Prakash Chandra Bhatt

Umbelliferone β-D-galactopyranoside inhibits chemically induced renal carcinogenesis via alteration of oxidative stress, hyperproliferation and inflammation: possible role of NF-κB

Umbelliferone (7-hydroxycoumarin) possesses strong anti-inflammatory properties and free radical scavenging activity. The intent of the current study was to examine the renal protective efficacy of Umbelliferone β-D-galactopyranoside (UFG) against oxidative stress, inflammation and renal injury in Wistar rats, initiated by diethylinitrosamine (DEN) and promoted by ferric nitrilotriacetate (Fe-NTA). The capacity of UFG to scavenge the reactive nitrogen species (RNS) and reactive oxygen species (ROS) was evaluated and was also scrutinized for its effectiveness in scavenging the hydroxyl (OH), superoxide (O2), nitric acid (NO) and hydrogen peroxide (H2O2) radicals. Renal carcinoma was induced by a single intraperitoneal injection of DEN (200 mg kg−1 bodyweight) and promoted by twice weekly treatment of Fe-NTA (9 mg kg−1) for 22 weeks. To estimate the molecular mechanism involved in the antitumor potential of UFG, its effect on renal tumor inflammation was evaluated; the proinflammatory cytokines included interleukin-1β (IL-1β), interlukin-6 (IL-6) and tumor necrosis factor (TNF-α); the inflammatory mediator included prostaglandin E2 (PGE2), ornithine decarboxylase (ODC) and nuclear factor kappa B cell (NFκB). Serum abnormalities, including creatinine, lactate dehydrogenase (LDH), blood urea nitrogen (BUN) and [3H] thymidine incorporation were also induced. Furthermore, renal lipid peroxidation (LPO), endogenous antioxidant enzymes, phase II metabolizing enzymes and concomitant reduction in glutathione (GSH) were augmented. UFG showed the 95% and 99% antioxidant activity in the 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) models. UFG significantly inhibited the RNS and ROS radical and indicated the antioxidant activity (in vitro). The results showed momentous renal markers and oxidative stress protection impaired by UFG. UFG also restored the altered inflammatory and proinflammatory cytokines, which further strengthens the renal protection of UFG in DEN + Fe-NTA induced renal carcinogenesis. These results indicate that UFG is an efficient chemoprotective agent, having the ability to thwart DEN induced and Fe-NTA promoted renal carcinoma in experimental rats.

Scopolamine induced behavioral and biochemical modifications and protective effect of Celastrus paniculatous and Angelica glauca in rats

Introduction: Nootropic agents, including cholinesterase inhibitors are being used to improve memory, mood and behavior, but the side-effects associated with these agents have made their use limited. The present study has therefore been undertaken to assess the synergistic effects of Celastrus paniculatous and Angelica glauca on scopolamine induced dementia in rats. Materials and Methods: Rats were treated with scopolamine (1 mg/kg body weight, i.p.) alone and with donepezil (2 mg/kg body weight p.o.), C. paniculatous (150 mg/kg body weight, p.o) and A. glauca (150 mg/kg body weight, p.o.). The changes in behavioral and biochemical parameters were assessed in rats. Results: Scopolamine treated rats showed impaired learning and memory, increased activity of acetylcholinesterase (AChE) (18%), lipid peroxidation (60%), protein carbonyls (47%) and decreased levels of reduced glutathione (GSH) (35%), activity of superoxide dismutase (34%) and catalase (42%) in hippocampus as compared with control. Simultaneous treatment of C. paniculatous and A. glauca with scopolamine also caused an improvement in the learning and memory activity associated with AChE activity in hippocampus of rats as compared to those treated with scopolamine alone. Combined treatment of C. paniculatous, A. glauca and scopolamine significantly improved the learning and memory function and AChE activity (30%) associated with decreased lipid peroxidation (33%), protein carbonyls (27%) and increased levels of antioxidant enzymes like reduced GSH (46%), activity of superoxide dismutase (50%) and catalase (62%) in hippocampus of rats as compared with those treated with scopolamine alone. Conclusion: The results of the present study exhibit protective efficacy of combined treatment of C. paniculatous and A. glauca in scopolamine induced dementiaand promising as a memory enhancing agents that is associated with its strong antioxidant potential.

Nose to brain delivery of astaxanthin-loaded solid lipid nanoparticles: fabrication, radio labeling, optimization and biological studies

The present study was carried out to investigate the intranasal delivery of astaxanthin as solid lipid nanoparticles with the intention of improving brain targeting of astaxanthin for neurological disorders. The astaxanthin solid lipid nanoparticles were prepared by a double emulsion solvent displacement method. Statistical analysis using response surface methodology showed that the optimum values of stearic acid (50 mg), drug weight percentage (6.11%) and ratio of surfactant to co-surfactant (poloxamer 188 : lecithin-1 : 6) resulted in a 213.23 nm particle size and 0.367 polydispersity index for the astaxanthin solid lipid nanoparticles. Radio labeling studies were performed using technetium-99m to evaluate the biodistribution pattern after administration through different routes in experimental subjects. Radiolabeled nanoparticles were found to be 96–98% stable even after 48 h of labeling in phosphate-buffered saline (pH 7.4). Comparative biodistribution data indicated that the higher drug concentration in the brain was achieved by intranasal administration of 99mTc labeled astaxanthin solid lipid nanoparticles as compared to the intravenous route, which was also confirmed by gamma scintigraphy analysis. Furthermore, studies on the pheochromocytoma-12 cell line demonstrated the antioxidant potential of astaxanthin solid lipid nanoparticles against H2O2 induced toxicity. Our findings strongly emphasize that nanoparticle-based nasal drug delivery of astaxanthin could impart the utmost neuroprotection from oxidative stress in neurological disorders under in vitro conditions.

Attenuation of neurobehavioral and neurochemical abnormalities in animal model of cognitive deficits of Alzheimer’s disease by fermented soybean nanonutraceutical

The present study was performed to evaluate the efficacy of nanonutraceuticals (NN) for attenuation of neurobehavioral and neurochemical abnormalities in Alzheimer’s disease. Solid-state fermentation of soybean with Bacillus subtilis was performed to produce different metabolites (nattokinase, daidzin, genistin and glycitin and menaquinone-7). Intoxication of rats with colchicine caused impairment in learning and memory which was demonstrated in neurobehavioral paradigms (Morris water maze and passive avoidance) linked with decreased activity of acetylcholinesterase (AChE). NN treatment led to a significant increase in TLT in the retention trials as compared to acquisition trial TLT suggesting an improved learning and memory in rats. Further, treatment of NN caused an increase in the activity of AChE (42%), accompanied with a reduced activity of glutathione (42%), superoxide dismutase (43%) and catalase (41%). It also decreased the level of lipid peroxidation (28%) and protein carbonyl contents (30%) in hippocampus as compared to those treated with colchicine alone, suggesting a possible neuroprotective efficacy of NN. Interestingly, in silico studies also demonstrated an effective amyloid-β and BACE-1 inhibition activity. These findings clearly indicated that NN reversed colchicine-induced behavioral and neurochemical alterations through potent antioxidant activity and could possibly impart beneficial effects in cognitive defects associated with Alzheimer’s disease.

Fabrication, optimization, and characterization of umbelliferone β-D-galactopyranoside-loaded PLGA nanoparticles in treatment of hepatocellular carcinoma: in vitro and in vivo studies

Umbelliferone β-D-galactopyranoside (UFG), isolated from plants, exhibits promising inhibitory action on numerous diseases. The present research was initiated to develop a suitable delivery system for UFG with an intention to enhance its therapeutic efficacy against diethyl nitrosamine (DEN)-induced hepatocellular carcinoma (HCC) in Wistar rats. UFG-loaded polymeric nanoparticles prepared by sonication were scrutinized for average size, drug loading capacity, zeta potential, and drug release potency in animals. HCC cell lines HuH-7 and Hep G2 were used for in vitro cytotoxic investigation. Several hepatic, nonhepatic, antioxidant, and anti-inflammatory biochemical parameters were estimated to establish the anticancer potential of UFG nanoformulation. Microscopical and histopathological investigations were also undertaken to substantiate the results of our work. Umbelliferone β-D-galactopyranoside-loaded poly(d,l-lactide-co-glycolide) nanoparticles (UFG-PLGA-NP) with particle size of 187.1 nm and polydispersity index 0.16 were uniform in nature with 82.5% release of the total amount of drug after 48 h. Our study successfully established the development and characterization of UFG-PLGA-NP with noticeable effect against both in vivo and in vitro models. The anticancer potential of UFG-PLGA-NP was brought about by the management of DEN-induced reactive oxygen species generation, mitochondrial dysfunction, proinflammatory cytokines alteration, and induction of apoptosis. Positive zeta potential on the surface of UFG-PLGA-NP would have possibly offered higher hepatic accumulation of UFG, particularly in the electron-dense mitochondria organelles, and this was the take-home message from this study. Our results demonstrated that such polymer-loaded delivery systems of UFG can be a better option and can be further explored to improve the clinical outcomes against hepatic cancer.

Umbelliferon-α-d-glucopyranosyl-(2I → 1II)-α-Dglucopyranoside ameliorates Diethylnitrosamine induced precancerous lesion development in liver via regulation of inflammation, hyperproliferation and antioxidant at pre-clinical stage

It is well documented that anomalous production of inflammatory proteins linked with most of the toxic expression and genesis of diverse chronic disease including cancer. Diethylnitrosamine (DEN) a well-known hepatotoxin and hepatocarcinogen, can induce oxidative stress and inflammatory reaction in it. Umbelliferone, secondary metabolites, is present in different plants and widely consumed by humans as medicine and food supplements. The aim of the current study was to scrutinize the chemoprotective potential of umbelliferon-α-d-glucopyranosyl-(2I→1II)-α-d-glucopyranoside (UFD) against DEN-induced hepatocellular carcinoma (HCC) in experimental rats. Single intraperitoneal injection of DEN (200mg/kg) was used for induction of HCC in rats and rats were grouped and orally treated with UFD (5, 10 and 20mg/kg) dose for 22 weeks. Parameters under investigation included hepatic, non-hepatic enzymes, oxidative stress, pro-inflammatory cytokines, COX-2 and NF-κB level along with histopathological examination in HCC rats. UFD exerted protective effect via reduction of oxidative stress, liver and non-liver parameters in a dose-dependent manner. It also reduced the expression of TNF-α, IL-1β, IL-6 and COX-2 in diseased rats. Our result revealed the essential repression of the inflammation cascade through modulation of nuclear factor-kappa B (NF-κB) signaling pathway.

Influence of high-pressure homogenization, ultrasonication, and supercritical fluid on free astaxanthin extraction from β-glucanase-treated Phaffia rhodozyma cells

ABSTRACT In this study astaxanthin production by Phaffia rhodozyma was enhanced by chemical mutation using ethyl methane sulfonate. The mutant produces a higher amount of astaxanthin than the wild yeast strain. In comparison to supercritical fluid technique, high-pressure homogenization is better for extracting astaxanthin from yeast cells. Ultrasonication of dimethyl sulfoxide, hexane, and acetone-treated cells yielded less astaxanthin than β-glucanase enzyme-treated cells. The combination of ultrasonication with β-glucanase enzyme is found to be the most efficient method of extraction among all the tested physical and chemical extraction methods. It gives a maximum yield of 435.71 ± 6.55 µg free astaxanthin per gram of yeast cell mass.

Development of surface-engineered PLGA nanoparticulate-delivery system of Tet-1-conjugated nattokinase enzyme for inhibition of Aβ 40 plaques in Alzheimer’s disease

According to the World Health Organization, globally there are around 18 million patients suffering from Alzheimer’s disease (AD), and this number is expected to double by 2025. The pathophysiology of AD includes selective deposition of Aβ peptide in the mitochondria of cells, which inhibits uptake of glucose by neurons and key enzyme functions. Current drug treatments for AD are unable to rectify the underlying pathology of the disease; they only provide short-term symptomatic relief, so there is a need for the development of newer treatment regimes. The antiamyloid activity, antifibrinolytic activity, and antithrombotic activity of nattokinase holds potential for the treatment of AD. As nattokinase is a protein, its stability restricts its usage to a greater extent, but this limitation can be overcome by nanoencapsulation. In this work, we successfully synthesized polymeric nanoparticles of nattokinase and characterized its use by different techniques: transmission electron microscopy, scanning electron microscopy, DTS Nano, differential scanning calorimetry, Fourier-transform infrared spectroscopy, thioflavin T-binding assay, in vitro drug release, antifibrinolytic activity, and in vivo antiamyloid activity. As brain targeting of hydrophilic drugs is complicated due to the stringent nature of blood–brain barrier, in the current experimental study, we conjugated poly(lactic-co-glycolic acid) (PLGA)-encapsulated nattokinase with Tet1 peptide, which exhibits retrograde transportation properties because of its affinity to neurons. Our study suggests that PLGA-encapsulated nattokinase polymeric nanoparticles are able to downregulate amyloid aggregation and exhibit antifibrinolytic activity. The encapsulation of nattokinase in PLGA did not affect its enzyme activity, so the prepared nanoformulation containing nattokinase can be used as an effective drug treatment against AD.

Commentary: L-3-n-butylphthalide Rescues Hippocampal Synaptic Failure and Attenuates Neuropathology in Aged APP/PS1 Mouse Model of Alzheimer's Disease

Mouse Model of Alzheimers Disease. According to World Health Organization estimation about 18 million people worldwide are suffering from Alzheimers disease (AD) and figures would shoot up to 34 million by 2025 in aging population. The onset and propagation of AD is unrelenting from cellular to molecular levels, governed by multitude of factors leading to a continual decrease in cognitive abilities (Felsenstein et al., 2014). Recent advances in understanding the genetic factors that predispose to AD, as well as in biomarker development have brought with them the promise of earlier and more reliable diagnosis of this disease. However, treatment regime and strategies available, approved by US-FDA are limited by their scope to alter pharmacological targets that can provide symptomatic relief to the patient (Lannfelt et al., 2014). The present stock of drugs for AD consists of donepezil, galantamine, rivastigmine, and memantine. The primary mechanism of action for these drugs is to decrease the acetyl cholinesterase (AChE) levels thus altering acetylcholine level, while memantine is a low-affinity voltage-dependent uncompetitive antagonist at glutamatergic N-methyl d-aspartate receptor (NMDA) thereby blocking the activity of the neurotransmitter glutamate. All the existing treatments for AD are only symptomatic but non curative (Lannfelt et al., 2014). Under these circumstances a drug molecule with multiple pharmacological properties with intent to modify as well as reduce the risk factors contributing toward disease burden will be highly appreciated. Drugs that are aimed at disrupting AD disease progression by inhibition/degradation of the protein mis-folding of β-amyloid (Aβ) into neurotoxic oligomeric aggregates has been encouraged for AD (Scheff et al., 2007). With the advent of new diagnostic tools various pathological conditions have been identified at different stages of AD. Accumulation of amyloid beta (Aβ) to form clumps known as neuritic or senile plaques is considered as hallmark of AD. The buildup of these plaques seems to be a crucial as they lead to oxidative stress coupled with mitochondrial dysfunction and barrage of inflammatory cascade causing neuronal cell death. Studies carried by Zhang et al. (2016) on L-3-n-butylphthalide

Preclinical renal chemo-protective potential of Prunus amygdalus Batsch seed coat via alteration of multiple molecular pathways

Abstract Prunus amygdalus Batsch (almond) is a classical nutritive traditional Indian medicine. Along with nutritive with anti-oxidant properties, it is, clinically, used in the treatment of various diseases with underlying anti-oxidant mechanism. This study is an effort to scrutinise the renal protective effect of P. amygdalus Batsch or green almond (GA) seed coat extract and its underlying mechanism in animal model of Ferric nitrilotriacetate (Fe-NTA) induced renal cell carcinoma (RCC). RCC was induced in Swiss Albino Wistar rats by intraperitoneal injection of Fe-NTA. The rats were then treated with ethanolic extract of GA (25, 50 and 100 mg/kg per oral) for 22 weeks. Efficacy of GA administration was evaluated by change in biochemical, renal, macroscopical and histopathological parameters and alterations. Additionally, interleukin-6 (IL-6), tumour necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and inflammatory mediator including prostaglandin E2 (PGE2), nuclear factor-kappa B (NF-κB) were also observed to explore the possible mechanisms. The oral administration of GA significantly (p < .001) altered the Fe-NTA induced RCC in rats by inhibition of renal nodules, decolourisation of tissues, tumour promoter marker including thymidine 3[H] incorporation, ornithine decarboxylase, renal parameters and anti-oxidant parameters in serum. Additionally, GA treatment significantly (p < .001) down-regulated the IL-6, IL-1β, TNF-α, inflammatory mediators PGE2 and NF-κB in a dose-dependent manner. Histopathology observation supported the renal protective effect of GA by alteration in necrosis, size of Bowman capsules and inflammatory cells. Hence, it can be concluded that GA possesses observable chemo-protective action and effect on Fe-NTA induced RCC via dual inhibition mechanism one by inhibiting free radical generation and second by inhibiting inflammation.