein/creatinine (UP/Ucr) were significantly increased in the DM group (P< 0.05); the average BM was significantly decreased(P<0.05); and the expression of p-p38 MAPK, p-CREB, FN was increased by 2.37, 2.21, and 2.03 folds, respectively (P< 0.05). Compared with DM group, the average LKM/BM, Scr, 24 h UP and UP/Ucr were significantly decreased (P<0.05) in DM+TLR group, and the expression of p-p38 MAPK, p-CREB, FN was significantly decreased(P< 0.05). Conclusion Tongluo recipe can protect the kidney in diabetic nephropathy rats, probably through inhibition of the p38 MAPK-FN pathway.
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Objective To study the effect of Tongluo recipe on leptin and insulin in rat model of type 1 diabetes mellitus. Methods A total of 30 adult male SD rats were equally randomized into the following 3 groups: normal control group, diabetes mellitus group and Tongluo recipe group. SD rats were administered intraperitoneally with STZ (60 mg/kg) to induce the model of type 1 diabetes mellitus. The rats in Tongluo recipe group were administered intragastrically with Tongluo recipe (1.0g·kg-1·d-1), and the other two groups were administered intragastrically with double distilled water in the same dosage. The body weight, white adipose tissue weight, fasting blood glucose, fasting insulin, total cholesterol, triglyceride, free fatty acid, and leptin levels were compared between each group 8 weeks later. The expression of leptin mRNA in white adipose tissue was examined. Results Compared with diabetes mellitus group, the body weight,fasting insulin, and leptin were significantly higher (P<0.05or0.01) in the Tongluo recipe group, the levels of fasting blood glucose, total cholesterol, and free fatty acid were significantly lower (P<0.05or0.01) in Tongluo recipe group, and the expression of leptin mRNA was significantly higher (P<0.05) in the Tongluo recipe group. Conclusion Tongluo recipe may correct the disturbance in the lipid and glucose metabolism by improving the low leptin and insulin levels in diabeticrats.
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Objective To observe the effects of Tongluo reeipe on extracellular signal-regulated protein kinase (ERK1/2) pathway in the kidneys of streptozotocin-induced rat models of type 1 diabetes mellitus, and to explore the renoprotective mechanism of Tongluo reeipe. Methods Adult male Sprague-Dawley rats were randomly divided into 2 groups: control group (A, n=10) and experimental group. Rats in the experimental group were induced by intraperitoneal injection of streptozotocin (60 mg/kg body mass), and were randomly subdivided into group B (diabetic control, n=10), group C (diabetic rats treated with low dose of Tongluo recipe,0.5g/[kg · d], n=10), group D (diabetic rats treated with medium dose of Tongluo recipe, 1.0g/[kg · d], n=10), group E (diabetic rats treated with high dose of Tongluo recipe, 2.0g/[kg · d], n=10) and group F (diabetic rats treated with losartan, 30 mg/[kg · d],n=10). At the end of the 12th week rats were sacrificed, and the serum glucose, triacylglycerol(TG), total cholesterol(Chol), serum creatinine(Scr), urea nitrogen(BUN) in blood, 24-hour urine protein(Upro24), microalbuminuria (MAU), and creatinine in urine (UCR) were examined. The kidney mass/body mass (KM/BM) and creatinine clearance rate (Ccr) were calculated. The left kídneys were collected, weighed and subjected to H-E staining and PAS staining to observe the pathological changes; serum and renal tissue angiotensin II (Ang II) levels were determined by ELISA; the expressíon of phosphorylated ERK1/2 (p-ERK1/2) was detected by immunohistochemical staining and vascular endothelial growth factor (VEGF) protein expression was determined by Western blotting analysis. Results Compared with group B, group D had a slighter mesangial proliferation under light microscope. Besides, rats in group D had significantly decreased Ccr, BUN, MAU, and Upro24 in the serum, and decreased Ang II in the serum and renal tissue, and decreased KM/BM, VEGF and p-ERK1/2 levels in renal tissue compared with rats in group B(P<0.05). The medium dose of Tongluo recipe and losartan showed similar effects in improving the renal function and inhibiting Ang II, VEGF, and p-ERK1/2 expression of diabetic rats. Conclusion The renoprotective effect of Tongluo recipe may be related to inhibition of ERK1/2 cascade pathway by decreasing the expression of Ang II and VEGF.
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Objective To observe the effects of Tongluo recipe on extracellular signal-regulated protein kinase (ERK1/2) pathway in the kidneys of streptozotocin-induced rat models of type 1 diabetes mellitus, andto explore the renoprotective mechanism of Tongluo recipe. Methods Adult male Sprague-Dawley rats were randomly divided into 2 groups: control group (A, n = 10) and experimental group. Rats in the experimental group were induced by intraperitoneal injection of streptozotocin (60 mg/kg body mass), and were randomly subdivided into group B (diabetic control, n = 10), group C (diabetic rats treated with low dose of Tongluo recipe, 0. 5 g/[kg • d], n = 10), group D (diabetic rats treated with medium dose of Tongluo recipe, 1. 0 g/[kg • d], n = 10), group E (diabetic rats treated with high dose of Tongluo recipe,2. 0 g/[kg • d], n = 10) and group F (diabetic rats treated with losartan, 30 mg/[kg • d],n=10). At the end of the 12th week rats were sacrificed, and the serum glucose, triacylglycerol(TG), total cholesterol(Choi), serum creatinine(Scr), urea nitrogen(BUN) in blood, 24-hour urine protein(Upro24), microalbuminuria (MAU), and creatinine in urine(UCR) were examined. The kidney mass/body mass (KM/BM) and creatinine clearance rate (Ccr) were calculated. The left kidneyswere collected, weighed and subjected to H-E staining and PAS staining to observe the pathological changes; serum and renal tissue angiotensin II (Ang II) levels were determined by ELISA; the expression of phosphorylated ERK1/2(p-ERK1/2) was detectedby immunohistochemical staining and vascular endothelial growth factor (VEGF) protein expression was determined by Western blotting analysis. Results Compared with group B, group D had a slighter mesangial proliferation under light microscope. Besides, rats in group D had significantly decreased Ccr, BUN, MAU, and Upro24 in the serum, and decreased Ang II in the serum and renal tissue, and decreased KM/BM, VEGF and p-ERK1/2 levels in renal tissue compared with rats in group B(P<0. 05). The medium dose of Tongluo recipe and losartan showed similar effects in improving the renal function and inhibiting Ang II, VEGF, and p-ERK1/2 expression of diabetic rats. Conclusion The renoprotective effect of Tongluo recipe may be related to inhibition of ERK1/2 cascade pathway by decreasing the expression of Ang II and VEGF.
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Objective To investigate the therapeutic effects and the possible mechanisms of Tongluo recipe(TLR) on podocyte injury in streptozotocin-induced diabetic rats. Methods Diabetes was induced by a single intraperitoneal injection of streptozotocin(60 mg/kg) dissolved in citrate buffer. The diabetic model rats were randomly divided into two groups(n=12): diabetic mellitus(DM) group and DM+TLR group. Rats in the DM+TLR group received 0. 4 g/(kg • d) TLR throughout the experiment. Rats in the DM group and CON group (n = 8) were given celiac perfusion of distilled water once at the same time and dose. After 12-week treatment, 24 h urinary protein of rats was determined; the cellular ultrastructures of podocytes were observed by transmission electronic microscope; the number and the density of podocytes in glomerulis were measured by stereo logical dissector/fractionatormethods; the expression of nephrin in renal cortical tissuewas detected by Western blotting analysis; the activity of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and the content of malondialdehyde (MDA) were measured in the plasma and renal cortical tissue. Results Compared with CON group, DM group and DM+TLR group had significantly increased ratio of kidney mass to body mass, blood glucose, and 24-h proteinuria(all P<0. 01); at the 12th week, the glomerular basement membrane in DM group became conspicuously thick, foot process fusion and microvilii of podocytes were observed, and nephrin expression, podocyte number and podocyte density in the glomerulis were all significantly decreased(P < 0. 01). The morphology of the foot process was almost normal in DM+TLR group, with the basement membrane thickened, but was slighter than that in the DM group; and nephrin expression, podocyte number and podocyte density in DM+TLR group were significantly higher than those in the DM group(P<0. 01). Compared with CON group, DM group and DM+TLR group had significantly increased MDA content, and significantly decreased SOD and GSH-Px activities in the plasma and renal cortical tissue (P<0. 01, P<0. 05). Compared with DM group, DM+TLR group hadsignificantly increased SOD and GSH-Px activities and significantly decreased MDA content in plasma and renal cortical tissue(P<0. 01, P< 0.05). Conclusion TLR can ameliorate podocyte injury probably through depleting free radical production and increasing activities of antioxidatant.
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Objective: To study the protective effect of Tongluo recipe on renal function of rats with diabetic nephropathy and its influence on the oxidative stress and antioxidative system in renal cortex. Methods: Diabetes mellitus was induced in male SD rats by intraperitoneal injection of streptozotocin (55 mg/kg). Rats with a blood glucose level ≥16.7 mmol/L after 3 days were taken as diabetic rats. Twenty-one diabetic rats were evenly randomized into 3 groups, namely, untreated diabetic control (DM group), Tongluo recipe gavaged (1g · kg-1 · d-1 for 12 weeks, TLR group), and melatonin gavaged (5 mg · kg-1 · d-1 for 12 weeks, MEL group) groups; normal rats were taken as normal control (NC group). The treatments were started 3 days after streptozotocin injection and continued for 12 weeks. Kidney cortex was collected under anesthesia for electron microscopic observation; the kidney homogenates were prepared to determine the renal anti-oxidant enzyme (SOD, GSH-Px) activities and lipid proxide (MDA) levels by chromatometry. Results: Compared with NC group, rats in DM group had a higher 24 hour-urinary protein excretion (P<0.01), while those in TLR group and MEL group were significantly lower than that of DM group (P<0.01). Electron microscopic results showed that the mesangial cells and endothelial cells in DM group were swollen and had fusion /disappearance of podocytic process, increased mesenteric matrix, uneven basal membrane, and increased mitochondria with morphological disorders; while the above changes were improved in TLR and MEL group. GSH-Px and SOD activities were lower and MDA levels were higher in renal cortex of DM group than those of NC(P<0.05); treatment with Tongluo recipe and melatonin slightly increased SOD and GSH-Px activities and decreased MDA levels, with the level in MEL group significantly lower than that of DM group (P<0.05). Conclusion: Tongluo recipe can protect the renal function of diabetic rats by lowering urinary protein excretion and improving the structure of glomerular basement membrane, but it can only slightly decrease MDA level in the renal cortex, showing an unclear anti-oxidative effect.
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Objective: To study the effect of two types of Tongluo recipe (ultrafine type and fine type) on the level of oxidative stress in kidneys of diabetic Sprague-Dawley rats. Methods: Experimental diabetes was induced by intraperitoneal injection of streptozotocin (60 mg/kg) in male Sprague-Dawley rats. One week after streptozotocin injection rats with blood glucose higher than 16.7 mmol/L were taken as diabetics. The study included the following 4 groups: normal control(NC), diabetes mellitus control(DMC), diabetics treated with ultrafine Tongluo recipe (DM+UT, 1 g · kg-1 · d-1), and diabetics treated with fine Tongluo recipe (DM+FT, 1 g · kg-1 · d-1). Four weeks after treatment, 24 h urinary protein (24 h UP) and kidney weight/ body weight (KW/BW) were detected as renal function indices; malondialdehyde(MDA) and antioxidant enzymes was examined with colorimetry; and NADPH oxidase subunits p47phox and p22phox mRNA were studied with real-time PCR. Results: Compared with the NC group, DMC group had significantly higher MDA, 24 h UP, KW/BW, and glutathione peroxidase (GSH-Px) activity, and had significantly lower activity of total superoxide dismutase (TSOD). No significant difference was found in catalase(CAT) activity between the NC group and DMC group. Compared with the 2 Tongluo Recipe groups, the DMC group had significantly lower 24 h UP, KW/BW and MDA, and significantly higher activities of TSOD and GSH-Px, and no significant difference was found in their CAT activities. The effect of ultrafine type on 24 h UP, KW/BW, MDA, TSOD and GSH-Px was significantly greater than the fine type did. The expression of p47phox mRNA of NADPH oxidase in renal cortex group was significantly lower than that in the DMC group, and the latter was significantly higher than the 2 Tongluo of the NC recipe groups, and there was no significant difference between the 2 Tongluo recipe groups. There was no significant difference in p22phox mRNA expression between all the 4 groups. Conclusion: Tongluo recipe may improve the activities of antioxidant enzymes (including TSOD and GSH-Px), decrease p47phox expression, and therefore inhibit the oxidative stress in renal cortex of diabetic rats, reducing the early kidney injury.
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Objective: To investigate the therapeutic effect of Tongluo recipe(TLR) on hepatic injury in streptozotocininduced diabetic rats and the possible mechanism. Methods: Experimental diabetes was induced by intraperitoneal injection of streptozotocin(60 mg/kg) in male Sprague-Dawley rats. The study included 5 groups: normal control(CN) group, diabetes mellitus control (DM) group, low-dose TLR (TL) group, middle-dose TLR (TM) group and high-dose TLR (TH) group, with 10 rats in each group. Diabetic rats in the latter three groups received 0.5 g/(kg·d), 1.0 g/(kg·d), 2.0 g/(kg·d) TLR, respectively throughout the experiment. The fasting glucose, blood lipid, and serum liver function indices were measured. Twelve weeks after treatment, rats were sacrificed and the liver tissues were collected to examine the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), and malondialdehyde (MDA). The ultrastructure of liver tissues was observed through transmission electron microscope. Results: The serum AST, ALT and AKP levels of diabetic rats were increased 12 weeks after intraperitoneal injection of streptozotocin (P<0.01), and electron microscope observation showed characteristic pathological changes of diabetes. TLR treatment (middle and high dose) obviously decreased serum AST, ALT levels(P<0.05)and ameliorated ultrastructure changes of liver tissues: glycogen granules decreased in cytoplasm, mitochondrial cristae became distinct and electron density decreased, appearance of smooth and rough endoplasmic reticulum became nearly normal, and the interstitial collagenofiber decreased. The SOD and CAT activities were significantly decreased and MDA levels were increased in liver tissues of diabetic rats (P<0.01). Twelve weeks of treatment with TLR obviously improved SOD activity (at all three doses, P<0.01) and decreased the levels of MDA (middle and high dose, P< 0.01). Conclusion: Our data suggest that TLR can improve hepatic injury in streptozotocin-induced diabetic rats, probably through increasing activities of antioxidant and depleting lipid peroxidation production.
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s:Objective To explore effect of the Tangshen Tongluo (TSTL) recipe on ET and ANP in patients with early diabetic nephropathy (DN). Methods 45 Cases of early DN patients were randomly divided into the TSTL recipe group (25 cases) and Lotensin group (20 cases). After the treatment of TSTL recipe and Lotensin, TCM symptom, the level of FBG, HbA1c, ET, ANP, TC, TG, HDL-C, LDL-C and urine micro albumen excretion rate (UAER) were detected. Results After the treatment of TSTL recipe and Lotensin, TCM symptom, the level of FBG, ET, ANP, TC, TG, LDL-C and UAER were found markedly lower in the TSTL recipe than that in the Lotensin group (P