Gollapalle Lakshminarayanashastry Viswanatha

An Improved Experimental Model of Hemorrhoids in Rats: Evaluation of Antihemorrhoidal Activity of an Herbal Formulation

Objective. To improve the existing experimental model of croton oil-induced hemorrhoids in rats by using Evans Blue (EB) dye extravasation technique. Further, an herbal formulation (Pilex) was evaluated for its antihemorrhoidal activity in this model. Methods. Two sets of experiments were carried out: first to improve the experimental model and to validate the same using Pilex and second to evaluate the effect of Pilex on cytoarchitecture of rectoanal tissue in croton oil-induced hemorrhoids. In both sets, hemorrhoids were induced to all the animals, except normal controls, by applying croton oil via rectoanal region and the effect of Pilex ointment (PO), Pilex granules (PG), and combination of PG and PO was evaluated. In the first set, extravasation of EB dye, TNF-α, IL-6, and rectoanal coefficient (RAC) was determined. In the second set, severity of score, RAC, and histopathology were evaluated. Results. The elevated levels of TNF-α, IL-6, and extravasations of EB dye were decreased with the Pilex treatment. The cytoarchitecture of rectoanal portion of the animals treated with Pilex was near to normal. Conclusion. The improved experimental model of hemorrhoid is useful in quantifying the inflammatory exudates and extent of inflammation. In this improved experimental model Pilex showed antihemorrhoidal activity, which further validates its clinical usage.

Poly-Ingredient Formulation Bresol® Ameliorates Experimental Chronic Obstructive Pulmonary Disease (COPD) in Rats

In the present study, the protective effect of Bresol® – a polyherbal formulation – was evaluated in an experimental model of cigarette smoke (CS)-induced COPD in rats. Ten minutes daily exposure to CS for 7 weeks caused significant elevation of TNF-α (p<0.01) and total protein (p<0.01) in the bronchoalveolar lavage fluid (BALF) of positive untreated control animals, indicating ongoing inflammatory process in the lungs. Further, histopathological findings have confirmed the presence of pathological lesions in the trachea and lungs. Five weeks of post-treatment with Bresol® (250 and 500 mg/kg, p.o.) showed significant and dose-dependent anti-inflammatory effects against CS-induced lung abnormalities by maintaining the TNF-α and total protein levels within the normal range. Additionally, Bresol®-treated animals showed normal cyto-architecture of the trachea and lungs. In conclusion, Bresol® showed dose-dependent protection against CS-induced lung and tracheal injury in rats, which further indicates, Bresol® is a useful healing agent, may help to decelerate the progression of COPD, and reduce the exacerbations in patients.

LC-MS/MS profiling and neuroprotective effects of Mentat® against transient global ischemia and reperfusion-induced brain injury in rats.

OBJECTIVE The aim of this study was to evaluate the possible beneficial effects of Mentat against transient global ischemia and reperfusion-induced brain injury in rats. METHODS The neuroprotective effects of Mentat were evaluated against transient global ischemia and reperfusion (I/R)-induced brain injury in rats. Various neurobehavioral and biochemical parameters were assessed, followed by morphologic and histopathologic evaluation of brain tissue to conclude the protective effect of Mentat. Additionally, in vitro antioxidant assays were performed to explore the antioxidant capacity of Mentat and detailed liquid chromatography-mass spectrometry (LC-MS/MS) profiling was carried out to identify the active phytoconstituents responsible for the protective effects of Mentat. RESULTS Sixty minutes of transient global ischemia followed by 24 h reperfusion (I/R) caused significant alterations in the cognitive and neurologic functions in the ischemia control group (P < 0.01) compared with the sham control. Furthermore, 2,3,5-triphenyltetrazolium chloride staining of the ischemia control group showed 20.85% ± 0.39% of cerebral infarct area (P < 0.01), increased brain volume (% edema 17.81% ± 1.576%; P < 0.01), and increased lipid peroxidation (P < 0.01) in the brain homogenate. Additionally, the histopathology of the ischemia control group showed severe brain injury compared with the sham control group. Interestingly, pretreatment with Mentat (250 and 500 mg/kg, p.o.) and quercetin (20 mg/kg, p.o.) for 7 d has alleviated all pathological changes observed due to I/R injury. Mentat also showed very good antioxidant activity in in vitro assays (2,2-diphenyl-l-picrylhydrazyl, ferric-reducing antioxidant power, and oxygen radical absorbance capacity assays). Furthermore, the detailed LC-MS/MS analysis of Mentat was performed and enclosed for identifying the actives responsible for its protective effects. CONCLUSIONS These findings suggest that Mentat is a neuroprotective agent that may be a useful adjunct in the management of ischemic stroke and its rehabilitation especially with respect to associated memory impairment and other related neurologic conditions.

Novel experimental model of non-infectious pharyngitis in rats.

INTRODUCTION Currently, there is a paucity of scientific literature and reports related to screening models for non-infectious type of pharyngitis. In this context, we made a sincere attempt to establish a novel animal model for screening drugs against non-infectious pharyngitis in rats. We have considered the use of pyridine, croton oil and their combination for inducing non-infectious pharyngitis in rats. METHODS Various concentrations of pyridine were applied topically to the pharyngeal region of rats and the extent of inflammation was assessed by Evans Blue (EB) dye exudation test, evaluating the serum levels of proinflammatory cytokines and histopathology. Dexamethasone and diclofenac were used as reference standards. RESULTS Upon pyridine application (2.5%, 5%, 10%, 20%, 40% and 80% in saline), dose-dependent increase in EB dye extravasation was observed (increased vascular permeability). In addition, the levels of TNF-α (P<0.01) and IL-6 (P<0.01) were significantly increased compared to control. Furthermore, the histopathology of pharyngeal tissue showed hypertrophy of submucosal glands, severe inflammation of the pharynx characterised by presence of mononuclear cells, neutrophils along with haemorrhages and congestion; however, normal control animals showed normal cytoarchitecture of pharynx. Indeed, dexamethasone (0.25, 0.5 and 1 mg/kg, i.v.) and diclofenac (1, 2.5 and 5 mg/kg, i.v.) showed dose-dependent protection against pyridine-induced pharyngitis. Further, the possible mechanism of pyridine-induced pharyngitis is thought to be primarily mediated through phospholipase A2 and cyclooxygenase (COX) pathway. CONCLUSION These findings suggest that pyridine-induced pharyngitis is a simple and versatile novel animal model for screening the drugs against non-infectious pharyngitis in rats.

Anti-platelet and Anti-thrombotic Effects of a Poly-ingredient formulation: In vitro and in vivo experimental evidences

OBJECTIVE The present study was conducted to evaluate the efficacy of Abana® (a poly-ingredient formulation with natural constituents) on in vitro platelet aggregation and occlusion-induced deep venous thrombosis in rats. METHODS Anti-platelet property of Abana® was evaluated using ADP (Adenosin 5-diphosphate) and adrenaline-induced platelet aggregation models, and anti-thrombotic activity was evaluated against occlusion-induced deep venous thrombosis model in wistar rats. RESULTS Under the in vitro conditions, Abana® (250, 500 and 1000 µg/ml) alleviated ADP and adrenaline-induced platelet aggregation in a dose-dependent manner. Abana® (1000 µg/ml) inhibited ADP and adrenaline-induced platelet aggregation by as much as 50.69% and 64.83% respectively. Furthermore, 6 days pre-treatment with Abana® (250 and 500 mg/kg, p.o.) in an in vivo study showed significant and dose-dependent protection against occlusion-induced deep venous thrombosis in rats. CONCLUSION These findings suggest that Abana®, a polyherbal formulation possesses anti-platelet and anti-thrombotic activities in the experimental models of in vitro platelet aggregation and in vivo deep venous thrombosis in rats.

IM-133N - A Useful Herbal Combination for Eradicating Disease-triggering Pathogens in Mice via Immunotherapeutic Mechanisms

Objectives: The present study was undertaken to evaluate the immunomodulatory (IM) activity of IM-133N, a herbal combination in various immunotherapeutic experimental models. Methods: The IM activity of IM-133N was evaluated against three experimental models namely, effect of IM- 133N against Escherichia coli (E. coli)-induced abdominal sepsis in mice, and carbon clearance test was performed in Wistar albino rats to evaluated the phagocytic potential of IM-133N, in addition IM-133N was evaluated for its immunoglobulin enhancing potential in rats, where the immunoglobulin levels were measured by zinc sulphate turbity (ZST) test. Further, IM-133N was subjected for detailed liquid chromatography-mass spectrometry (LC-MS)/MS analysis to identify the probable active constituents present in it. Results: The findings of the present study has demonstrated very promising IM property of IM-133N in all the experimental models. Briefly, pretreatment with IM-133N at 125, 250, 500 and 1,000 mg/kg, p.o. doses had protected the mice against E. coli-induced abdominal sepsis and mortality, further the effect of IM- 133N was found to be significant and dose-dependent. In support of this, in another study administration of IM-133N showed a significant and dose-dependent increase in serum immunoglobulin levels, estimated by ZST test. In line with the above findings, in the carbon clearance test the low doses (125 and 250 mg/ kg, p.o.) of IM-133N increased the rate of carbon clearance, whereas the higher doses (500 and 1,000 mg/kg, p.o.) did not sustain the response, and saturation effect was considered as one of the possible reason for futility of higher doses for IM-133N. In addition, A detailed LC-MS/MS analysis of IM-133N showed 17 bioactive phytochemical constituents: namely, apigenin, chaulmoogric acid, mesquitol, quercetin, symphoxanthone, salireposide, β-sitosterol, nonaeicosanol, β-amyrin, betulic acid, oleanolic acid, symplososide, symponoside, symploveroside, symplocomoside, symconoside A and locoracemoside B. Conclusion: These findings suggest that IM-133N possesses significant IM activity and, hence, could be useful for eradicating opportunistic disease-triggering pathogens via immunotherapeutic mechanisms. The findings also suggest IM-133N may also useful in other immunity disorders.

Ameliorative effect of Koflet formulations against pyridine-induced pharyngitis in rats

In present study two formulations of Koflet (syrup and lozenges) were evaluated against pyridine-induced pharyngitis in rats. Topical application of 10% pyridine showed extravasation of Evans blue stain as a characteristic feature of on-going inflammation. In addition, the levels of TNF-α (p < 0.01) and IL-6 (p < 0.01) were significantly increased compared to control. Further, histopathology of the pharyngeal tissue showed submucosal gland hypertrophy, severe mucosal inflammation characterized by presence of mononuclear cells and neutrophils along with haemorrhages and congestion; however, saline applied animals (normal control) showed normal cytoarchitecture of the pharynx. Interestingly, pre-treatment with dexamethasone (1 mg/kg, p.o.), Koflet lozenges (KL) (500 and 1000 mg/kg, p.o.) and Koflet syrup (KS) (2 and 4 ml/kg, p.o.) for 7 days showed significant and dose dependent protection by decreasing the EB dye extravasation, and serum levels of TNF-α and IL-6. In addition, histopathological findings have further supported the protective effect of Koflet formulations. These findings suggest that, both Koflet syrup and Koflet lozenges are highly effective in treating non-infectious type of pharyngitis. Among the two formulations KS was found to be more potent than KL, and possible mechanism of action thought to be mediating through inhibition of TNF-α and/or phospholipids–arachidonic acid pathway.