Presence of pyrroloquinazoline alkaloid in Adhatoda vasica attenuates inflammatory response through the downregulation of pro-inflammatory mediators in LPS stimulated RAW 264.7 macrophages
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 inf...
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| Format: | Article |
| Language: | English |
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Tabriz University of Medical Sciences
2021-01-01
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| Series: | BioImpacts |
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| Online Access: | https://bi.tbzmed.ac.ir/PDF/bi-11-15.pdf |
| _version_ | 1819098574412054528 |
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| author | Reddy Amala Sundaresan Sujatha |
| author_facet | Reddy Amala Sundaresan Sujatha |
| author_sort | Reddy Amala |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-22T00:33:09Z |
| format | Article |
| id | doaj.art-5e6e5b79a73d49e6bc3d37e918434834 |
| institution | Directory Open Access Journal |
| issn | 2228-5660 2228-5652 |
| language | English |
| last_indexed | 2024-12-22T00:33:09Z |
| publishDate | 2021-01-01 |
| publisher | Tabriz University of Medical Sciences |
| record_format | Article |
| series | BioImpacts |
| spelling | doaj.art-5e6e5b79a73d49e6bc3d37e9184348342022-12-21T18:44:53ZengTabriz University of Medical SciencesBioImpacts2228-56602228-56522021-01-01111152210.34172/bi.2021.03bi-21844Presence of pyrroloquinazoline alkaloid in Adhatoda vasica attenuates inflammatory response through the downregulation of pro-inflammatory mediators in LPS stimulated RAW 264.7 macrophagesReddy Amala0Sundaresan Sujatha1Animal Cell Culture Laboratory, Department of Biotechnology, SRMIST, Kattankulathur, Tamilnadu, IndiaAnimal Cell Culture Laboratory, Department of Biotechnology, SRMIST, Kattankulathur, Tamilnadu, IndiaIntroduction: 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.https://bi.tbzmed.ac.ir/PDF/bi-11-15.pdfinducible nitric oxide synthasecyclooxygenases-2lipopolysaccharideraw 264.7 macrophagesquinazoline alkaloidsdiabetic foot ulcerlimb amputation |
| spellingShingle | Reddy Amala Sundaresan Sujatha Presence of pyrroloquinazoline alkaloid in Adhatoda vasica attenuates inflammatory response through the downregulation of pro-inflammatory mediators in LPS stimulated RAW 264.7 macrophages BioImpacts inducible nitric oxide synthase cyclooxygenases-2 lipopolysaccharide raw 264.7 macrophages quinazoline alkaloids diabetic foot ulcer limb amputation |
| title | Presence of pyrroloquinazoline alkaloid in Adhatoda vasica attenuates inflammatory response through the downregulation of pro-inflammatory mediators in LPS stimulated RAW 264.7 macrophages |
| title_full | Presence of pyrroloquinazoline alkaloid in Adhatoda vasica attenuates inflammatory response through the downregulation of pro-inflammatory mediators in LPS stimulated RAW 264.7 macrophages |
| title_fullStr | Presence of pyrroloquinazoline alkaloid in Adhatoda vasica attenuates inflammatory response through the downregulation of pro-inflammatory mediators in LPS stimulated RAW 264.7 macrophages |
| title_full_unstemmed | Presence of pyrroloquinazoline alkaloid in Adhatoda vasica attenuates inflammatory response through the downregulation of pro-inflammatory mediators in LPS stimulated RAW 264.7 macrophages |
| title_short | Presence of pyrroloquinazoline alkaloid in Adhatoda vasica attenuates inflammatory response through the downregulation of pro-inflammatory mediators in LPS stimulated RAW 264.7 macrophages |
| title_sort | presence of pyrroloquinazoline alkaloid in adhatoda vasica attenuates inflammatory response through the downregulation of pro inflammatory mediators in lps stimulated raw 264 7 macrophages |
| topic | inducible nitric oxide synthase cyclooxygenases-2 lipopolysaccharide raw 264.7 macrophages quinazoline alkaloids diabetic foot ulcer limb amputation |
| url | https://bi.tbzmed.ac.ir/PDF/bi-11-15.pdf |
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