Jitendra K. Rawat

Alpha-linolenic acid stabilizes HIF-1 α and downregulates FASN to promote mitochondrial apoptosis for mammary gland chemoprevention

Alpha linolenic acid is an essential polyunsaturated fatty acid and is reported to have the anti-cancer potential with no defined hypothesis or mechanism/s. Henceforth present study was in-quested to validate the effect of alpha linolenic acid on mitochondrial apoptosis, hypoxic microenvironment and de novo fatty acid synthesis using in-vitro and in-vivo studies. The IC50 value of alpha linolenic acid was recorded to be 17.55μM against ER+MCF-7 cells. Treatment with alpha linolenic acid was evident for the presence of early and late apoptotic signals along with mitochondrial depolarization, when studied through acridine orange/ethidium bromide and JC-1 staining. Alpha linolenic acid arrested the cell cycle in G2/M phase. Subsequently, the in-vivo efficacy was examined against 7, 12-dimethylbenz anthracene induced carcinogenesis. Treatment with alpha linolenic acid demarcated significant effect upon the cellular proliferation as evidenced through decreased in alveolar bud count, restoration of the histopathological architecture and loss of tumor micro vessels. Alpha linolenic acid restored the metabolic changes to normal when scrutinized through 1H NMR studies. The immunoblotting and qRT-PCR studies revealed participation of mitochondrial mediated death apoptosis pathway and curtailment of hypoxic microenvironment after treatment with alpha linolenic acid. With all above, it was concluded that alpha linolenic acid mediates mitochondrial apoptosis, curtails hypoxic microenvironment along with inhibition of de novo fatty acid synthesis to impart anticancer effects.

Effect of enteral administration of α-linolenic acid and linoleic acid against methotrexate induced intestinal toxicity in albino rats

The present study was conducted to show the effect of α-linolenic acid (ALA) (18 : 3, ω-3) and linoleic acid (LA) (18 : 2, ω-6) on experimental intestinal toxicity induced by methotrexate (MTX). The groups of albino rats received: Group I: normal saline (2 ml kg−1, i.p. sham control); Group-II: MTX (2.5 mg kg−1, i.p. toxic control); Group-III: ALA (2 ml kg−1, i.p.); Group-IV: LA (18 : 2, ω-6) (2 ml kg−1, i.p.), Group-V: ALA (2 ml kg−1, i.p.) and Group-VI: LA (2 ml kg−1, i.p.) with MTX (2.5 mg kg−1, i.p.). Animals were sacrificed after 7 days treatment schedule and appraised for intestinal pH, total acidity, free acidity and colonic mucosal disease index (CMDI). Intestinal tissues were further evaluated for oxidative stress parameters (TBARS, SOD, protein carbonyl and catalase), and morphological modulation using scanning electron microscopy. The intestinal tissues were further graded for the enzymatic activities of COX-1, COX-2 and 15-LOX. Both ALA and LA demonstrated momentous protection against MTX induced intestinal toxicity, which could be attributed to their prooxidant nature.

Efficacy of variable dosage of aspirin in combating methotrexate-induced intestinal toxicity

The aim of the present study was to study in detail the effect of variable doses of aspirin on intestinal toxicity. Albino rats were randomly divided into six groups and subjected to 13 weeks treatment against a sham control (3 ml kg−1 by mouth (p.o.) normal saline); a toxic control (2.5 ml kg−1 intraperitoneal injection (i.p.), MTX); a low dose of aspirin (8 mg kg−1, p.o.); a low dose of aspirin plus MTX (8 mg kg−1, p.o. + 2.5 ml kg−1, i.p.), a high dose of aspirin (45 mg kg−1, p.o.); and a high dose of aspirin plus MTX (45 mg kg−1, p.o. + 2.5 ml kg−1, i.p.). The intestinal toxicity of aspirin was assessed on the basis of biochemical changes and modulation in the inflammatory markers. Low doses of aspirin gave significant protection against MTX-induced toxicity, whereas high dose failed to do so. High doses of aspirin also produced adverse biochemical changes in the physiology, but low dose did not.

Effect of rutin against gastric esophageal reflux in experimental animals

Abstract Context: The present study was undertaken to elucidate the effect of rutin against gastric esophageal reflux in experimental animals. Methods: Groups of rats, fasted overnight received normal saline (3 ml/kg, sham control) or esophagitis control (3 ml/kg, normal saline) or pantoprazole (30 mg/kg) or rutin (50 and 100 mg/kg) were subjected to pylorus and forestomach ligation. Animals were sacrificed after 12 h and scrutinized physiologically (gastric pH, total acidity, free acidity and esophagitis index), biochemically (TBAR’s, SOD, catalase, GSH and protein carbonyl) and morphologically. The esophageal tissues were also inquested for the presence of proinflammatory (IL-2 and IL-1β) and immunoregulatory (IL-4 and IL-6) cytokines. Results: The results demonstrated momentous physiological, biochemical and morphological protection imparted by rutin. The rutin also restored the altered levels of proinflammatory and immunoregulatory cytokines, which further strengthens the implication of rutin in GERD. Conclusion: The beneficial effects as observed in the current experiment could be accredited to the antioxidant and anti-inflammatory (through inhibition of COX and LOX) property of rutin.

Rifaximin, a pregnane X receptor (PXR) activator regulates apoptosis in a murine model of breast cancer

The present study was proposed to investigate the effect of rifaximin (RFX) on methyl nitrosourea (MNU) induced mammary gland carcinoma in albino wistar rats. Animals were randomized and divided among four groups of six animals each. Group I (control 0.9% normal saline, 3 ml kg−1, p.o.); Group II (toxic control, MNU 47 mg kg−1, i.v.); Group III (RFX, 25 mg kg−1, p.o.); Group IV (RFX, 50 mg kg−1, p.o.). Toxicity was induced by single i.v. injection of MNU. MNU treatment was evident with increased alveolar bud count, differentiation score, up-regulated inflammatory enzyme markers (COX, LOX, NO and H2S) and oxidative stress markers (TBARs, protein carbonyl, SOD, catalase and Ach). The mammary gland surface architecture was studied using SEM, carmine staining and H&E staining. The treatment with RFX elicited noticeable restoration of the overall histological architecture in the experimental animals similar to the control. In the MNU treated toxic group, the levels of oxidative stress markers significantly increased in comparison to the control, which was subsequently restored after RFX treatment. Furthermore, RFX up regulated the levels of caspase 3 and caspase 8, when compared to the MNU treated animals. MNU associated toxicity was also ascertained, when determined for UCHL-1, COX, NF-κBp65, BAD, and BCL-xl expression, while RFX demonstrated modulation of the same.

Comparative efficacy of alpha-linolenic acid and gamma-linolenic acid to attenuate valproic acid-induced autism-like features

The present study was undertaken to elucidate the effect of alpha-linolenic acid (ALA, 18:3, ω-3) and gamma-linolenic acid (GLA, 18:3, ω-6) on experimental autism features induced by early prenatal exposure to valproic acid (VPA) in albino wistar pups. The pups were scrutinized on the accounts of behavioral, biochemical, and inflammatory markers, and the results suggested that the GLA can impart significant protection in comparison to ALA against VPA-induced autism features. When scrutinized histopathologically, the cerebellum of the GLA-treated animals was evident for more marked protection toward neuronal degeneration and neuronal loss in comparison to ALA. Concomitant administration of ALA and GLA with VPA demonstrated a marked cutdown in the Pgp 9.5 expression with GLA having more pronounced effect. Henceforth, it can be concluded that ALA and GLA can impart favorable protection against the VPA-induced autism-like features with GLA having pronounced effect.

Preclinical appraisal of terbutaline analogues in precipitation of autism spectrum disorder

Terbutaline is a β2 agonist used in the clinical management of asthma and as a tocolytic agent during pregnancy. In the recent past, the preterm use of terbutaline has been shown to hasten autistic-like symptoms in offsprings. For this reason, the present study was carried out to understand the effects of pharmacological analogues of terbutaline (salbutamol, salmeterol and montelukast) in the progression of ASDs in experimental animals. Pregnant rats were treated with salbutamol (10 mg kg−1, sc), salmeterol (10 μg kg−1, sc) and montelukast (10 mg kg−1, sc) and the offsprings were scrutinized for behavioral, biochemical, neuro-inflammatory and histopathological changes. The offsprings from the rats treated with salbutamol and montelukast were determined to be closely associated with various symptoms of ASDs.

GLA supplementation regulates PHD2 mediated hypoxia and mitochondrial apoptosis in DMBA induced mammary gland carcinoma

The aim of the present study is to evaluate the effect of gamma linolenic acid (GLA) on mitochondrial mediated death apoptosis, hypoxic microenvironment and cholinergic anti-inflammatory pathway against 7, 12-dimethylbenz (a) anthracene (DMBA) induced mammary gland carcinoma. The effects of GLA were evaluated morphologically and biochemically against DMBA induced mammary gland carcinoma. The metabolic study was done for evaluation of biomarkers using 1H NMR. The present study was also verified through immunoblotting and qRT-PCR studies for the evaluation of various pathways. GLA treatment has a delineate implementation upon morphology of the tissues when evaluated through carmine staining, hematoxyline and eosin staining and scanning electron microscopy. GLA also demarked a commendatory proclamation of the fifteen key serum metabolites analogous with amino acid metabolism and fatty acid metabolism when recognized through1H NMR studies. The immunoblotting and qRT-PCR studies accomplished that GLA mediated mitochondrial death apoptosis, curtail hypoxic microenvironment along with hindrance of de novo fatty acid synthesis and also mediate the cholinergic anti-inflammatory pathway to proclaim its anticancer effects.

Palonosetron attenuates 1,2-dimethyl hydrazine induced preneoplastic colon damage through downregulating acetylcholinesterase expression and up-regulating synaptic acetylcholine concentration

The present study was undertaken to evaluate the effect of palonosetron (PAL) against 1,2-dimethylhydrazine (DMH)-induced colon cancer. Wistar albino rats were randomly divided into four groups (n = 8). Group 1 served as normal control (1 mM EDTA + saline, 2 ml per kg per day, s.c.); group 2 toxic control; group 2, 3 and 4 received DMH (20 mg per kg per week, s.c.), for 6 weeks; groups 3 and 4 also received PAL (0.25 and 0.50 mg per kg per day, p.o) for 6 weeks. DMH treated rats showed altered heart rate variability (HRV) factors, increased incidence of aberrant crypt foci (ACF), distorted antioxidant markers (TBARs, SOD, catalase, GSH) and increased levels inflammatory markers (COX). The colonic surface architecture, studied using scanning electron microscopy (SEM), revealed aberrant crypts (500×) and preneoplastic nodules (2000×). PAL treatment helped to minimize the ACF count, and restored oxidative and inflammatory markers favorably. To further validate our results, we directed our study to define the effect of PAL on acetylcholine (Ach) neurotransmission using a simple model organism, C. elegans. Increased cholinergic transmission by PAL (8 μM) was evident in the worms when studied through an aldicarb assay. However, PAL (2 μM, 4 μM and 8 μM) treatment negatively modulated nAchR, when evaluated using the levamisole assay. The increased synaptic Ach levels can be attributed to the decreased levels of acetylcholinesterase (AchE), which could be attributed to the decreased genomic levels of ace-1 and ace-2. The above findings were also supported by the fact, that we observed decreased AchE activity in PAL treated rats. In addition the downregulation in the expression of unc-38 (one of the necessary components of nAchR) sufficiently links with the decreased nAchR activity. Our findings emphasize the potential role of PAL in the suppression of colon carcinogenesis.

Effects of minocycline and doxycycline against terbutaline induced early postnatal autistic changes in albino rats

The current study was initiated to explicate the shielding response of minocycline and doxycycline against early postnatal neurological damage and behavioral alteration convinced by terbutaline. Toxicity was induced by terbutaline at three successive days in the pups. The pups were scrutinized for behavioral, biochemical and inflammatory markers. Subsequent treatment with test drugs commenced a favorable effect on the autistic symptoms with more safeguard by doxycycline. The study also recognized peripheral inflammatory reactions and increased nitric oxide (NO) through terbutaline which was curtailed down by test drugs, with the much more noticeable effect of doxycycline. The GC-FID analysis and histopathological evaluation of the brain tissue elicited more pronounced protection by doxycycline. Doxycycline was also evident with remarkable down-regulation Pgp 9.5 [Ubiquitin carboxy-terminal hydrolase L1 (UCHL-1)] expression in the brain tissue in comparison to minocycline.