HD-03/ES: A Herbal Medicine Inhibits Hepatitis B Surface Antigen Secretion in Transfected Human Hepatocarcinoma PLC/PRF/5 Cells.

HD-03/ES is a herbal formulation used for the treatment of hepatitis B. However, the molecular mechanism involved in the antihepatitis B (HBV) activity of this drug has not been studied using in vitro models. The effect of HD-03/ES on hepatitis B surface antigen (HBsAg) secretion and its gene expression was studied in transfected human hepatocarcinoma PLC/PRF/5 cells. The anti-HBV activity was tested based on the inhibition of HBsAg secretion into the culture media, as detected by HBsAg-specific antibody-mediated enzyme assay (ELISA) at concentrations ranging from 125 to 1000 µ g/mL. The effect of HD-03/ES on HBsAg gene expression was analyzed using semiquantitative multiplex RT-PCR by employing specific primers. The results showed that HD-03/ES suppressed HBsAg production with an IC50 of 380 µ g/mL in PLC/PRF/5 cells for a period of 24 h. HD-03/ES downregulated HBsAg gene expression in PLC/PRF/5 cells. In conclusion, HD-03/ES exhibits strong anti-HBV properties by inhibiting the secretion of hepatitis B surface antigen in PLC/PRF/5 cells, and this action is targeted at the transcription level. Thus, HD-03/ES could be beneficial in the treatment of acute and chronic hepatitis B infections.

Pharmacodynamics & toxicological profile of PartySmart, a herbal preparation for alcohol hangover in Wistar rats.

BACKGROUND & OBJECTIVE: PartySmart is a herbal preparation intended for the management of alcohol hangover and other related toxic effects in clinical situation. The present study was designed to investigate the pharmacodynamics and oral toxicity of PartySmart, a herbal formulation in rats. METHODS: Effect of PartySmart on blood acetaldehyde and alcohol levels was evaluated at doses of 125, 250 and 500 mg/kg b.wt. in rats. Acute toxicity study was conducted with PartySmart at a limit test dose of 2000 mg/kg b.wt., p.o. In repeated dose 90 day study, PartySmart was administered at doses of 500 and 1000 mg/kg b.wt. once-a-day, orally throughout the study period. RESULTS: PartySmart dose-dependently decreased blood ethanol and acetaldehyde levels as compared to control. PartySmart at a dose of 500 mg/kg b.wt. significantly reduced the area under curve (AUC) of ethanol and acetaldehyde levels. It increased the hepatic alcohol dehydrogenase (ADH) at 500 mg/kg b.wt. and aldehyde dehydrogenase (ALDH) activities at doses of 250 and 500 mg/kg b.w. significantly. Acute toxicity study showed no clinical signs and pre-terminal deaths. The LD(50) of PartySmart was found to be greater than 2000 mg/kg b.wt. No significant differences in PartySmart-treated groups were observed on body weight, food intake, haematological and clinical chemistry, and organ weight ratios as compared to control group in the repeated dose study. Histopathological examination of all target organs showed no evidence of lesions attributing to drug toxicity. INTERPRETATION & CONCLUSION: PartySmart enhanced acetaldehyde metabolism by increasing ADH and ALDH activity without any side effects. These findings indicate that PartySmart may exert beneficial role in the management of alcohol hangover without any toxicity.

Ameliorative effect of Partysmart in rat model of alcoholic liver disease.

Present study was designed to investigate the effect of polyherbal formulation PartySmart in experimental model of alcoholic liver disease in male Wistar strain rats. Alcohol plus fish oil were administered to animals for 8 weeks to induce liver injury. PartySmart was administered at doses of 250 and 500 mg/kg body weight. After 8 weeks, parameters such as liver weight, liver function serum markers alanine transaminase (ALT), aspartate transaminase (AST) and alkaline phosphatase (ALP) and lipid peroxidation were studied. Livers from all the groups were subjected for histological evaluation. Treatment with PartySmart at the dose of 500 mg/kg body weight showed significant reduction in the levels of serum ALT, AST and ALP with a decrease in liver weight as compared to ethanol-fed rats. A significant decrease was also observed in malondialdehyde levels following treatment with PartySmart at 500 mg/kg body weight. Histological profile of liver tissue in PartySmart-treated animals showed lesser vacuolar degeneration and intactness of hepatic architecture along with improved glycogen deposition as demonstrated by PAS staining. PartySmart ameliorated alcohol-induced liver injury by preventing cell membrane disturbances, reduction of oxidative stress by free radical scavenging and antioxidant activity and normalization of altered intracellular redox status. Thus, PartySmart can be beneficial in the treatment of alcohol-induced liver damage.

NCB-02 (standardized Curcumin preparation) protects dinitrochlorobenzene- induced colitis through down-regulation of NFkappa-B and iNOS.

AIM: To evaluate the efficacy and mechanism of action of NCB 02, a standardized Curcumin preparation, against 2, 4 dinitrochlorobenzene (DNCB) induced ulcerative colitis in rats. METHODS: Ulcerative colitis was induced in male rats by sensitizing with topical application of DNCB in acetone for 14 d and intra-colonol challenge with DNCB on day 15. A separate group of animals with vehicle treatment in similar fashion served as control group. Colitis rats were divided into different groups and treated with NCB-02 at doses of 25, 50 and 100 mg/kg b.wt p.o. for 10 d. Sulfasalazine at a dose of 100 mg/kg b.wt for 10 d served as a reference group. On day 10 after respective assigned treatment, all the animals were euthanized and the length of the colon, weight of entire colon and distal 8 cm of the colon were recorded. The distal part of the colon was immediately observed under a stereomicroscope and the degree of damage was scored. Further distal 8 cm of the colon was subject to the determination of colonic myeloperoxidase (MPO), lipid peroxidation (LPO) and alkaline phosphatase (ALP) activities. A small piece of the sample from distal colon of each animal was fixed in 10% neutral buffered formalin and embedded in paraffin wax and sectioned for immunohistochemical examination of NFkappa-B and iNOS expression. RESULTS: NCB-02 showed a dose dependent protection against DNCB-induced alteration in colon length and weight. NCB-02 treatment also showed a dose dependent protection against the elevated levels of MPO, LPO and ALP, induced by DNCB. NCB-02 demonstrated a significant effect at a dose of 100 mg/kg b.wt., which was almost equipotent to 100 mg/kg b.wt. of sulfasalazine. Treatment with sulfasalazine and curcumin at a dose of 100 mg/kg b.wt. inhibited the DNCB-induced overexpression of NFkappa-B and iNOS in the colon. CONCLUSION: Curcumin treatment ameliorates colonic damage in DNCB induced colitic rats, an effect associated with an improvement in intestinal oxidative stress and downregulation of colonic NFkappa-B and iNOS expression.

Induction and evaluation of atherosclerosis in New Zealand white rabbits.

Atherosclerosis was experimentally induced in New Zealand white rabbits by feeding a high cholesterol diet for 12 weeks for screening of drugs against atherosclerosis. After 12 weeks, blood was collected from ear vein for evaluation of total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL) and very low density lipoprotein (VLDL) levels, then the animals were sacrificed to collect the livers for estimation of cholesterol, and aorta for gross and histopathological evaluations. The elevated levels of serum and liver parameters accompanied by gross and histopathological changes like accumulation of foam cells, atheromatous plaque formation and replacement fibrosis supported the successful induction of atherosclerosis in New Zealand white rabbits.

The protective effect of HD-03 in CCl4-induced hepatic encephalopathy in rats.

The liver is a major parenchymal organ involved in many functional activities in the body. Hepatic encephalopathy is a syndrome characterized by increased blood ammonia level and is one of the major complications of cirrhosis. In the present study the protective effect of HD-03, a poly-herbal formulation, was evaluated against CCl4-induced hepatic encephalopathy in rats. Hepatic encephalopathy was induced in Wistar rats by administration of CCl4 at a dose of 1 mL/kg orally in liquid paraffin (1:1) twice a week for 90 days. The liver enzymes (SGPT and SGOT) and blood ammonia levels were significantly (p < 0.001) higher in the CCl4-intoxicated group compared with the untreated control group. Administration of HD-03 at a dose of 750 mg/kg orally as an aqueous suspension significantly prevented the elevation of SGPT, SGOT and blood ammonia levels. Histomorphometric evaluation of liver and brain showed a protective effect of the HD-03 treatment, thus correlating with the changes in biochemical profiles. The protective effect of HD-03 against CCl4-induced encephalopathy may be due to the improved hepatocellular function, which in turn helps in regulating the metabolism of ammonia. However, further studies are required to measure the activity of enzymes involved in the urea cycle and brain aromatic amino acids in order to elucidate the exact mechanism of action of HD-03.

The beneficial effect of OST-6 (OsteoCare), a herbomineral formulation, in experimental osteoporosis.

OST-6 (OsteoCare), a herbomineral formulation, was evaluated for its inhibitory effect on the progress of bone loss induced by ovariectomy in rats. Ovariectomized (Ovx) rats were administered with OST-6 at 250 and 500 mg/kg b.wt., orally daily for 90 days. On 91st day, ovariectomized rats showed reduced bone mineral content and increased serum alkaline phosphatase levels, excretion of urinary calcium and pyridinium cross-links levels. Histologically, bone sections revealed narrowed and disappearance of trabeculae and widened medullary spaces. The total numbers of Tartrate-resistant acid phosphatase (TRAP) positive cells were significantly increased both in-vivo and in-vitro methods. OST-6, at a dose of 500 mg/kg, significantly improved bone mineral contents, serum alkaline phosphatase levels, reduced the elevated urinary calcium and pyridinium cross-links excretion, number of TRAP positive cells and reversal of the above mentioned histological features. These results indicate the usefulness of OST-6 in the management of osteoporosis in a natural way through herbal resources.

Bone mineralization by OST-6 (OsteoCare), a herbomineral preparation, in experimentally induced rickets in rats.

In the present study the efficacy of OST-6 (OsteoCare), a herbomineral preparation, on bone mineralization in experimental rickets has been evaluated. This was accomplished by feeding pregnant rats and subsequently their pups with vitamin D and calcium deficient (VDCD) with low phosphorus diet. The parameters such as serum and bone mineral contents (calcium and inorganic phosphorus), serum alkaline phosphatase, sex hormones and histology of bone were considered. VDCD resulted in a significant reduction in bone and serum calcium and inorganic phosphorus, increased serum alkaline phosphatase and decreased sex hormones (testosterone in males, progesterone and oestrogen in females). Histologically the bone showed osteodystrophic changes and disproportionate cartilaginous proliferations in the epiphyseal region. Incorporation of OST-6 into feed at 5% concentration resulted in a complete reversal of rickets, which was substantiated by biochemical and histological observations. It has been concluded that OST-6 is useful in the management of rickets in a natural way through herbal resources.

Effect of HD-03--a herbal formulation in galactosamine-induced hepatopathy in rats.

The effect of HD-03 a herbal preparation was studied on galactosamine (400 mg/kg b.wt., i.p.) induced hepatotoxicity in rats. Animals were pre-treated for 14 days with HD-03 and compared against untreated group for SGPT, SGOT, serum bilirubin and liver glycogen. Histopathology of liver lobes was considered to evaluate the extent of hepatic injury induced by galactosamine. These were reversed by HD-03 pre-treatment. HD-03 provided convincing evidence of hepatoprotection against galactosamine induced hapatotoxicity.

Anti-inflammatory, antioxidant and antimicrobial activity of Ophthacare brand, an herbal eye drops.

In the present study, the herbal preparation of Ophthacare brand eye drops was investigated for its anti-inflammatory, antioxidant and antimicrobial activity, using in vivo and in vitro experimental models. Ophthacare brand eye drops exhibited significant anti-inflammatory activity in turpentine liniment-induced ocular inflammation in rabbits. The preparation dose-dependently inhibited ferric chloride-induced lipid peroxidation in vitro and also showed significant antibacterial activity against Escherichia coli and Staphylococcus aureus and antifungal activity against Candida albicans. All these findings suggest that Ophthacare brand eye drops can be used in the treatment of various ophthalmic disorders.

Inhibition of experimental cirrhosis in rats by HD-03.

AIM: To investigate the protective effect of HD-03 in experimental cirrhosis following chronic intoxication with thioacetamide (TAA). METHODS: The effect of HD-03 (750 mg/kg p.o.) was studied in rats following TAA-induced intoxication (50 mg/kg p.o.) for a period of 90 d. HD-03 was administered as an aqueous suspension. Levels of biochemical markers indicative of hepatotoxicity were assessed in serum and liver. Histopathological evaluation of liver was also carried out to find out the protective effect of HD-03 following TAA-induced chronic intoxication. RESULTS: Administration of TAA at a dose of 50 mg/kg p.o. for 90 d resulted in a significant derangement of serum [serum glutamic pyruvate transaminase (SGPT), serum glutamic oxaloacetate transaminase (SGOT), alkaline phosphatase (ALP), albumin and bilirubin] and hepatic (triglycerides, protein, hydroxyproline, collagen and glycogen) biochemical parameters. Histopathological evaluation of liver sections following TAA-intoxication showed necrosis and proliferative changes characteristic of cirrhosis. Simultaneous treatment of TAA-intoxicated rats with HD-03 at a dose of 750 mg/kg p.o. for the same duration significantly prevented the changes in both serum and hepatic biochemical parameters. The reversal of serum and hepatic biochemical parameters also correlated with the preservation of liver histoarchitecture in HD-03 treated rats. CONCLUSION: The responses such as membrane stabilization, hepatocellular regeneration, and inhibition of collagen formation are the contributing factors in the correction of TAA-induced cirrhosis by HD-03.

Pharmacokinetic interactions of Mentat with carbamazepine and phenytoin.

In the present study, Mentat, a herbomineral psychotropic preparation, was studied for its pharmacokinetic interaction with the commonly used anti-epileptic drugs, carbamazepine and phenytoin. The interaction of carbamazepine and phenytoin with Mentat was studied in rabbits. Thirty two rabbits were divided into four groups of eight each. Animals of Group I were treated with carbamazepine (50 mg/kg b.wt. p.o.), Group II were treated with carbamazepine (50 mg/kg b.wt. p.o.) + Mentat (500 mg/kg b.wt. p.o.), Group III were treated with phenytoin (50 mg/kg b.wt. p.o.) and Group IV were treated with phenytoin (50 mg/kg b.wt. p.o.) + Mentat (500 mg/kg b.wt. p.o.) for a period of 8 days. On day 0 and day 8, plasma carbamazepine and phenytoin levels were estimated at different time intervals. A simultaneous treatment with Mentat resulted in a significant increase in plasma AUC of carbamazepine as well as phenytoin as compared to carbamazepine or phenytoin alone. Cmax and Tmax of carbamazepine and phenytoin also were evaluated. The results suggest that co-administration of Mentat could improve the effectiveness of anti-epileptic drugs due to the increased bioavailability of the latter. However, this has to be done with critical medical supervision to avoid any toxic reactions and preferably with therapeutic drug monitoring (TDM) which could also help in dose optimization.

Pharmacokinetic interaction of Diabecon (D-400) with rifampicin and nifedipine.

In the present study, Diabecon (D-400), a herbomineral anti-diabetic preparation, was studied for its pharmacokinetic interaction with the commonly used drugs rifampicin and nifedipine. Interaction of Diabecon with rifampicin: The pharmacokinetic interaction of rifampicin and Diabecon (D-400) was studied in animal models as well as in healthy human volunteers. Twelve rabbits were divided into two groups of six each. Animals in group I were treated with rifampicin (100 mg/kg body weight, p.o.) and group II with rifampicin (100 mg/kg body weight, p.o.) and Diabecon (D-400) (1 g/kg body weight, p.o.) for a period of 8 days. Rifampicin levels in plasma were estimated on day 1 and day 8 at 2, 4, 6 and 8 h after drug administration. On the basis of these findings, a clinical study in 9 healthy human volunteers aged 25-35 years and weighing 50-75 kg was initiated. They were given 450 mg of rifampicin once only on day 1 and from the second day onwards were given 2 tablets of Diabecon (D-400) twice daily for 7 days. On day 9, another dose of rifampicin (450 mg) was given along with 2 tablets of Diabecon (D-400). Blood samples were collected at 2, 4, 6 and 8 h after drug administration on day 1 and day 9 to estimate the rifampicin levels in plasma. Interaction of Diabecon with nifedipine: In another study, 12 rabbits were divided into two groups of 6 each. Group I animals were treated with nifedipine (2.5 mg/kg body weight, p.o.) and Group II animals were treated with nifedipine (2.5 mg/kg body weight, p.o.) and Diabecon (D-400) (1 g/kg body weight, p.o.) for a period of 8 days. On day 1 and day 8, blood samples were collected at 1, 2, 4 and 6 h after drug administration and plasma nifedipine levels were estimated. The results of these three studies revealed that Diabecon (D-400) did not alter the pharmacokinetic profiles of rifampicin and nifedipine.

Antioxidant activity of AO-8, a herbal formulation in vitro and in vivo experimental models.

The effect of AO-8, a herbal formulation was evaluated for free radical scavenging activity using in vitro and in vivo experimental models. AO-8 dose dependently inhibited ferric ion induced lipid peroxidation in vitro at 125-1000 micrograms. AO-8 was investigated at dose levels of 250, 500 and 750 mg/kg p.o. in isoproterenol-induced myocardial infarction and CCl4-induced hepatotoxicity in rats. The levels of serum GOT and LDH, cardiac glutathione, SOD and catalase were estimated following induction of myocardial infarction with isoproterenol. The estimation of parameters such as SGOT, SGPT, liver glycogen and lipid peroxidation were carried out in CCl4-induced hepatotoxicity. Treatment with AO-8 for 15 days at a dose of 500 and 750 mg/kg offered marked protection in both in vivo models. The reversal of increased serum enzymes in both the models may be due to the prevention of leakage of the intracellular enzymes by its membrane stabilizing activity. The inhibition of lipid peroxidation and enhancement of antioxidant enzymes by AO-8 may be due to the direct free radical scavenging activity which could be attributed to the antioxidant potential of various ingredients present in the formulation. Thus it can be concluded that AO-8 is an effective free radical scavenger and could prove beneficial in the treatment of various disorders associated with the involvement of free radicals.

Protective effect of Prostane in experimental prostatic hyperplasia in rats.

AIM: Prostane, a polyherbal formulation, was evaluated for its efficacy on 5alpha-reductase inhibition, alpha-adrenergic antagonistic activity and testosterone-induced prostatic hyperplasia. METHODS: 5alpha-reductase inhibition was evaluated using rat prostate homogenate as an enzyme source. Adrenergic antagonistic activity was evaluated using isolated rat vas deferens. Experimental prostatic hyperplasia was induced in rats by giving testosterone 3 mg/kg sc for 21 days. RESULTS: Prostane dose-dependently inhibited 5alpha-reductase activity and exhibited alpha-adrenergic antagonistic activity. Treatment with Prostane at 250, 500 and 750 mg/kg body wt, po for 21 days significantly reduced the prostatic weight, the epithelial height and the stromal proliferation in experimental prostatic hypertrophy. CONCLUSION: Prostane is effective in the treatment of experimental prostatic hypertrophy in rats and may be passed on to clinical trials on benign prostatic hypertrophy after necessary toxicological evaluations.

Anti-cholestatic activity of HD-03, a herbal formulation in thioacetamide (TAA)-induced experimental cholestasis.

In the present study HD-03, a herbal formulation was investigated for its anti-cholestatic activity in TAA-induced cholestasis in anaesthetized guinea pigs. Administration of TAA at a dose of 100 mg/kg body wt significantly reduced the bile flow, bile acid and bile salt excretion. Pretreatment with HD-03 at a dose of 750 mg/kg body wt per orally for 15 days in guinea pigs significantly prevented thioacetamide-induced changes in bile flow, bile acids and bile salts excretion. Thus, HD-03 can serve as a potent choleretic and anti-cholestatic agent.

Protective effect of HD-03, a herbal formulation, against various hepatotoxic agents in rats.

HD-03 is a polyherbal formulation containing plant drugs which are known for their hepatoprotective properties in the Ayurvedic system of medicine. In the present study, the formulation was evaluated for its protective effect against diverse hepatotoxic agents viz., paracetamol, thioacetamide and isoniazid. Treatment with HD-03 led to significant amelioration of toxin-induced changes in the biochemical parameters. Since the protective effect of HD-03 was observed in all three types of intoxication, which are different in their primary mechanism of inducing hepatotoxicity, a protective mode of action of HD-03, not specific to the hepatotoxin, is suggested.

Evaluation of possible mechanism of anti-ulcerogenic activity of UL-409, a herbal preparation.

UL-409, a herbal preparation was investigated for its anti-ulcerogenic activity and possible mechanism of action in different experimental models. Oral administration of UL-409 at a dose of 600 mg/kg significantly reduced the volume of gastric secretion, total acidity and free acidity in aspirin + pylorus-ligated rats. The drug appeared to strengthen the gastric mucosal defense mechanism by significantly increasing the total carbohydrate:protein ratio in aspirin + pylorus-ligated rats. The major mechanism involved appears to be due to promotion of mucosal protection by augmenting gastric mucin activity. Pre-treatment with UL-409 showed inhibition of alcohol-induced contraction of isolated rat fundus preparation which was reversed by indomethacin suggesting a possible involvement of the cyclo-oxygenase system.

Interaction of a herbomineral preparation D-400, with oral hypoglycaemic drugs.

D-400, a herbomineral preparation has proven antidiabetic activity in experimental models as well as in clinical trials. The possibility of concomitant use of this drug with sulphonylureas was explored in animal models. D-400 has been investigated for its interaction with oral hypoglycaemic agents namely, tolbutamide and glibenclamide in alloxan-induced diabetic rabbits. Administration of D-400 at a dose of 1 g/kg for 15 days significantly elevated plasma tolbutamide and glibenclamide concentrations with simultaneous reduction of blood glucose. Plasma tolbutamide and glibenclamide concentrations were significantly lowered after withdrawal of D-400 treatment. Elevation of plasma concentration of tolbutamide was observed only for the first 4 h after which it declined towards normal levels and no significant difference between D-400 treated and control group was observed at the end of 8 h. Significant elevation of plasma glibenclamide levels was observed at 2, 4 and 8 h with D-400 treatment. Incubation of D-400 with tolbutamide in plasma resulted in a significant increase in free tolbutamide levels.

Effect of a herbomineral preparation D-400 in streptozotocin-induced diabetic rats.

This study was undertaken to investigate the effect of D-400, a herbomineral formulation on blood sugar and other biochemical parameters in streptozotocin-induced diabetic rats. There was significant reduction in blood urea nitrogen, serum creatinine and serum uric acid levels in the D-400-treated group and considerable lowering of AUC in oral glucose tolerance test (OGTT). The D-400-treated group showed significant lowering of triglycerides and HDL cholesterol. There was a rise in hepatic glycogen level close to normal after D-400 treatment. In the pancreas of diabetic rats, D-400 therapy showed significant increase in islet number and beta cell count and appeared to bring about blood sugar homoeostasis by increasing insulin secretion and repair/regeneration of endocrine pancreas.