Presence of pyrroloquinazoline alkaloid in Adhatoda vasica attenuates inflammatory response through the downregulation of pro-inflammatory mediators in LPS stimulated RAW 264.7 macrophages

Abstract

Introduction: Inflammation is the primary response caused due to harmful stimuli which are followed by the increased draining of plasma and immune cells from the body into the site of the injured tissue. A signaling cascade of growth factors and cytokines propagates and eventually matures in the inflammatory site involving the blood vessels and immune markers within the injured tissue in order to promote the renewal of the degenerated tissue. During a chronic disorder like diabetic foot ulcer, there is an obstinate inflammation which may act as a prime factor for limb amputation and upon persistent prevalence may even lead to death. Methods: This study focuses on the mode of action of ALK-F (alkaloid fraction) isolated from Adhatoda vasica in attenuating the nitric oxide production which was estimated by Griess assay, tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) expression was analyzed by ELISA and expression of COX-2 and iNOS by RT-PCR and western blotting in LPS stimulated RAW 264.7 macrophages. Total intracellular ROS was analyzed by DCFH-DA probing and the presence of quinazoline alkaloid (vasicine) in the ALK-F was evidenced by high performance liquid chromatography (HPLC). Results:The ALK-F of A. vasica exhibited a significant inhibitory effect on LPS elicited nitrite production (13.2 ± 1.06 µM), iNOS, and COX-2 (2.6 and 3.3 fold) in a dose-dependent manner. There was a significant decrease in the generation of these pro-inflammatory cytokines TNF-α (1102 ± 1.02 pg/mL) and IL-6 (18 ± 0.87 ng/mL) and total intracellular ROS in the highest tested concentrations (1 µg and 10 µg) of ALK-F of A. vasica. HPLC analysis by the gradient elution method revealed the presence of 12% of quinazoline alkaloid vasicine in the crude alkaloid fraction. Conclusion: Thus this study communally suggests that attenuation of nitric oxide and the dysregulation of genes responsible for inflammation which deliberates A. vasica to conflict against inflammation and provide remedial benefits in diabetic wound care.

Anti-inflammatory and neuroprotective activity of Viphyllin a standardized extract of β-caryophyllene from black pepper (Piper Nigrum L) and its associated mechanisms in mouse macrophage cells and Human Neuroblastoma SH-SY5Y cells

“…Furthermore, iNOS and COX-2 enzymes play crucial role in the generation of NO and PGE2, Viphyllin significantly subsided the iNOS and COX-2 expression in LPS stimulated macrophages. Our results are consistent with the previous published reports [20,21].…”

Efficacy of a Probiotic and Herbal Supplement in Models of Lung Inflammation

“…The alkaloid fraction from vasaka shows antiin ammatory effects through the TNF-α pathway, and curcumin isolated from turmeric may downregulate NF-κB, IL-1, and IL-6. 31,36 Mouse models are useful for exploring mechanisms of action but are limited in accurately modeling the multi-factorial dynamics of a human microbiome. Future studies exploring the action of the gut-lung axis and the role of probiotics will be best examined in humans.…”

“…30 Furthermore, the consumption of herbal extracts of holy basil leaf, turmeric root, and vasaka leaf shows potential in supporting bronchodilatory, antioxidant, and antiin ammatory function through the gut-lung axis. [31][32][33] In this study, we hypothesized that a probiotic with antioxidant herbal extracts could attenuate neutrophilic in ammation in vitro and in animal models.…”

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“…LPS interacts with TLR4 to produce NO, causing inflammation imbalances. iNOS gene-targeted antiinflammatory drugs are important for treating diabetic wounds and Alzheimer's 14 .…”