Neuroprotective effect of herbal extracts inhibiting soluble epoxide hydrolase (sEH) and cyclooxygenase (COX) against chemotherapy-induced cognitive impairment in mice.

Chemotherapy-induced cognitive impairment (CICI) is a novel clinical condition characterized by memory, learning, and motor function deficits. Oxidative stress and inflammation are potential factors contributing to chemotherapy's adverse effects on the brain. Inhibition of soluble epoxide hydrolase (sEH) has been proven effective in neuroinflammation and reversal of memory impairment. The research aims to evaluate the memory protective effect of sEH inhibitor and dual inhibitor of sEH and COX and compare its impact with herbal extracts with known nootropic activity in an animal model of CICI. In vitro sEH, the inhibitory activity of hydroalcoholic extracts of Sizygium aromaticum, Nigella sativa, and Mesua ferrea was tested on murine and human sEH enzyme as per the protocol, and IC50 was determined. Cyclophosphamide (50 mg/kg), methotrexate (5 mg/kg), and fluorouracil (5 mg/kg) combination (CMF) were administered intraperitoneally to induce CICI. The known herbal sEH inhibitor, Lepidium meyenii and the dual inhibitor of COX and sEH (PTUPB) were tested for their protective effect in the CICI model. The herbal formulation with known nootropic activity viz Bacopa monnieri and commercial formulation (Mentat) were also used to compare the efficacy in the CICI model. Behavioral parameter such as cognitive function was assessed by Morris Water Maze besides investigating oxidative stress (GSH and LPO) and inflammatory (TNFα, IL-6, BDNF and COX-2) markers in the brain. CMF-induced CICI, which was associated with increased oxidative stress and inflammation in the brain. However, treatment with PTUPB or herbal extracts inhibiting sEH preserved spatial memory via ameliorating oxidative stress and inflammation. S. aromaticum and N. sativa inhibited COX2, but M. Ferrea did not affect COX2 activity. Lepidium meyenii was the least effective, and mentat showed superior activity over Bacopa monnieri in preserving memory. Compared to untreated animals, the mice treated with PTUPB or hydroalcoholic extracts showed a discernible improvement in cognitive function in CICI.

Herbal Combination 'HAGE-101912' Ameliorates Gastroesophageal Reflux Disease in Rats.

Background: Gastroesophageal reflux disease (GERD) is a chronic, relapsing disorder. In this era of modern and fast-track lifestyle and food habits, the incidence of GERD is rapidly increasing. Currently, proton pump inhibitors (PPIs) are the primary choice of treatment. However, the associated side effects and a high relapse rate give rise to the need to explore alternative therapies. Objective: The study aimed to evaluate HAGE-101912, an herbal combination, in different experimental models of GERD. Methods: Antacid activity was assessed based on H+/K+ATPase inhibitory activity of parietal cells using artificial gastric juice. Tonic contraction of the lower esophageal sphincter (LES) was evaluated using an AD Instrument. A GERD model of the pylorus and fundus ligation (preventive and curative models) in rats was selected to assess the efficacy of HAGE-101912 at a dose of 250 mg/kg body weight, and various parameters such as the gastric pH, gastric volume, total acidity, gross esophageal ulcer index, and histopathological changes were evaluated. The prokinetic activity was assessed using the phenol red method. Results: HAGE-101912 increased the acid-neutralizing capacity (P < 0.001), decreased H+/K+ATPase activity (P < 0.01), and increased the contraction of the LES. In the preventive model, HAGE-101912 significantly reduced the gastric acid volume (P < 0.01), total acidity (P < 0.001), and gross esophageal ulcer index (P < 0.01); increased the gastric acid pH (P < 0.01); and protected the esophageal epithelium. In addition, HAGE-101912 increased gastric emptying and gastrointestinal transit through its prokinetic activity (P < 0.05). Conclusion: HAGE-101912 has a beneficial effect in GERD as it effectively inhibits the H+/K+ATPase, increases the gastric pH, restores the LES function, protects the esophageal epithelium, and increases gastric emptying and transit.

Investigation of HPLF-111624 in Modified Experimental Models of Ulcerative Proctitis and Anal Fissure in Rats.

Background: Although ulcerative proctitis (UP) and anal fissure (AF) are common anorectal diseases, there are no appropriate experimental models to screen the drugs intended for these conditions. In this context, existing experimental models mimicking these diseases were modified and the polyherbal formulation, HPLF-111624 was evaluated in these models. Objective: To establish animal model for UP and AF and to evaluate polyherbal formulation, HPLF-111624 in these disease models. Methods: An experimental model of UP was selected based on the modification of the ulcerative colitis model using different concentrations of acetic acid. The concentration used for induction were 2.5%, 5% and 10% v/v and different weights used to induce AF were 25 g, 50 g and 100 g, which were selected based on the severity of inflammation, fecal score, gross pathology, and histopathological evaluation. Furthermore, these animal models were used to evaluate the efficacy of HPLF-111624, a polyherbal formulation known to be beneficial in anal diseases. Results: Acetic acid at 5% produced typical pathological changes that resembled UP, with a significant increase in the fecal score, gross lesion, and histopathological changes. Similarly, among the three weights, physical injury with a 100 g weight produced significant changes in the histopathological score in the model of AF. Intervention with HPLF-111624 at doses of 250 and 500 mg/kg b.wt., showed a reduction in the inflammatory cytokines and a significant improvement in the histopathological findings in both the conditions. Conclusion: The results showed that the modified experimental models for UP and AF resemble the human pathological conditions and are simple, versatile and may be used for screening drugs intended for these conditions. Intervention with HPLF-111624 was found to be effective in improving the pathological state of UP and AF.

Pharmacological investigation of 'HIM-CHX': A herbal combination in the experimental muscle wasting condition.

Muscle wasting diseases are gradually increasing with the increase in global life expectancy. This study was designed to evaluate the efficacy of HIM-CHX, a herbal combination of Boswellia serrata, Cissus quadrangularis, and Withania somnifera, on Sarcopenia. The effects of HIM-CHX on parameters such as muscle mass, grip strength, motor coordination, gait, locomotor activity and endurance were measured in rats. In addition to this, inflammatory cytokines, myokine and growth hormone levels were also evaluated. In the first experiment, HIM-CHX was administered orally to rats at the dose of 125, 250, and 500 mg/kg body weight for 12 weeks. At the end of the treatment period, muscle mass, grip strength, motor coordination and proinflammatory cytokines were evaluated. In the second experiment, HIM-CHX was administered orally at a dose of 500 mg/kg body weight for 4 weeks and gait analysis, locomotor activity, endurance and endogenous antioxidant activity were evaluated. The animals treated with HIM-CHX showed a significant improvement in gastrocnemius muscle weight, carcass weight, gait, locomotor activity and endurance. HIM-CHX exerts its effect by reducing the levels of TNF-α, IL-6, and Myostatin while increasing the IGF-1 levels which are the typical biomarkers of muscle wasting. Furthermore, the study findings indicate that HIM-CHX has the potential to correct the pathophysiological changes associated with sarcopenia.

Effect of "DXB-2030," a Polyherbal Formulation, on Experimental Polycystic Ovary Syndrome Associated with Hyperandrogenism.

The objective of the present study was to evaluate "DXB-2030," a polyherbal combination of Trigonella foenum-graecum, Aloe vera, Sphaeranthus indicus, Nardostachys jatamansi, and Symplocos racemosa extracts in an experimental model of testosterone propionate (TP), induced polycystic ovary syndrome (PCOS) in female rats. Thirty animals were divided into 3 groups of 10 each; group 1 served as normal control; group 2 was administered with TP and served as positive control; along with TP, group 3 was treated with "DXB-2030" at a dose of 100 mg/kg p.o., for 60 days. At the end of the study period, the animals were subjected for the estimation of serum testosterone levels, oral glucose tolerance test (OGTT), weight of the ovaries, estrous cycle, and histopathological evaluation. An in vitro assay on GLUT4 expression was carried out to understand the effect of "DXB-2030" on insulin resistance. Results showed that treatment with "DXB-2030" reversed the TP-induced changes by increasing the GLUT4 expression and decreasing the body weight, testosterone levels, AUC of glucose in OGTT, and the cystic follicles of the ovaries, thus indicating its beneficial effect in PCOS by ameliorating the metabolic dysfunction and reproductive impairment, which are the pathophysiological conditions associated with PCOS. From the results obtained, it can be concluded that "DXB-2030" was effective in the management of experimental PCOS and hence may be recommended in the treatment of PCOS.

Turmeric based oral rinse "HTOR-091516" ameliorates experimental oral mucositis.

BACKGROUND: Prevalence and incidence of oral mucositis (OM) are rigorously increasing and there is no effective treatment. The herbal formulation "HTOR-091516" containing Curcuma longa, Triphala and honey were evaluated for the treatment of OM. AIM: The aim of this study was to evaluate the safety and efficacy of HTOR-091516, employing cellular model, human gingival fibroblasts-1 (HGF-1), and 5-fluorouracil (5-FU)-induced mucositis model in rats. MATERIALS AND METHODS: The cell viability was assessed using 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide (MTT) assay and the inhibitory effect of HTOR-091516 on tumor necrosis factor-alpha (TNF-α) was evaluated using TNF-α bioassay in lipopolysaccharides-induced HGF-1. 5-FU and glacial acetic acid were used to induce OM in rats. Animals were divided into two groups, group 1 served as mucositis control and group 2 was treated with HTOR-091516 at the dose of 200 µl and TNF-α was estimated in plasma samples. RESULTS: The in vitro safety of HTOR-091516 was evaluated in reconstructed human oral epidermis and was found to be nontoxic and exhibited concentration-dependent TNF-α inhibition in HGF-1. The treatment with HTOR-091516 reduced mucositis scores and mortality rate and also decreased the plasma TNF-α level. CONCLUSION: The present data indicate that HTOR-091516 is effective in the treatment of OM.

Ketosteroid Standardized Cissus quadrangularis L. Extract and its Anabolic Activity: Time to Look Beyond Ketosteroid?

BACKGROUND: Cissus quadrangularis (CQ) L. reported to contain 3-ketosteroids and have bone health benefits. AIM: This study aimed at establishing the relationship between the ketosteroid content and anabolic as well as bone health-promoting activities of various Cissus extracts in well-established orchidectomized (ORX) rat model. MATERIALS AND METHODS: Supercritical carbon dioxide, ethyl acetate, and aqueous extracts (AE) of CQ L. were prepared and standardized for ketosteroid content by two methods used in commerce. Moreover, ketosteroid standardized extracts of this plant were evaluated for anabolic activity in rats in well-established ORX rat model. RESULTS: The increase in the absolute weight was appreciable in the CQ-AE treated group. Similarly, with respect to bone parameters, a similar trend was seen. The mean bone density, strength, and calcium content were found to be highest in the group treated with CQ-AE compared to groups treated with other extracts. This study reveals for the first time that 3-ketosteroids are not linked to the beneficial activities on bone and highlights the need for extensive characterization of biological active principles from CQ L. CONCLUSION: In light of the above estimation studies, we believe that current standardization of Cissus extraction "3-ketosteroids" is incorrect. We also did not find any report suggesting the presence of androgenic steroids in this plant and hence the characterization based on "3-ketosteroids" is scientifically incorrect. This study highlights the insufficient understanding of biological active principles from CQ L. and underlines the need for extensive bioactivity guided studies. SUMMARY: Cissus quadrangularis (CQ) L. reported to contain 3.ketosteroids and have bone health benefitsWe did not find correlation between ketosteroid content obtained by conventional methods and its biological effectStudies indicate that claims of ketosteroid content need not necessarily correlate to biological effects and hence warrants extensive phytochemical characterization of biological active principles from CQ L. Abbreviations used: CQ: Cissus quadrangularis, ORX: Orchidectomized, AE: Aqueous extract, EE: Ethyl acetate extract, SFE: Supercritical fluid extract.

Extract of a polyherbal formulation ameliorates experimental nonalcoholic steatohepatitis.

The objective of the present study is to evaluate the effect of the extract of a well-known hepatospecific polyherbal formulation, Liv.52, in an experimental model of high-fat diet (HFD)-induced nonalcoholic steatohepatitis (NASH) in rats. Feeding a HFD for 15 weeks resulted in significant impairment of the lipid profile, elevation of hepatic enzyme markers, and insulin resistance in rats. The histological examination of the liver furthermore indicated fibrotic changes and fat deposition in hepatic tissues. The treatment with Liv.52 extract [125 mg/kg body weight per os (b.wt. p.o.)], which was administered from week 9 onward, reversed the HFD-induced changes to a statistically significant extent, compared to the untreated positive control animals. The effect observed with Liv.52 extract was comparable to that of pioglitazone (4 mg/kg b.wt.), a standard drug that is useful in the management of NASH. The treatment with Liv.52 extract significantly reduced steatosis, collagen deposition, and necrosis in hepatic tissues, which indicates its antifibrotic and antinecrotic properties. The results obtained in the present set of experiments indicate that Liv.52 extract effectively reverses metabolic and histological changes associated with HFD-induced NASH.

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.

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.

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.

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.

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.

Polyherbal formulation Bresol® protects the mast cells against compound 48/80-induced disruption and histamine release: a non-immunological mechanism of mast cell stabilization.

OBJECTIVE: Present study was aimed to evaluate the protective effect of Bresol®, a polyherbal formulation, on mast cell degranulation and histamine release from mast cells. METHODS: Mast cell-stabilizing activity of Bresol® was evaluated against compound 48/80-induced mast cell degranulation and histamine release from rat peritoneal mast cells in ex vivo conditions. RESULTS: Microscopy of the control group smears showed more of intact mast cells, with very minimum number of degranulated mast cells and negligible amount of histamine release. In contrast, incubation of mast cells with compound 48/80 caused significant degranulation of the mast cells associated with release of high concentration of histamine in the positive control group. Furthermore, Bresol® at 100 mg/L showed a significant inhibition of compound 48/80-induced mast cell degranulation. In addition, Bresol® significantly and dose-dependently inhibited compound 48/80-induced histamine release. CONCLUSION: Bresol® inhibits compound 48/80-induced mast cell degranulation and histamine release in ex vivo conditions. The present findings could be one of the non-immunological mechanism responsible for usefulness of Bresol® in various allergic conditions.

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.