Ferulic acid in the treatment of post-diabetes testicular damage: relevance to the down regulation of apoptosis correlates with antioxidant status via modulation of TGF-ß1, IL-1ß and Akt signalling.

The aim of this study was to investigate the protective effect of ferulic acid at different doses (50 mg kg(-1) alternative day and 50 mg kg(-1) daily) on the streptozotocin (STZ)-induced post-diabetes rat testicular damage. Diabetes was induced by a single intraperitoneal injection of STZ (50 mg/kg). Rats treated with ferulic acid were given once a day orally for 10 weeks, starting 3 days after STZ injection. Testis tissue and blood samples were collected for investigating biochemical analysis, antioxidant status, sperm parameters, and histopathological, immunohistochemical and apoptotic studies. Treatment with ferulic acid to diabetic rats significantly improved the body weight, testis weight, serum insulin level, serum testosterone level and sperm parameters (viability, motility and count). Histopathological study also revealed that ferulic acid-treated diabetic rats showed an improved histological appearance. Our data indicated that significant reduction in the activity of apoptosis by using terminal deoxyuridine triphosphate nick end-labelling and reduced expression of transforming growth factor-ß1 and interleukin-1ß in the testis tissue of ferulic acid-treated diabetic rats. Conversely, it was also revealed that ferulic acid-treated diabetic rats markedly enhanced the serine/threonine protein kinase protein expression in the testis tissue. Our result suggests that ferulic acid inhibits testicular damage in diabetic rats by declining oxidative stress.

Treatment with ferulic acid to rats with streptozotocin-induced diabetes: effects on oxidative stress, pro-inflammatory cytokines, and apoptosis in the pancreatic ß cell.

In the present study, we aimed to investigate the protective effect of ferulic acid at different doses (50 mg/kg alternative day and 50 mg/kg daily) on diabetic rats and to explore the interrelationship between oxidative stress and cytokines correlates with apoptotic events in pancreatic tissue. Male Wistar rats were rendered diabetic by a single intraperitoneal injection of streptozotocin (60 mg/kg body weight). Ferulic acid was administered orally for 8 weeks. At the end of the study, all animals were sacrificed. Blood samples were collected for the biochemical estimations and pancreas was isolated for antioxidant status, histopathological, immunohistochemical, and apoptotic studies. Treatment with ferulic acid to diabetic rats significantly improved blood glucose, serum total cholesterol, triglycerides, creatinine, urea, and albumin levels toward normal. Furthermore, decrement of the elevated lipid peroxidation levels and increment of the reduced superoxide dismutase, catalase, and reduced glutathione enzyme activities in pancreatic tissues were observed in ferulic acid-treated groups. Ferulic acid-treated rats in the diabetic group showed an improved histological appearance. Our data also revealed a significant reduction in the activity of apoptosis using terminal dUTP nick end-labeling and reduced expression of TGF-ß1 and IL-1ß in the pancreatic ß-cell of ferulic acid-treated rats. Treatment with ferulic acid daily doses produced a significant result compared to alternative dose. Collectively our results suggested that ferulic acid acts as a protective agent in diabetic rats by altering oxidative stress, expression of pro-inflammatory cytokines and apoptosis.

Combination therapy of dexamethasone with epigallocatechin enhances tibiotarsal bone articulation and modulates oxidative status correlates with cartilage cytokines expression in the early phase of experimental arthritis.

The inclusion of antioxidant for the treatment of arthritis, especially under the therapy with immunosuppressant, is motivated because antioxidant plays an essential role in disease progression and moreover, immunosuppressive treatment suffers redox homeostasis balance of the organism. The aim of the present study was to evaluate the enhancement of anti-arthritic effect of dexamethasone in combination with epigallocatechin on the progression of adjuvant-induced arthritis in rats. Adjuvant arthritic rats were treated with dexamethasone (0.2mg/kg), epigallocatechin (100mg/kg) and combination of dexamethasone (0.1mg/kg) with epigallocatechin (100mg/kg) daily for a period of 28 days. Paw swelling changes, estimation of serum albumin level, alteration of bone mineral density, histopathological, and radiographical analysis were assessed to evaluate the anti-arthritic effect. Lipid peroxidation and antioxidant enzyme activities in joint tissue homogenate were performed along with the expression of different pro-inflammatory cartilage cytokines like TNF-α and IL-6. Dexamethasone and epigallocatechin combination potentiated both the antiarthritic (decrease of hind paw volume) and the antioxidant effect (lipid peroxidation, superoxide dismutase, glutathione reductase and catalase). In combination with dexamethasone, epigallocatechin markedly potentiated the beneficial effect of dexamethasone which resulted in more significant increment of serum albumin and bone mineral density. Improvement of anti-arthritic effect of combination therapy was supported by histopathological, radiographical alterations, and attenuation of over-expression of cartilage cytokines. Epigallocatechin act as potent antioxidant and combined administration of dexamethasone with epigallocatechin increased the anti-arthritic efficacy of basal dexamethasone therapy and suppressed the development phase of arthritic progression in rats.

A novel combination of methotrexate and epigallocatechin attenuates the overexpression of pro-inflammatory cartilage cytokines and modulates antioxidant status in adjuvant arthritic rats.

The present study was designed to evaluate the combinatory effect of methotrexate (MTX) and epigallocatechin (EGCG) on the progression of adjuvant-induced arthritis in rats. Adjuvant arthritis (AA) was induced by a single intradermal injection of Freund's complete adjuvant. AA rats were treated with methotrexate (0.3 mg/kg) thrice a week, EGCG (100 mg/kg) daily, and combination of MTX and EGCG thrice a week for a period of 28 days. Paw swelling changes and histopathological and radiographic analysis was assessed to evaluate the antiarthritic effect. Lipid peroxidation and antioxidant enzyme activities in joint tissue homogenate were performed to observe the modulation of antioxidant status along the expression of different pro-inflammatory cartilage cytokines like TNF-α and IL-6. MTX and EGCG combination potentiated both the antiarthritic (decrease of hind paw volume) and the antioxidant effect (SOD, GSH, and catalase) as well as suppression of lipid peroxidation. Combination therapy of MTX and EGCG significantly inhibited the development phase of arthritis, which is supported by histopathological, radiographical, and attenuation of overexpression of cartilage cytokines. EGCG act as potent antioxidant and immunomodulator, suggesting that combined administration of MTX along with EGCG suppressed the development phase of arthritic progression in rats.

Terpenoids of methanol extract of Clerodendrum infortunatum exhibit anticancer activity against Ehrlich's ascites carcinoma (EAC) in mice.

CONTEXT: Clerodendrum infortunatum Linn. is a widely used plant in the Indian indigenous system of medicine for the treatment of tumors. OBJECTIVE: The present study evaluated the anticancer activity of methanol extract of C. infortunatum (MECI) against Ehrlich's ascites carcinoma (EAC) bearing Swiss albino mice and isolation of bioactive terpenoids from it. METHODS: HPLC analysis of the methanol extract showed the presence of three major components. Out of those, two compounds were isolated and characterized as oleanolic acid and clerodinin A. The anticancer activity of MECI was assessed by measuring the tumor growth response, percentage increase of life span, study of hematological parameters, lipid peroxidation, antioxidant enzyme activity like glutathion and CAT. In vitro cytotoxicity assay was also performed using EAC cell lines. RESULT AND CONCLUSION: Treatment with MECI causes significant decrease in the tumor cell volume and increase in the life span. The median survival time (MST) of EAC control group was found as 19.42 ± 0.91 d, whereas the MST was increased to 23.44 ± 2.69 d and 27.57 ± 2.57 d for the groups treated with MECI at 100 and 200 mg/kg, respectively. All the hematological parameters, malonaldehyde content and antioxidant enzymes' activity were restored towards the normal level. IC(50) value of MECI was found as 498.33 µg/mL in cytotoxicity study. The experimental results suggested that MECI has significant anticancer activity, which can be attributed to the presence of oleanolic acid and clerodinin A.

Resveratrol regulates antioxidant status, inhibits cytokine expression and restricts apoptosis in carbon tetrachloride induced rat hepatic injury.

Recent studies indicate the chemopreventive role of resveratrol in many animal models like ischemia, rheumatoid arthritis, human cancer, and diabetes. The present study was designed to investigate the chemopreventive potential of resveratrol in rat hepatic injury model by carbon tetrachloride. Male Wistar rats were treated with carbon tetrachloride (0.4 g/kg body weight) intraperitoneally daily for 8 weeks. Resveratrol (100 mg/kg, 200 mg/kg body weight) was given orally from first day until the last day of experiment. The investigation assesses the effect of resveratrol on morphological, oxidative status, histopathological, immunohistochemical, and apoptotic analysis in carbon tetrachloride-challenged liver tissue. The study indicated that the inflammatory cytokines TNF-α and IL-6 were profoundly expressed in experimental rats, whereas resveratrol decreases the immunopositivity of TNF-α and IL-6 and restored the altered architectural structure of challenged hepatic tissue. Resveratrol also protects liver cells by suppressing oxidative stress and apoptosis.

Isolation and in vivo hepatoprotective activity of Melothria heterophylla (Lour.) Cogn. against chemically induced liver injuries in rats.

OBJECTIVE: To investigate hepatoprotective activity of ethanol extract of Melothria heterophylla Lour Cogn. (EEMH) against CCl(4)-induced hepatic damage in rats. METHODS: ß-sitosterol was isolated by column chromatography and characterized spectroscopically. Two different doses (200 and 400 mg/kg bw) of EEMH were administered orally in alternate days. The hepatoprotective activity was studied in liver by measuring biochemical parameters such as serum aspartate amino transferase (AST), alanine amino transferase (ALT), alkaline phosphatase (ALP), total protein and total bilirubin. Lipid peroxidation product and different antioxidant enzyme activities were assessed in liver homogenate. RESULTS: EEMH reduced all biochemical parameters and lipid peroxidation, as well as it increased the antioxidant enzyme activities in comparison with silymarin. The protective effect of the extract on CCl(4) induced damage was confirmed by histopathological examination of the liver. CONCLUSIONS: This result strongly supports the protective effect of EEMH against acute liver injury, and may be attributed to its antioxidative activity.

Flavonoids of Enhydra Fluctuans exhibits analgesic and anti-inflammatory activity in different animal models.

Enhydra fluctuans (Compositae), an edible semi aquatic herbaceous vegetable plant, widely used in traditional system of Indian medicine. Total flavonoids of E. fluctuans (TFEF) were screened for analgesic and anti-inflammatory activity. Analgesic activity was studied in acetic acid induced writhing response and by hot plate method in Swiss albino mice. Anti-inflammatory activity was estimated by carrageenan and histamine induced acute inflammation and Freund's complete adjuvant (FCA) induced chronic inflammation in rats. Two flavonoids, baicalein 7-O-glucoside and baicalein 7-O-diglucoside, were isolated from the ethyl acetate fraction. Oral administration of TFEF at the doses of 200 and 400 mg/kg provide 27.05 and 55.49% protection respectively in acetic acid induced writhing method. It also increased the pain threshold in mice evidenced by hot plate method. TFEF showed more potent anti-inflammatory activity. The results of this study may be attributed to high free radical scavenging and antioxidant potential of the flavonoids present in ethyl acetate fraction of Enhydra fluctuans.

Methoxy VO-salen stimulates pancreatic ß cell survival by upregulation of eNOS and downregulation of apoptosis in STZ-induced diabetic rats.

The present study was designed to investigate the effect of MetVO-salen in ameliorating diabetes and oxidative stress in the pancreas of diabetic rats. Streptozotocin (STZ)-induced diabetic rats were treated with MetVO-salen complex intraperitonially (0.3 and 0.6 mg/kg) thrice a week and continued for 8 weeks. Total cholesterol, high-density lipoprotein (HDL) cholesterol, triglycerides in serum, and blood glucose were estimated. Furthermore, oxidative stress in rats was also investigated in terms of superoxide dismutase (SOD), catalase, lipid peroxidation, and glutathione (GSH). In addition, the anti-diabetic activity of MetVO-salen was also investigated by assessing histopathological, immunohistochemical in terms of endothelial nitric oxide synthase expression, and apoptotic events in pancreas. Treatment with MetVO-salen complex reduced the blood glucose level and significantly altered the serum biochemical parameters of diabetic rats. Treatment with above complex decreased the lipid peroxidation and the antioxidant enzymes such as SOD, CAT, and GSH to near-control levels. Histopathological, immunohistochemical, and apoptotic studies also revealed that MetVO-salen-induced amelioration of the diabetic state appears to be significant to the preservation of a functional portion of the pancreatic ß cells which initially prevent STZ toxicity. This study provides new direction for the management of diabetes but needs further clinical evaluation.

Downregulation of apoptosis and modulation of TGF-ß1 by sodium selenate prevents streptozotocin-induced diabetic rat renal impairment.

To investigate whether sodium selenate treatment would impact on the onset of diabetic nephropathy, we examined blood glucose, serum biochemical components, and interrelationship between oxidative stress, TGF-ß1, and apoptosis in streptozotocin (STZ) induced diabetic rats. Sixty male Wistar rats were divided into six groups. Group I (n = 10), normal control; Group II (n = 10), diabetic control; Group III (n = 10), sodium selenate (16 µmoles/kg) + diabetic; Group IV (n = 10), sodium selenate (32 µmoles/kg) + diabetic; Group V (n = 10), sodium selenate (16 µmoles/kg) control; and Group VI (n = 10), sodium selenate (32 µmoles/kg) control. Sodium selenate was administered via orogastric route for 10 weeks. In the diabetic group, diabetes was induced by single intraperitoneal injection of STZ (50 mg/kg). The levels of blood glucose were estimated and total cholesterol, high-density lipoprotein (HDL) cholesterol, triglycerides, creatinine, urea, and albumin were detected in serum. Antioxidant status was examined by measuring the superoxide dismutase (SOD), catalase, glutathione, and lipid peroxidation in kidney tissues. Histopathological studies were performed in the kidney tissue sections. The expression of TGF-ß1 was estimated by the immunohistochemical analysis in kidneys. Apoptotic study in kidney was performed using the TdT-mediated dUTP nick end labeling technique. It was observed that blood glucose, serum, total cholesterol, HDL cholesterol, triglycerides, creatinine, urea, and albumin were significantly higher in diabetic control groups. Diabetic + sodium selenate (16 and 32 µmoles/kg) significantly reduced blood glucose, serum, total cholesterol, HDL cholesterol, triglycerides, creatinine, urea, and albumin levels. Selenium-treated groups significantly increased antioxidant enzyme activities (SOD, catalase, and glutathione) in kidneys of diabetic rats. All enzyme activities of selenium control groups did not differ compared with the normal control. Sodium selenate reduces significantly lipid peroxidation in diabetic rats. Cellular architecture of the diabetic rats was altered whereas sodium selenate administration rectifies the degenerative changes of the kidney. Profound immunopositivity of TGF-ß1 was observed in the glomerular and tubulointerstitial cells of diabetic rat kidney. Immunopositivity of TGF-ß1 was significantly reduced in both low and high dose of sodium-selenate-treated rats (P < 0.05, P < 0.01). High numbers of apoptotic cells were observed in diabetic rats whereas sodium selenate in both doses significantly reduces the incidence of apoptosis (P < 0.05, P < 0.01). We conclude herein that sodium selenate has the potential to play a significant role in limiting the renal impairment by altering the apoptosis and TGF-ß1 in experimental diabetic rats.

Flavonoids of Enhydra fluctuans exhibit anticancer activity against Ehrlich's ascites carcinoma in mice.

Flavonoids obtained from Enhydra fluctuans (FEF) were screened for anticancer activity against Ehrlich's ascites carcinoma (EAC) bearing Swiss albino mice. The anticancer activity was assessed by measuring the tumor growth response, percentage increase of life span, hematological parameters, lipid peroxidation, and antioxidant enzyme activity, like GSH and CAT. Two flavonoids, baicalein 7-O-glucoside and baicalein 7-O-diglucoside, were isolated from the ethyl acetate fraction. Treatment with FEF caused a significant decrease in the tumor cell volume and increase of life span. All the hematological parameters, malonaldehyde content and antioxidant enzyme activity were restored towards the normal level. FEF was found to be cytotoxic in the in-vitro model.

Antioxidant Potential of Crude Extract and Different Fractions of Enhydra fluctuans Lour.

The antioxidant potential of crude methanol extract (CE) as well as chloroform (CF), ethyl acetate (EF) and n-butanol (NF) soluble fractions of Enhydra fluctuans Lour, which is widely used in indigenous system of medicine for different purposes, were studied. The antioxidant potential of extract/different fractions were evaluated using different in vitro antioxidant models. In addition, total amount of polyphenolics compounds, DPPH (1,1-diphenyl-2-picryl hydrazyl) radical, nitric oxide, superoxide anion, hydroxyl radical scavenging activities and reductive power of crude extracted its different fractions were determined. It was found that ethyl acetate fraction have maximum amount of polyphenolics compounds (179.7 ± 18.23 µg / mg pyrocatechol equivalent). This fraction was found more effective than crude extract and other fractions in all the above mentioned assays.

Analgesic and anticonvulsant effects of saponin isolated from the leaves of Clerodendrum infortunatum Linn. in mice.

Saponin (SN1) isolated from C. infortunatum leaves in doses of 30, 50, 75 and 100 mg/kg, ip provided 36.28, 60.47, 90.71, 100% protection respectively from writhing induced by 1.2% v/v acetic acid. In hot plate method, SN1 not only produced analgesia in mice but also potentiated the analgesic action of pentazocine and aspirin. The anticonvulsant activity was tested by leptazol-induced seizures. SN1 decreased the duration of seizures and gave protection in a dose dependent manner against leptazol-induced convulsions. The results suggest that saponin has significant analgesic and anticonvulsant effects.

Kinetic measurements of the hydrolytic degradation of cefixime: effect of Captisol complexation and water-soluble polymers.

We have taken kinetic measurements of the hydrolytic degradation of cefixime, and have studied the effect of Captisol complexation and water-soluble polymers on that degradation. The phase solubility of cefixime in Captisol was determined. Kinetic measurements were carried out as a function of pH and temperature. High-performance liquid chromatography (HPLC) was performed to assay all the samples of phase-solubility analysis and kinetic measurements. Chromatographic separation of the degradation products was also performed by HPLC. FT-IR spectroscopy was used to investigate the presence of any interaction between cefixime and Captisol and soluble polymer. The phase-solubility study showed A(L)-type behaviour. The pH-rate profile of cefixime exhibited a U-shaped profile whilst the degradation of cefixime alone was markedly accelerated with elevated temperature. A strong stabilizing influence of the cefixime-Captisol complexation and hypromellose was observed against aqueous mediated degradation, as compared with povidone and macrogol. The unfavourable effect of povidone and macrogol may have been due to the steric hindrance, which prevented the guest molecule from entering the cyclodextrin cavity, whereas hypromellose did not produce any steric hindrance.

Crystallization of a non-steroidal anti-inflammatory drug from ethanol-water solution in presence of polymers: physicochemical characterization and release behaviour from suppositories.

Altered crystallization condition has been designed and adopted to a model non-steroidal anti-inflammatory drug, while crystallizing from ethanol-water solution in absence and presence of polymers such as Eudragit RS and ethylcellulose. To minimize the gastro-intestinal side effects nimesulide was considered as a model drug candidate for the development of suppository formulation. Physicochemical characteristics of the crystals were evaluated by Scanning Electron Microscopy (SEM). X-ray diffraction (XRD) and Fourier Transformed Infrared Spectroscopy (FT-IR). Smoothness and sharpness of the crystal have been decreased with increased concentration of a polymer. A little change in crystal habit and geometry has also been observed. Crystals are discrete in nature and more than 90% were in the range of 20-90 micron. The X-ray diffractions of nimesulide crystallized in absence of polymer and physical mixture of drug-polymer revealed fewer high intensity reflections when compared with the drug crystallized in presence of Eudragit RS, which testified a slight decreased ordering of crystal lattice in the latter. In presence of ethylcellulose, slightly increased ordering of crystal lattice was observed. No strong interactions were noticed as revealed by FT-IR spectroscopy. Drug dissolution rate from suppository formulations containing nimesulide crystallized in presence of polymer was found to delay as compared with the suppository prepared by nimesulide crystallized in absence of polymer.