Drug Standardization through Pharmacognostic Approaches and Estimation of Anticancer Potential of Chamomile (Matricaria chamomilla L.) using Prostate-Cancer cell lines: An In-vitro Study.

Cancer is the major challenge across world and the adenocarcinoma of prostate malignancy is the second most prevalent male cancer. Various medicinal plants are used for the treatment and management of various cancers. Matricaria chamomilla L., is one of the extensively used Unani medicament for the treatment of various type of diseases. In the current study we evaluated most of the parameters prescribed for drug standardization using pharmacognostic approaches. The 2,2 Diphenyl-1-picryl hydrazyl (DPPH) method was utilized for the analysis of antioxidant activity in the flower extracts of M. chamomilla. Moreover, we analyzed the antioxidant and cytotoxic activity of M. chamomilla (Gul-e Babuna) through in-vitro method. DPPH (2,2-diphenyl-1-picryl-hydrazl-hydrate) method was utilized for the analysis of antioxidant activity in the flower extracts of M. chamomilla. CFU and wound healing assay were performed to determine the anti-cancer activity. The results demonstrated that various extracts of M. chamomilla fulfilled most of the parameters of drug standardization and contained good antioxidant and anticancer activities. The ethyl acetate showed higher anticancer activity followed by aqueous, hydroalcoholic, petroleum benzene and methanol by CFU method. Also, the wound healing assay demonstrated that ethyl acetate extract has more significant effect followed by methanol and petroleum benzene extract on prostate cancer cell line (C4-2). The current study concluded that the extract of M. chamomilla flowers could act as good source of natural anti-cancer compounds.

Independent Association of 25[OH]D Level on Reduced Glutathione and TNF-α in Patients with Diabetes and/or Hypertension.

Purpose: Oxidative and inflammatory pathways play a significant role in the pathophysiology of a wide variety of non-communicable diseases such as type 2 diabetes mellitus (T2DM) and hypertension. However, the effect of serum 25-hydroxyvitamin D (25[OH]D) on these pathways is still controversial. To evaluate the association of 25[OH]D on antioxidant and pro-inflammatory biomarkers, reduced glutathione (GSH) and tumor necrosis factor (TNF)-α, in T2DM and hypertensive patients. Patients and Methods: This is a cross-sectional study of a consecutive sample of patients attending the the Family Medicine clinic at King Abdullah bin Abdulaziz University Hospital (KAAUH). Participants were screened for eligibility according to the following criteria: aged above 18 years and diagnosed with T2DM and/or hypertension for at least one year. Patients receiving any kind of vitamin D or calcium supplements within the last three months were excluded, as were those with a history of renal failure, cancer, liver, thyroid, or any other chronic inflammatory diseases. Results: In total 424 T2DM and/or hypertensive patients (mean age 55±12 years) were recruited. In addition to routine physical and laboratory examinations, levels of serum 25[OH]D, GSH and TNF-α were measured. The prevalence of 25[OH]D deficiency (<50 nmol/L) was 35.1%, which was independent from GSH and TNF-α levels. In T2DM, hypertensive and patients having both diseases, GSH levels were 349.3±19, 355.4±19 and 428.8±20 µmol/L, respectively. Uncontrolled T2DM and hypertension patients showed significantly higher GSH compared with the controlled group. Males showed slightly higher level of TNF-α compared with females and uncontrolled hypertensive patients had relatively higher TNF-α level when evaluated against controlled hypertensive patients. . Conclusion: 25[OH]D level is independent of oxidative stress and inflammation, assessed by levels of GSH and TNF-α, respectively, in T2DM and hypertensive Saudi patients. .

Computational and Biological Comparisons of Plant Steroids as Modulators of Inflammation through Interacting with Glucocorticoid Receptor.

Despite the usefulness of glucocorticoids, they may cause hazardous side effects that limit their use. Searching for compounds that are as equally efficient as glucocorticoids, but with less side effects, the current study compared plant steroids, namely, glycyrrhetinic acid, guggulsterone, boswellic acid, withaferin A, and diosgenin with the classical glucocorticoid, fluticasone. This was approached both in silico using molecular docking against glucocorticoid receptor (GR) and in vivo in two different animal models. All tested compounds interacted with GR, but only boswellic acid and withaferin A showed docking results comparable to fluticasone, as well as similar in vivo anti-inflammatory effects, by significantly decreasing serum levels of interleukin-6 and tumor necrosis factor-α in cotton pellet-induced granuloma in rats. In addition, both compounds significantly decreased the percent of change in ear weight in croton oil-induced ear edema in mice and the granuloma weight in cotton pellet-induced granuloma in rats, to levels comparable to that of fluticasone. Both boswellic acid and withaferin A had no effect on adrenal index, but only withaferin A significantly increased the thymus index. In conclusion, boswellic acid may have comparable anti-inflammatory effects to fluticasone with fewer side effects.

Ginsenoside-Rb1 ameliorates lithium-induced nephrotoxicity and neurotoxicity: Differential regulation of COX-2/PGE2 pathway.

To investigate the effect of Ginsenoside-Rb1 (GRb1) on lithium (Li+)-induced toxicity, GRb1 was given to rats orally (100mg/kg) for 14days. In independent groups, lithium chloride (4meq/kg/day i.p.) was administered at day 4 of the experiment for 10days, with or without GRb1. Li+ caused significant deterioration of behavioral responses including righting reflex, spontaneous motor activity and catalepsy. Li+ also caused distortion in normal renal, cerebral and cerebellum architecture and significantly worsened all kidney functional parameters tested compared to control. In addition, Li caused oxidative stress in both kidney and brain, evident by significant increase in malondialdehyde and nitric oxide levels, with decrease in reduced glutathione and catalase activity. Administration of GRb1 prior to Li+ significantly improved behavioral responses, renal and brain histopathological picture, kidney function tests and oxidative stress markers compared to sole Li+-treated group. Concomitant administration of GRb1 decreased Li+ levels by about 50% in serum, urine and brain and by 35% in the kidney. Interestingly, Li+ had a differential effect on cyclooxygenase (COX)-2/prostaglandin E2 (PGE2) pathway, as it significantly increased COX-2 expression and PGE2 level in the kidney, while decreasing them in the brain compared to control. On the other hand, administering GRb1 with Li+ suppressed COX-2/PGE2 pathway in both kidney and brain compared to Li+ alone. In conclusion, GRb1 can alter Li+ pharmacokinetics resulting in extensively decreasing its serum and tissue concentrations. Furthermore, COX-2/PGE2 pathway has a mechanistic role in the nephro- and neuro-protective effects of GRb1 against Li+-induced toxicity.

Protective mechanisms of atorvastatin against doxorubicin-induced hepato-renal toxicity.

To investigate the mechanisms by which the anticancer drug doxorubicin (DOX)-induced hepato-renal damage could be prevented by the cholesterol-lowering statin, atorvastatin (Ator), Ator (10 mg/kg) was administered orally for 10 days, and, in independent rat groups, DOX hepato-renal toxicity was induced via a single i.p. dose of 15 mg/kg at day 5 of experiment, with or without Ator. DOX caused deterioration in hepato-renal function, as it significantly increased blood urea nitrogen (BUN), creatinine, alanine transaminase (ALT) and aspartate transaminase (AST) compared to control, with distortion in normal renal and hepatic histology. Pretreatment with Ator preserved kidney and liver function and histology. DOX caused oxidative stress as indicated by significant decrease in reduced glutathione (GSH) level and catalase activity with increase in malondialdehyde (MDA) compared to control. Combined DOX/Ator significantly reversed these values compared to DOX in both kidney and liver. DOX caused nitrosative stress, as it increased tissue nitric oxide compared to control. Concomitant DOX/Ator treatment decreased NO in kidney and liver. Furthermore, DOX caused inflammatory effects indicated by up-regulation of hepato-renal nuclear factor-κB (NF-κB) expression and increment of tumor necrosis factor-α (TNF-α) tissue concentration, with down-regulation of endothelial nitric oxide synthase (eNOS). DOX also caused apoptotic effect, as it up-regulated the apoptotic marker, Bcl-2-associated X protein (Bax), expression in liver and kidney. Using Ator with DOX reversed hepato-renal inflammatory and apoptotic marker expression. These findings suggest Ator as a protective adjuvant against DOX toxicity, via antioxidant, anti-nitrosative, anti-inflammatory and anti-apoptotic mechanisms.

Functional role of arginine 375 in transmembrane helix 6 of multidrug resistance protein 4 (MRP4/ABCC4).

Multidrug resistance protein (MRP) 4 transports a variety of endogenous and xenobiotic organic anions. MRP4 is widely expressed in the body and specifically localized to the renal apical proximal tubule cell membrane, where it mediates the excretion of these compounds into urine. To characterize the MRP4 substrate-binding site, the amino acids Phe368, Phe369, Glu374, Arg375, and Glu378 of transmembrane helix 6, and Arg998 of helix 12, localized in the intracellular half of the central pore, were mutated into the corresponding amino acids of MRP1 and MRP2. Membrane vesicles isolated from human embryonic kidney 293 cells overexpressing these mutants showed significantly reduced methotrexate (MTX) and cGMP transport activity compared with vesicles that expressed wild-type MRP4. The only exception was substitution of Arg375 with serine, which had no effect on cGMP transport but significantly decreased the affinity of MTX. Substitution of the same amino acid with a positively charged lysine returned the MTX affinity to that of the wild type. Furthermore, MTX inhibition of MRP4-mediated cGMP transport was noncompetitive, and the inhibition constant was increased by introduction of the R375S mutation. A homology model of MRP4 showed that Arg375 and Arg998 face right into the central aqueous pore of MRP4. We conclude that positively charged amino acids in transmembrane helices 6 and 12 contribute to the MRP4 substrate-binding pocket.