Johanna Rajkumar

Effect of ambrex (a herbal formulation) on oxidative stress in hyperlipidemic rats and differentiation of 3T3-L1 preadipocytes

Background: Ambrex is a polyherbal formulation which consists of Withania somnifera, Orchis mascula, Cycas circirnalis, Shorea robusta with amber. Objective: The present study was designed to explore the potential effects of ambrex on the antioxidant status in high fat diet fed rats and to investigate the possible mechanisms focusing on the gene expression involved in adipogenesis and inflammation in 3T3-L1 cell line. Materials and Methods: Male Wistar rats were divided into four groups (n = 6); Group A received normal diet, Group B received high fat diet for 30 days, Group C and D received high fat diet for 30 days and treated with ambrex (40 mg/kg b.w) and atorvastatin (10 mg/kg b.w) for successive 15 days respectively. This study also assesses the effect of ambrex on adipogenesis in 3T3-L1 adipocytes. Results: The serum total cholesterol and triglycerides were significantly decreased in ambrex treated hyperlipidemic animals when compared to untreated animals. The activities of catalase, superoxide dismutase and reduced glutathione were significantly augmented in the serum, liver, and heart of hyperlipidemic rats treated with ambrex when compared to control. Ambrex treated rats had significant reductions in malondiadehyde levels in the serum, liver and heart compared to untreated rats. In addition, we observed that treatment with ambrex resulted in a major inhibition of pre-adipocyte differentiation of 3T3-L1 cells in vitro by suppression of peroxisome proliferator activated receptor gamma, sterol regulatory binding proteins, tumor necrosis factor-α, inducible nitricoxide synthase, leptin, and upregulation of thioredoxin 1 (TRX1) and TRX2 mRNA expression. Conclusion: Therefore, ambrex may be a potential drug for treatment of hyperlipidemia and related disorders.

Hepatoprotective Activity of Heptoplus on Isoniazid and Rifampicin Induced Liver Damage in Rats.

The present study is designed to evaluate the efficacy of heptoplus a polyherbal formulation as an oral supplementary agent for isoniazid and rifampicin induced hepatotoxicity in rats. 50 and 100 mg/kg of heptoplus supplement were fed orally to the rats along with isoniazid and rifampicin and compared to rats treated with 100 mg/kg Liv 52 standard drug. Rats treated with isoniazid and rifampicin suffered from severe oxidative stress by the virtue of free radicals induced lipid per oxidation. As a result abnormal index of serum biochemical markers for liver function and increased liver lysosomal enzymes activity was observed. However rats nourished with 100 mg/kg of heptoplus and Liv 52 protected the liver from oxidative damage by maintaining normal antioxidant profile status and restored normal serum liver biochemical markers. Increased liver lysosomal enzymes activity is prevented in the rats supplemented with heptoplus and Liv 52. Histopathological analysis also revealed severe vascular changes and lobular necrosis in the treatment of isoniazid and rifampicin. Heptoplus (100 mg/kg) and Liv 52 supplemented rats liver apparently revealed normal architecture of liver. This study confirms that heptoplus has liver protective activity against Isoniazid and Rifampicin induced liver injury in rats, in par with Liv 52.

Inhibition of PKCβ Mediates Cardioprotective Activity of Ambrex against Isoproterenol-Induced Myocardial Necrosis: in vivo and in silico Studies

Aims and objectives: The current study characterized the morphology of Ambrex formulation by Scanning Electron Microscopy and assessed its cardioprotective activity against Isoproterenol (ISPH)-induced myocardial necrosis in rats by biochemical and histopathological evaluations, and also attempted to predict the prospective protein-targets of Ambrex and the signaling pathway that mediates this activity through molecular docking approach. Materials and methods: Sprague–Dawley male rats (4 groups, 6 rats per group) chosen for the current study were acclimatized to the laboratory conditions for 7 days prior to actual treatment; they were pretreated with Ambrex (40 mg/kg b.wt/day, p.o) everday for 21 days and then intoxicated with ISPH (85 mg/kg b.wt, s.c) on day-20 and 21 to experimentally induce myocardial necrosis. The extent of ISPH-induced myocardial necrosis was quantified in terms of the serum levels of two cardiac biomarkers: creatine kinase-MB and lactate dehydrogenase. The extent of ISPH-induced oxidative stress was quantified in terms of the tissue levels of five oxidative stress biomarkers: superoxide dismutase, catalase, reduced glutathione, glutathione peroxidase and lipid peroxidation. Results and discussion: The Scanning Electron Microscopy image of Ambrex formulation showed the formation of nanoparticles with thickness of 65 nm, making Ambrex a unique metal-deficient Siddha-medicine based polyherbal nano-formulation characterized and evaluated in India. Pretreatment with Ambrex attenuated the extent of ISPH-induced oxidative stress, lipid peroxidation and generation of reactive oxygen species as reflected by biochemical evaluations, and also ameliorated the degree of ISPH-induced myocardial necrosis and membrane damage as reflected by histopathological evaluations. The results of molecular docking revealed that Withaferin-A and Methyl Commate-A (the key metabolites of Withania somnifera and Ambrex respectively) inhibit Protein KinaseC Beta, and renders Ambrex its cardioprotective activity by maintaining the intracellular antioxidant homeostasis and myocardial membrane architecture.

Antimicrobial activity of a polyherbal formulation Ambrex

The present investigation was undertaken to evaluate the antimicrobial potential of Ambrex, a polyherbal formulation, against distinct microorganisms. Ambrex is a mixture of four herbs namely Withania somnifera, Orchis mascula, Pistacia lentiscus, Cycas circinalis with amber (a resin from Pinus succinifera). The aqueous extract of ambrex showed significant antimicrobial activity against Salmonella enterica, Salmonella typhi, Salmonella paratyphi, Shigella Flexneri, Pseudomonas putida and Candida albicans. In the antimicrobial well diffusion assay, substantial inhibition was observed with Salmonella typhi and Candida albicans. The antibacterial activity of Ambrex was comparable with that of the standards cifran and streptomycin. It also showed potent inhibition of the Candida Albicans and the effect is comparable to fluconazole. In summary, the present study demonstrated the potential antimicrobial activity of Ambrex, a polyherbal formulation.

STUDIES ON ACUTE TOXICITY OF HEPTOPLUS IN EXPERIMENTAL RATS

Objective: To evaluate acute toxicity of Heptoplus (polyherbal drug) in Sprague-Dawley rats and to identify the active ingredients of the herbal drug. Methods: Heptoplus was subjected to preliminary phytochemical screening and the HPTLC fingerprint profile of herbal drug was documented. OECD guideline 423-Acute toxic class method was followed to evaluate the oral toxicity of Heptoplus in Sprague-Dawley rats. In acute toxicity studies, Group I (control) rats received 0.5% of carboxymethyl cellouse (Vehicle). Group II rats received 2000 mg/kg b. w of Heptoplus. The rats were observed on the day of dosing and thereafter for 14 d, for any toxic effect. Results: Preliminary phytochemical analysis of Heptoplus revealed total phenol, flavonoid, carbohydrate, and tannins as its major constituents. The total phenol and flavonoid content of Heptoplus was found to be 170 μg of gallic acid and 162 μg of quercetin equivalent. HPTLC analysis proved that phyllanthin is an active compound of Heptoplus. Acute oral toxicity assays showed Heptoplus administration did not result in any treatment-related mortality, abnormal clinical signs, and loss of body weight or gross pathological changes in rats. Hence, LD 50 value of Heptoplus was found to be greater than 2000 mg/kg b. wt. Conclusion: Heptoplus contain phyllanthin as an active ingredient. LD 50 value of Heptoplus was found to be greater than 2000 mg/kg b. wt.