[Effects of ferulic acid on inflammation and autophagy levels in glomerular mesangial cells induced by high glucose].

OBJECTIVE: To investigate the protective effects and possible mechanisms of ferulic acid on diabetic nephropathy by observing the effects of ferulic acid on the level of inflammation and autophagy in glomerular mesangial cells induced by high glucose. METHODS: SV40 MES 13 cells were cultured and randomly divided into the following groups: normal group (Control, 5.6 mmol/L glucose), mannitol group (Man, 30 mmol/L mannitol), high glucose group (HG, 30 mmol/L glucose), ferulic acid group (FA, 30 mmol/L glucose + 12.5, 25, 50, 100, 200 µmol/L ferulic acid), and the proliferation of SV40 MES 13 cells in each group was observed by MTT method. The levels of tumour necrosis factor-α (TNF-α), monocyte chemotactic protein-1 (MCP-1) and interleukin 1ß(IL-1ß)in cell supernatant were determined by enzyme-linked immunosorbent assay (ELISA). The expressions of NLRP3, IL-1ß, LC3-II/I and p62 proteins in SV40 MES 13 cells were detected by Western blot. RESULTS: ①The proliferative activity of SV40 MES 13 cells was significantly higher in the HG group compared to the control group (P＜0.01), while the proliferative activity of SV40 MES 13 cells was decreased to different degrees in the FA group compared to the HG group (P＜0.05～0.01). ②Compared to the control group, the levels of TNF-α, MCP-1 and IL-1ß were increased significantly in the cell supernatant of HG group (P＜0.01). Compared with the HG group, the levels of TNF-α, MCP-1 and IL-1ß were decreased significantly in the FA group (P＜0.01). ③Compared with the control group, LC3-II/Ⅰ protein expression was decreased in the HG group, while the levels of p62, NLRP3 and IL-1ß protein were increased significantly (P＜0.01). Compared with the HG group, the expression of LC3-II/Ⅰ protein was elevated significantly (P＜0.05) in the FA group, while the levels of p62, NLRP3 and IL-1ß protein in the FA group were decreased significantly (P＜ 0.01). CONCLUSION: FA can inhibit the abnormal proliferation of SV40 MES 13 cells induced by high glucose. FA can protect glomerular mesangial cells by inhibiting inflammation and increasing the level of autophagy.

[Effects of ketoconazole on physiological function of liver and testis in mice].

Objective: To investigate the effects of different doses of ketoconazole (KCZ) on the physiological functions of the liver and testis in Kunming mice. Methods: Forty male Kunming mice were randomly divided into four groups (n=10): normal group, KCZ low-dose group (30 mg/kg), medium-dose group (50 mg/kg), and high-dose group (70 mg/kg). The mice in the drug groups were injected subcutaneously (0.1 ml/10 g) with the corresponding dose of KCZ once a day, and the concentrations of KCZ in the KCZ low, middle, and high dose groups were 3 mg/ml, 5 mg/ml and 7 mg/ml respectively, and the normal group was injected with the same amount of normal saline for 3 weeks. The activities of aspartate transaminase (AST) and alanine aminotransferase (ALT) in serum, and γ-glutamyl transpeptidase (γ-GT), lactate dehydrogenase (LDH) and acid phosphatase (ACP) in testicular tissue were measured. HE staining was used to observe the pathological changes of the liver and testis. Results: Compared with the normal group, the activities of AST and ALT were increased significantly (P＜0.01), and the activities of γ-GT, ACP and LDH were decreased markedly in KCZ groups (P＜0.01). KCZ could affect the above indexes in a dose-dependent manner. HE staining showed that the hepatocytes were denatured, arranged loosely, and the cytoplasm was light in color. The lumen of the seminiferous tubules of the testis were enlarged, and the number of spermatogenic cells and sperm at all levels were decreased. Conclusion: KCZ could cause physiological function damage and pathological histological changes of the liver and testis, increase the levels of liver transaminase, reduce the activities of testicular specific enzymes of mice. Besides, the degree of damage was increased with the increase of dose.

Icariin ameliorates streptozocin-induced diabetic nephropathy through suppressing the TLR4/NF-κB signal pathway.

Diabetic nephropathy (DN) is one of the complex and severe complications of diabetes mellitus (DM). Icariin (ICA) is a flavonoid extracted from the leaves and stems of Herba epimedii with a wide range of pharmacological effects, such as anti-osteoporosis, anti-fibrosis, anti-aging, anti-inflammation and antioxidation. The purpose of our study was to explore the renal protective effect of ICA on DN in mice and its possible mechanisms. ICR mice were exposed to STZ-induced DN. The kidney organ coefficient of mice was computed. 24 h UP in urine was measured. Serum FBG, Cr and BUN were detected. The content of MDA and the activities of SOD, CAT and GSH-Px in renal tissues were tested. HE staining, PAS staining, PASM staining and transmission electron microscopy were used to observe renal pathological changes. Furthermore, TLR4, p-NF-κB p65, TNF-α and IL-6 of renal tissues were assayed by immunohistochemistry and western blotting. Our results indicated that ICA observably optimized the renal organ coefficient, reduced the level of 24 h UP in urine, decreased the content of Cr, BUN in serum and MDA in renal tissues, promoted the activities of SOD, CAT and GSH-Px in renal tissues, and ameliorated pathological lesions of kidneys noticeably. Besides, ICA inhibited the expressions of TLR4, p-NF-κB p65, TNF-α and IL-6 remarkably in renal tissues. ICA, which might lighten the renal inflammatory response by suppressing the TLR4/NF-κB signal pathway, played a protective role in kidneys of STZ-induced DN mice.

CPU86017, a berberine derivative, attenuates cardiac failure through normalizing calcium leakage and downregulated phospholamban and exerting antioxidant activity.

AIM: To investigate whether CPU86017, a berberine derivative, attenuates heart failure by blocking calcium influx and exerting its antioxidant activity. METHODS: Myocardial infarction was induced in male Sprague-Dawley rats for 17 d followed by isoproterenol (ISO) (5 mg/kg, sc) treatment for 5 d to reduce cardiac function. The rats were divided into 5 groups: sham operation, myocardial infarction (MI), MI plus ISO, and co-treated (in mg/kg, po) with either propranolol (PRO, 10) or CPU86017 (80). Hemodynamic measurements were conducted, and measurements of the redox system, calcium handling proteins and endothelin (ET) system in vivo were done. Furthermore, calcium flux studies and PLB immunocytochemistry were conducted in vitro. RESULTS: Compared to sham operation, HF was evident following MI and further worsened by ISO treatment. This occurred in parallel with downregulated mRNA and protein production of SERCA2a, PLB, and FKBP12.6, and was associated with upregulation of preproET-1, endothelin converting enzyme, and PKA mRNA production in the myocardium in vivo. Calcium leakage was induced by ISO treatment of isolated beating myocytes in vitro. These changes were attenuated by treatment with either PRO or CPU86017. PLB fluorescence in myocytes was downregulated by ISO treatment, and was relieved significantly by treatment with antioxidant aminoguanidine, ascorbic acid or CPU86017 in vitro. CONCLUSION: HF, calcium leakage, downregulated PLB, FKBP12.6, SERCA2a production, and upregulated PKA were caused by ISO treatment, and were abolished by CPU86017 treatment. The beneficial effects of CPU86017 are attributable to its antioxidant and calcium influx blocking effects.

Protective effect of ferulic acid on STZ-induced diabetic nephropathy in rats.

Diabetic nephropathy (DN) is a major and severe complication of diabetes mellitus. Ferulic acid (FA), a phenolic compound widespread in fruits and plants, displays a variety of pharmacological activities including regulating blood glucose and lipids, anti-oxidation, anti-inflammation and anti-fibrosis. The study was aimed to investigate the renal protective effects of FA on diabetic rats and elucidate the underlying mechanisms. FA (100 mg kg-1, i.g., once a day) was administered to DN rats for 8 weeks. The organ coefficient of kidneys was calculated. Levels of UP, BUN, Cr, FBG, TC and TG in serum were measured. Activities of SOD, CAT and GPx and the content of MDA in renal tissues were assayed. Pathological changes in renal tissues were observed by HE staining, PAS staining, PASM staining, Masson staining and transmission electron microscopy. p-NF-κB p65, TNF-α, TGF-ß1, collagen IV, nephrin and podocin protein expressed in renal tissues were determined by immunohistochemistry and western blotting. Results showed that FA significantly improved the kidney organ coefficient, decreased the UP, BUN, Cr, FBG, TC and TG levels in serum, increased SOD, CAT and GPx activities, reduced MDA content in renal tissues and alleviated pathological injury of the renal tissues. What's more, long-term treatment with FA considerably down-regulated the expressions of p-NF-κB p65, TNF-α, TGF-ß1 and collagen IV proteins, and up-regulated the expressions of nephrin and podocin proteins in renal tissues. FA could be a renoprotective agent by attenuating oxidative stress, inflammation, and fibrosis, as well as improving podocyte injury in STZ-induced DN rats.

[Selection of modeling time for type 2 diabetes mellitus mouse].

OBJECTIVE: To analyze the changes of blood biochemical index and the pathological changes of myocardium and kidney in type 2 diabetic mouse at different time points, which can provide the basis for the selection of type 2 diabetic modeling time for later research. METHODS: After 6 weeks of feeding with high-fat diet, 24 healthy male ICR mice were injected with streptozocin (STZ, 30 mg/kg) intraperitoneally for 5 days to establish diabetic models. After 9 days, a random blood glucose ≥ 11.1 mmol / L was measured as diabetic mice. 4, 6 and 8 weeks after successfully preparing the diabetic mouse, 8 diabetic mice (a group)would be sacrificed each time. Then the biochemical and pathological conditions were analyzed: ① the indexes of heart and kidney were calculated. ②the serum levels of creatine kinase (CK), lactate dehydrogenase (LDH), creatinine (Cr) and blood urine nitrogen (BUN) were determined. ③ Histopathological changes of myocardium and renal tissues were observed by hematoxylin and eosin (HE) staining. Masson staining was used to observe the fibrosis of myocardium. PAS staining was adopted to observe the pathological changes of renal tissue. In addition, 8 ICR male mice were taken as the control group. RESULTS: At the 4th, 6th and 8th week, cardiac organ coefficient, the values of LDH and CK were all increased compared with the control group. Cardiomyocyte hypertrophy and myocardial fibrosis could be observed. Renal organ coefficient, the values of Cr and BUN were increased. Glomerular hypertrophy, basement membrane thickening and atrophy could be perceived. CONCLUSION: At the 6th week, related biochemical and pathological changes in diabetic mice were comparatively obvious and breeding time was relatively short. Thus, 6 weeks after the preparation of the diabetic mice would be the optimal time for type 2 diabetes mellitus modeling, proper for inventions of drugs and other research purposes including pathology, physiology, biochemistry, etc.

Total triterpene acids, active ingredients from Fructus Corni, attenuate diabetic cardiomyopathy by normalizing ET pathway and expression of FKBP12.6 and SERCA2a in streptozotocin-rats.

Total triterpene acids (TTAs) isolated from Cornus officinalis Sieb., one of the herbs contained in Liuwei Dihuang decoction, were aimed at alleviating diabetic cardiomyopathy. We hypothesized that the benefits of TTAs may result from suppressing the endothelin-reactive oxidative species (ET-ROS) pathway in the myocardium. Diabetes was produced by a single injection of streptozotocin (STZ, 60 mg kg(-1), i.p.) in rats. Assessment of cardiac function, calcium handling proteins, endothelin-1 (ET-1) and redox system was conducted 8 weeks after STZ injection. Medication with TTAs (50 mg kg(-1), i.g.) was installed in the last 4 weeks. The compromised cardiac function was characterized by depressed contractility (LVSP and LV+dp/dt(max)) and relaxation (LVEDP and -LVdp/dt(min)) in association with hyperglycaemia (30.2 +/- 2.6 mmol L(-1)) in STZ-injected rats. Down-regulated expression of FKBP12.6 (calstabin 2), sarcoplasmic reticulum Ca(2+)-ATPase 2a (SERCA2a) and phospholamban (PLB) were also found. These changes occurred in connection with an increased ET-1, up-regulated mRNA of propreET-1 and endothelin converting enzyme (ECE), and a state of oxidant stress was found by increased malondialdehyde (MDA), decreased superoxide dismutase (SOD) and glutathione peroxidase (GSH-px) activity, and an enhanced activity and expression of inducible nitric oxide synthase (iNOS) in the diabetic myocardium. After 4 weeks of treatment with TTAs, these changes were alleviated dramatically despite a mild reduction in hyperglycaemia (26.9 +/- 3.4 mmol L(-1)). In conclusion, TTAs, as active ingredients of Liuwei Dihuang decoction, alleviated diabetic cardiomyopathy by normalizing the abnormality of FKBP12.6 and SERCA2a and ET-ROS pathway in the myocardium rather than by hypoglycaemic activity.

[Effects of ursolic acid on liver injury and its possible mechanism in diabetes mellitus mice].

OBJECTIVES: To study the effects of ursolic acid on liver injury in diabetic mice induced by high-fat diet combined with streptozotocin(STZ), and to explore its possible mechanisms. METHODS: Diabetes mellitus was induced in twenty male ICR mice by a combination of high-fat diet for 6 weeks with low-dose streptozotocin (30 mg/kg, i. p.) for 5 consecutive days. After 9 days, fasting blood glucose levels were determined. Mice with fasting blood glucose levels exceeded 11. 1 mmol/L were diagnosed as diabetic mice and selected for further experiment. These mice were randomly divided into two groups(each group of 10):diabetic group, ursolic acid group (100 mg/kg, i. g.), and another 10 mice were set as control group. After continuous administration for 8 weeks, body weight (BW) were weighed, fasting blood glucose (FBG), total cholesterol (TC), triglyceride (TG), alanine aminotransferase (ALT), aspartate transaminase (AST) in serum and superoxide dismutase (SOD), malondialdehyde (MDA) in liver were measured. HE staining was used to observe pathological changes of liver tissue. RESULTS: Compared with the control group, the level of FBG, TC, TG, ALT, AST, MDA were dramatically increased (P<0. 05, P<0. 01) and SOD was markedly decreased (P<0.01) in the diabetic group; HE staining showed that parts of liver cells swelled and had a light fatty degeneration as well as lymphocyte infiltrated around the portal area in model group. Compared with the diabetic group, the level of FBG, TC, TG, ALT, AST, MDA were significantly declined (P<0.05, P<0.01) and SOD was considerably increased (P<0.01) in the ursolic acid group; HE staining showed that the liver cells relatively arranged in order, edema was not obvious and inflammatory cells infiltrated lightly in the ursolic acid group. CONCLUSIONS: Ursolic acid has a protective effect on liver injury in diabetic mice induced by high-fat diet combined with STZ by intraperitoneal ingector, and its mechanism may be associated with lowering blood glucose, regulating the lipid metabolism, reducing oxidative stress and enhancing the ability of anti-oxidation in liver.

Ursolic acid ameliorates oxidative stress, inflammation and fibrosis in diabetic cardiomyopathy rats.

Diabetic cardiomyopathy is a major and severe cardiovascular complication of diabetes mellitus. Ursolic acid, a pentacyclic triterpene compound widespread in fruits and plants, performs a variety of pharmacological activities including lowering blood glucose, anti-oxidation, anti-inflammation and anti-fibrosis. Our present study aimed to investigate the cardioprotective effects of ursolic acid on diabetic cardiomyopathy rats and uncover its underlying mechanism. Diabetes mellitus was induced by a single injection of STZ-only (40 mg/ kg, i.v.) in male SD rats. Animals were divided into three groups (n=10): control group (normal saline, i.g.), diabetic group (normal saline, i.g.) and diabetic+ursolic acid group (35 mg/kg UA + normal saline, i.g.). Rats were administered for 8 weeks from 5th to 12th week. After the last administration, cardiac function was evaluated; HWI was calculated; FBG, CK, LDH in serum and SOD, MDA in cardiac tissue were detected. HE staining and Masson trichrome staining were employed to observe pathological alterations. Immunohistochemistry and western blotting were taken to determine the expression levels of TNF-α, MCP-1, TGF-ß1 and MMP-2 in the heart. The results dramatically showed increased levels of FBG, CK, LDH, MDA and a decreased activity of SOD in diabetic group, in which left ventricular dysfunction, cardiac myocytes hypertrophy, inflammatory cell infiltration and myocardial interstitial fibrosis had also been found. What's more, the expressions of TNF-α, MCP-1 and TGF-ß1 were significantly up-regulated and the expression of MMP-2 was markedly down-regulated in myocardium. Interestingly, treatment with ursolic acid remarkably ameliorated these changes. Collectively, our study strongly showed that ursolic acid is capable of improving the cardiac structure and function in STZ-induced diabetic cardiomyopathy rats by attenuating oxidative stress, inflammation and fibrosis.

Ursolic acid improves diabetic nephropathy via suppression of oxidative stress and inflammation in streptozotocin-induced rats.

Inflammation plays a pivotal role in the pathogenesis of diabetic nephropathy (DN). Overexpression of inflammatory chemokine and cytokines is involved in the development of DN. Ursolic acid (UA), a common pentacyclic triterpenoid compound, has been reported to have myriad benefits and medicinal properties. However, its protective effects against renal injury in streptozotocin (STZ)-induced diabetic rats have not been firmly established. In the current report, we investigated whether UA inhibits oxidative stress and inflammation in the kidneys of STZ-induced diabetic rats. Diabetes mellitus (DM) was induced by STZ (40 mg/ kg, i.v.). Animals were randomly divided into control group (normal saline, i.g.), DN group (normal saline, i.g.), DN + UA group (35 mg/kg UA + normal saline, i.g.) and DN + telmisartan group (12 mg/kg telmisartan + normal saline, i.g.). Fasting blood glucose (FBG) levels were monitored at regular intervals. The administration of compounds started at 5th week and lasted for 8 weeks. At the beginning of 13th week, rats were humanely euthanized, KW/BW, BUN, SCr, SOD and MDA were measured. Histopathological changes in renal tissue were observed after hematoxylin-eosin (HE) staining. Furthermore, the expressions of TNF-α, MCP-1 and IL-1ß in kidney were determined by immunohistochemistry and western blot. Our results showed that UA significantly lowered the levels of FBG, KW/BW, BUN, SCr and MDA in diabetic rats. Additionally, the SOD activity in UA treated group was higher than that in DN group. Furthermore, renal structural abnormalities and the elevation of TNF-α, MCP-1 and IL-1ß expression level were blocked by the administration of UA. In conclusion, our data demonstrate that UA could be well used as a protective agent to counter renal dysfunction - through antioxidant and anti-inflammatory effects.

[Effect of ursolic acid on cardiomyopathy of mice with diabetes and its mechanism].

OBJECTIVE: To study the effect of ursolic acid on cardiomyopathy in mice with diabetes induced by high-fat diet combined with low dose streptozotocin, and to explore its possible mechanism. METHODS: Thirty male ICR mice were randomly divided into control group (n=10) and moulding group (n=20), the mice in the two groups were fed with regular diet and high-fat diet respectively for 6 weeks, and then the mice in the moulding group were injected with streptozotocin (30 mg/kg) for 5 successive days to induce diabetes mellitus (DM). Fasting blood glucose (FBG) was measured after 9 days. Mice with FBG over 11.1 mmol/L were regarded as DM. Twenty DM mice were randomly divided into model group and ursolic acid group (n=10). Mice in each group were continuously administrated ursolic acid (100 mg/kg) or corresponding solvent intragastrically for 8 weeks. After that, FBG was measured, body weight (BW), heart weight and left ventricular weight were weighed in order to calculate the heart mass index (HMI) and left ventricular mass index (LVMI). Levels of creatine kinase (CK), lactate dehydrogenase (LDH) in serum and the level of superoxide dismutase (SOD), malondialdehyde (MDA) in myocardial tissue were detected. HE staining was used to observe pathological changes of myocardial tissue. Immunohistochemistry was employed to determine the expression of NOD-like receptor protein 3 (NLRP3) and interleukin 1ß (IL-1ß). RESULTS: Compared with the control group, HMI, LVMI were apparently enlarged, levels of FBG, CK, LDH in serum and MDA in myocardial tissue were extremely increased, while the activity of SOD in myocardial tissue were extraordinary decreased in diabetic group. HE staining of myocardium showed that arrangement disorder of myocardial fibers, edema and hypertrophy in myocardial cell, as well as inflammatory cell infiltration in model group. Immunohistochemistry showed that the expression of NLRP3 and IL-1ß in myocardial tissue increased obviously in model group, the above changes inursolic acid group were significantly ameliorated. CONCLUSIONS: Ursolic acid has a obvious protective effect on myocardial injury in mice with diabetes induced by high-fat diet combined with low dose streptozotocin, and its mechanism may be associated with inhibiting NLRP3 inflammasome activation, reducing IL-1ß generation and alleviating myocardial inflammatory injury.

[Effects of ferulic acid on the expressions of nephrin and podocin in podocytes of diabetic rats].

OBJECTIVE: To investigate the effects of ferulic acid (FA) on the streptozocin (STZ) -induced kidney injury in diabetic rats and its possible mechanisms. METHODS: Diabetes was induced in male SD rats by an injection of STZ (40 mg/kg,i.v.). After 72 hours, blood glucose levels were detected and blood glucose levels exceeded 16.7 mmol/L were diagnosed as diabetic model rats. Diabetic model rats were randomly divided into model group and FA group, ten animal in each group. Another 10 healthy male SD rats were treated as control group. The rats in FA group were treated with FA (100 mg/kg, i.g.,qd) from the 5th week since the diabetic rats model was successfully established and lasted for 8 weeks. The levels of blood glucose, body weight, organ coefficient of kidney, blood urea nitrogen and creatinine were tested. HE staining was employed to observe the pathological changes of the renal tissue. Immunohistochemistry was employed to determine the protein of nephrin and podocin. RESULTS: Compared to control group, the levels of blood glucose, organ coefficient of kidney, blood urea nitrogen(BUN) and serum creatinine(sCr) were increased significantly. Renal cells from model group rats showed atrophied and disordered after HE staining and interstitial proliferation were also appeared in renal tissue of the model group. Meantime, the levels of nephrin and podocin protein were obviously decreased. These changes were significantly attenuated in the model group treated with FA. CONCLUSIONS: FA can evidently ameliorate renal damage in rats with diabetic nephropathy induced by STZ, which might be related to increase the level of nephrin and podocin protein.

[Study on the protective effect of ursolic acid on alloxan-induced diabetic renal injury and its underlying mechanisms].

OBJECTIVE: To investigate the effect of ursolic acid (UA) on the alloxan-induced kidney injury in diabetic mice and explored its possible mechanisms. METHODS: Diabetes mellitus was induced in male Kunming mice by an injection of alloxan (70 mg/kg, i.v.). After 72 hours, blood glucose levels were detected and mice with blood glucose levels over 13.9 mmol/L were considered as diabetic and selected for further experiment. Thirty mice were randomly divided into three groups: control, diabetic and diabetic + UA(35 mg/kg/d, i.g. continuously for 8 weeks). Blood glucose concentration, organ coefficient of kidney, blood urea nitrogen (BUN), creatinine (Cr) as well as renal tissue levels of superoxide dismutase (SOD), methane dicarboxylic aldehyde (MDA), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were determined. Pathology of the renal tissue was measured by hematoxylin-eosin staining. RESULTS: Compared to the control group, blood glucose, organ coefficient of kidney, BUN and Cr increased significantly. In addition, SOD activities was reduced markedly and levels of MDA and inflammatory factors (TNF-α, IL-6) increased significantly. Renal cells from model group rats showed atrophy and disordered after HE staining and infiltration of inflammatory cells also appeared in renal tissue of the model group. These changes were significantly attenuated in the diabetic group treated with UA. CONCLUSION: UA can significantly relieve renal damage in mice with diabetic nephropathy induced by alloxan, which might be related to decreased blood glucose level, antioxidation effect and inhibiting the production of inflammatory factors such as TNF-α and IL-6.

Total triterpene acids, isolated from Corni Fructus, ameliorate progression of renal damage in streptozotocin-induced diabetic rats.

OBJECTIVE: To investigate whether total triterpene acids (TTAs), isolated from Cornus Fructus, attenuates renal function by reducing oxidative stress and down-regulating the expression of transforming growth factor ß1 (TGF-ß1). 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-ß1. 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-ß1. CONCLUSION: TTAs improved renal function via reducing oxidative stress and down-regulation the expression of TGF-ß1 in diabetic rats.

[Protective effect of total flavonoids of epimedium on the kidney in experimental diabetic rats].

OBJECTIVE: To investigate the influence of total flavonoids of epimedium (TFE) on the streptozocin (STZ)-induced kidney injury in diabetic rats and discuss the possible mechanism. METHODS: Diabetes was produced by a single injection of streptozocin (40 mg/kg, iv) in male SD rats. The rats were randomly divided into three groups (n = 10): control group, model group and TFE group (100 mg/kg, ig). Animals were sacrificed 12 weeks later. The level of blood glucose, blood urea nitrogen (BUN) and creatinine (Cr) as well as the renal index were determined. Detect the specific biochemical of renal tissue: superoxide dismutase (SOD), malondialdehyde (MDA). Use masson staining to observe the morphology of the renal tissue. Immunohistochemistry was employed to determine the protein levels of transforming growth factor-beta1 (TGF-beta1). RESULTS: Compared to control group, the enhancement of blood glucose, renal index, BUN and Cr was found in model group, which was significantly attenuated by treatment with TFE. Meanwhile, elevated MDA level in renal tissue as well as decreased SOD activities in renal tissue were significantly remitted by TFE. Furthermore, TFE decreased the expression of TGF-beta1. CONCLUSION: TFE can evidently relieve renal damage in rats with diabetic nephropathy induced by STZ, which might be related to antioxidation and modulating the expression of TGF-beta1 protein.

[Study on the effect of ursolic acid (UA) on the myocardial fibrosis of experimental diabetic mice].

OBJECTIVE: To investigate the effect of ursolic acid (UA) on the alloxan-induced myocardial fibrosis in mice and discuss the possible mechanism. METHODS: Diabetes was produced by a single injection of alloxan (70 mg/kg, i.v.) in mice. The mice were randomly divided into four groups: normal control group, model group, ursolic acid group (UA, 35 mg/kg, p.o.) and benazepril group (5 mg/kg, p. o.), and continuous administrated for 8 weeks. The blood glucose was measured 24 hours after the last administration. Detected the specific biochemical of myocardial tissue: superoxide dismutase (SOD), malondialdehyde (MDA) and hydroxyproline(HYP). Using masson staining to observe the morphology of the myocardial tissue. Immunohistochemistry was employed to determine the protein levels of TGF-beta1. RESULTS: Compared to normal group, the blood glucose, heart index, myocardial tissue MDA, HYP level were increased, and SOD activities were decreased in the diabetic mice, Masson stain showed that myocardial cells disarranged, myocardial collagen fibrosis hyperplasia. Meanwhile, the protein expression of TGF-beta1 was increased in model group. The UA group improved all the above significantly. CONCLUSION: UA improves the myocardial collagen fibrosis in diabetic mice induced by alloxan, its mechanism may be related to inhibiting the expression of TGF-beta1 and antioxidation.

[Protective effect of total flavonoids of herba epimedii on testis degeneration in diabetic mice].

OBJECTIVE: To observe the protective effects and investigate the possible mechanism of total flavonoids of herba epimedii (TFE) on diabetic testopathy in mice. METHODS: Diabetic animal model was produced by a single injection of alloxan ( 70 mg/kg, i.v.) in mice. The mice were randomly divided into 3 groups (n = 10): control group, model group and TFE group (100 mg/kg, p.o.), administrated for 8 weeks continuously. The level of serum testosterone and blood glucose were measured after 24 hours in the last administration. Detect the specific biochemical indicators of testis: superoxide dismutase (SOD), malondialdehyde (MDA). Meanwhile, the morphology of testis was observed under light microscope by HE and MASSION dyeing. Immunohistochemistry was employed to detect the level of matrix metalloprotein (MMP)9. RESULTS: Compared with control group, glucose and the content of MDA in testicular tissues increased while the levels of serum testosterone and SOD decreased remarkably in model group. Detection of pathology showed that the diameters of seminiferous tubules, various grades of spermatogenic cell decreased and collagen fibrosis hyperplasia in testicular tissues, the expression of (MMP9) were decreased in model group. These alterations were significantly improved in TFE group (P < 0.01). CONCLUSION: TFE ameliorated the alterations of testis inalloxan-induced mice through promoting the testosterone release, anti-oxidation and improving the expression of MMP9.

Synthesis and biological activity of nitric oxide-releasing derivatives of ferulic acid as potential agents for the treatment of chronic kidney diseases.

In order to search for novel potential agents for the treatment of chronic kidney diseases (CKD), nitric oxide (NO)-releasing derivatives (5a-c) of ferulic acid were synthesized and characterized by MS, 1H NMR, and elementary analysis. They showed different NO-releasing rate in the absence or presence of L-cysteine in vitro. In the adenine induced CKD rats, these compounds revealed reno-protective effect via lowering blood urea nitrogen (BUN), creatinine (Cr) in serum and malondialdehyde (MDA) in kidney, increasing NO and superoxide dismutase (SOD) level in kidney. Among them, 3-methoxy-4-(nitrooxy)ethoxy cinnamic acid (5a) was confirmed to have a higher NO-releasing rate in vitro and better effect in ameliorating adenine-induced kidney damage in rats.

[Amelioration of icariin for the epididymis impairment induced by streptozocin (STZ) in rats].

OBJECTIVE: To investigate the effects of icariin on the streptozocin (STZ)-induced epididymis impair in rats. METHODS: The epididymis impair was induced by injection of streptozocin at dosage of 60 mg/kg ip in SD rats. Animals were randomly divided into six groups (n = 14): normal control, model group, three icariin treated group with different dosages (P.O, 20 mg/kg, 40 mg/kg, 80 mg/kg especially) and rosiglitazone group (P.O, 3 mg/kg), 12 weeks later, animals were sacrificed. The level of serum glucose, the activity of lactate dehydrogenase (LDH), acid phosphatase (ACP), gamma-glutamyltranspeptidase (gamma-GT), alpha-glucosidase activity as well as sialic acid (SA), fructose level in the epididymis were determined. The pathological examination was performed under microscope after the epididymis was fixed by 4% poly-formalin and stained by HE. RESULTS: Compared with the normal control, the activity of LDH, ACP, gamma-GT and alpha-glucosidase in the epididymis revealed a decline, with lower level of SA and fructose. Histological examination showed that mature spermatocytes in the epididymis markedly decreased. These alterations were ameliorated in the groups with the treatment of icariin at 40 mg/kg and 80 mg/kg, but not at 20 mg/kg. CONCLUSION: Icariin ameliorated the signs of epididymis in diabetic rats induced by streptozocin, this effect might carry out by promoting the elevation of special enzyme and energy metabolism in the epididymis.

[Protective effect of terpenes from fructus corni on the cardiomyopathy in alloxan-induced diabetic mice].

OBJECTIVE: To investigate the protective effects of terpenes from fructus corni (TFC) on diabetic cardiomyopathy (DCM). METHODS: Diabetes was produced by a single injection of alloxan (220 mg/kg, i.p.) in mice. The fasting blood glucose of mice were tested 15 days later and that greater than 13.9 mmol/L were regarded as the diabetic mice which were divided randomly into the model and TFC groups. TFC dissolved by physiological saline (P.O, 80 mg/kg) was administrated to the TFC group for successive 8 weeks since the 15th day. RESULTS: Compared to the control group, the weight index increased significantly. The level of superoxide dismutase (SOD) was markedly decreased and malondialdehyde(MDA), the inflammatory factors (TNF-alpha, IL-6) were obviously increased in myocardium. The histopathological examination suggested that myocardial cells disarranged, swelling and the intercellular space increased in model group. It also showed the infiltration of inflammatory cells and fibroblasts in TFC group. The above change was improved significantly. CONCLUSION: TFC ameliorated the alterations of cardiomyopathy in diabetic mice induced by alloxan. the mechanism might be related to decrease blood glucose, antioxidative stress and inflammatory factors.