Evaluation of Cissus quadrangularis extracts as an inhibitor of COX, 5-LOX, and proinflammatory mediators
Graphical abstract
Introduction
Cissus quadrangularis is an ancient medicinal plant that has been used in many traditional medicines. Traditionally it is used to treat diverse diseases like rheumatic diseases, allergies, skin diseases, piles, fracture, and bone diseases (Udupa et al., 1965, Deka et al., 1994). It is an active ingredient of Ayurvedic formula called “Laksha Gogglu” which is used for alleviating pain, reducing inflammation and promoting the healing of wounds or simple fracture (Kulkarni, 2001). Stem of Cissus quadrangularis is very important part of the plant and used as raw drug in Indian folk medicine (Panda, 1990). Leaves of Cissus quadrangularis is used to treat anorexia whereas dried powder of stem with honey is taken orally to reduce pain and inflammation. The stem and leaf of the Cissus quadrangularis are used in food preparation in India. Numerous studies demonstrated that methanolic and petroleum ether extract of the whole plant are used in treating osteoporosis and fracture (Deka et al., 1994, Shirwaikar et al., 2003, Sharpa et al., 2007). It possesses antioxidant (Jainu and Shyamaladevi, 2005), anti-inflammatory, antibacterial (Kashikar and George, 2006, Thakur et al., 2009), antiviral (Balasubramaniana et al., 2010), and antiulcerogenic (Jainu and Shyamaladevi, 2004, Jainu and Shyamaladevi, 2006, Jainu et al., 2010) properties. It has also solved the problems of weight loss and various metabolic syndromes (Oben et al., 2006, Oben et al., 2007). Phytochemical studies of Cissus quadrangularis have shown the presence of several phytochemical constituents, such as ascorbic acid, flavonoids, and triterpenoids (Potu et al., 2010).
Inflammatory and pathogenic condition activates the enzymes cycloxygenase (COX) and 5-lipoxygenase (5-LOX). COX and 5-LOX are the key enzymes in the synthesis of prostanoid and eicosonoids from poly unsaturated fatty acids (PUFAs), which are involved in various inflammatory and allergic disorders (Martel-Pelletier et al., 2003, Daniel et al., 2004). Nuclear factor E2 p45-related factor 2 (Nrf-2) is a member of leucine zipper transcription family responsible for upregulation of genes of phase-II enzymes in response to oxidative stress and LPS treatment (Annukka et al., 2008). Heme oxygenase-1 (HO-1) is the phase-II enzyme catalyzes the degradation reaction of heme to convert biliverdin further to bilirubin. Deficiency of HO-1 isozyme reveals severe damage of iron metabolism, which leads to liver, and kidney oxidative damage and inflammation. Consequently HO-1 expression increases cell resistance to oxidative injury (Daniel et al., 2004). Inducible nitric oxide synthase (iNOS) and tumour necrosis factor alpha (TNFα) are the proinflammatory mediators. The concentration of iNOS is elevated in injury and inflammation, while TNFα is mainly formed through monocytes and activated macrophages. It is pleotropic proinflammatory cytokine with many actions like fever, macrophage activation, etc., that are central to the pathogenesis.
In the present study, we have examined the in vitro anti-inflammatory activity of different extracts of Cissus quadrangularis on COX, 5-LOX and proinflammatory mediators. Inhibition of COX-1 and COX-2 were evaluated by using chromogenic assay and inhibition of 5-LOX was evaluated by using polarigraphic method. Whole plant of Cissus quadrangularis was extracted with different organic solvents and the acetone extract gave the potential inhibition of COX and 5-LOX. Consequently number of partially purified fraction of acetone extract we have screened to examine the inhibition of COX and 5-LOX. Single active acetone fraction of Cissus quadrangularis have shown potent inhibition hence called AFCQ. AFCQ was further used to evaluate the eventual mechanism essentially its anti-inflammatory effect in LPS induced inflammation. We found that AFCQ have potentially inhibited COX-2, iNOS, and TNFα expression in LPS treated murine macrophages RAW 264.7 cells. Upregulation of HO-1 expression connected with activation and expression of Nrf-2 might be related in the anti-inflammatory activity of AFCQ.
Section snippets
Plant material
Plant material was collected from Vidarbh region of Maharashtra in March 2008. The plants were authenticated at the Department of Botany, Rashtrasant Tukdoji Maharaj Nagpur University, Nagpur and voucher specimens were deposited in the herbarium. Botanical name is Cissus quadrangularis and voucher specimen number is 9433.
Preparation of extracts of Cissus quadrangularis
The stem of Cissus quadrangularis was shade dried for five to six months, powdered, and extracted successively using soxhlet apparatus with hexane, chloroform, acetone, ethyl
Effect of different extracts on cycloxygenase enzyme activity.
Preliminary studies with the plant extracts of Cissus quadrangularis exhibited inhibitory activity against both cycloxygenase (COX-1 and COX-2) enzymes. The IC50 values of all extracts are obtained on the inhibitory effect on COX-1 and COX-2 enzymes were given in Table 1. Hexane, acetone, ethyl acetate, methanol, and ethanol extracts of Cissus quadrangularis showed satisfactorily inhibition of enzyme COX-1 and COX-2 in dose dependent manner (1000 μg/ml, 100 μg/ml, 10 μg/ml) in (Fig. 1a and b),
Discussion
Almost all diseases are characterized by the inflammation and pain in response to different conditions like injuries, infection, etc. Inflammation and pain indicate the presence of COX and 5-LOX. The enzyme COX has two isoforms, COX-1 and COX-2. COX-1 is the constitutive enzyme, COX-2 is an inducible enzyme and its level is always found elevated in abnormal conditions like inflammation, arthritis and in carcinogenesis. Non steroidal anti-inflammatory drugs (NSAIDs) inhibit COX-2, but have many
Conclusion
According to experimental results, AFCQ obtained from Cissus quadrangularis, was found to have better anti-inflammatory activity. AFCQ might be helping in the dissociation the Nrf-2-Keap-1 complex which in turn leads to activation of Nrf-2 protein by phosphorylation and translocation from cytoplasm into nucleus. Phosphorylated Nrf-2 binds with ARE which in turn activates the gene expression of phase-II enzymes. The antioxidant proteins like HO-1 get expressed through the Nrf-2 Pathway. It leads
Acknowledgements
We are express our gratitude to Mr. G. Durgaprasad, Mr. Rambabu, Mr. Kishore, Mr. Tanmay, Ms. Monima, Mr. Bharat, urmi, Dr. Anil Kotha, Dr. Chandramohan, Dr. Pulla Reddy and Dr. G.V. Reddy for their support and guidance. We are also grateful to UGC Networking Resource Center, School of Chemistry, University of Hyderabad for providing the laboratory, instruments and other facilities.
References (28)
- et al.
Mechanism for irreversible self deactivation of prostaglandin synthetase
Journal of Biological Chemistry
(1976) - et al.
Purification of the cyclooxygenase that forms prostaglandins: demonstration of two forms of iron in the holoenzyme
Journal of Biological Chemistry
(1976) - et al.
Gastroprotective action of Cissus quadrangularis extract against NSAID induced gastric ulcer: role of proinflammatory cytokines and oxidative damage
Chemico-Biological Interactions
(2006) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays
Journal of Immunological Methods
(1983)- et al.
Immunochemical evidence for the involvement of prostaglandin H synthase in hydroperoxide-dependent oxidations by ram seminal vesicle microsomes
Journal of Biological Chemistry
(1983) - et al.
Purification of arachidonate 5-lipoxygenase from potato tubers
Methods in Enzymology
(1990) - et al.
Selective inhibition of cyclooxygenase-2 by c-phycocyanin, a biliprotein from spirulina platensis
Biochemical and Biophysical Research Communication
(2000) - et al.
Anti-osteoporotic effect of ethanol extract of Cissus quadrangularis linn on ovariectomized rat
Journal of Ethnopharmacology
(2003) - et al.
The effect of phytogenic anabolic steroid in the acceleration of fracture repairs
Life Sciences
(1965) - et al.
Levonen enterolactone induces heme oxygenase-1 expression through nuclear factor-E2-related factor 2 activation in endothelial cells
The Journal of Nutrition
(2008)
Antiviral activity of ancient system of ayurvedic medicinal plant Cissus quadrangularis linn. (Vitaceae)
Journal of Basic Clinical Pharmacy
Mechanism of selective inhibition of the inducible isoform of prostaglandin G/H synthase
Proceedings of the National Academy Science of the United States of America
Effect of Cissus quadrangularis in accelerating healing process of experimentally fractured radius-ulna of dog: a preliminary study
Indian Journal of Pharmacology
Regulation of heme oxygenase-1 expression through the phosphatidylinositol 3-kinase/akt pathway and the nrf-2 transcription factor in response to the antioxidant phytochemical carnosol
Journal of Biological Chemistry
Cited by (40)
Chemical composition, cytotoxic and pro-inflammatory enzyme inhibitory properties of Withania somnifera (L.) Dunal root extracts
2022, South African Journal of BotanyCissus quadrangularis L: A comprehensive multidisciplinary review
2021, Journal of EthnopharmacologyImmunostimulatory and anti-allergic potential of novel heterotrimeric lectin from seeds of Zizyphus mauritiana Lam
2021, International Journal of Biological MacromoleculesEnhanced osteogenesis of 3D printed β-TCP scaffolds with Cissus Quadrangularis extract-loaded polydopamine coatings
2020, Journal of the Mechanical Behavior of Biomedical MaterialsCitation Excerpt :In contrast to this, the total amount of CQE released in pH 7.4 was 12.5, 14, and 15% for TCP, PD2, and PD10, respectively. The burst release behavior in acidic conditions can be beneficial as CQE and the ketosteroids it contains have shown possible anti-inflammatory properties (Bhujade et al., 2012), and mitigating the initial inflammatory response can be beneficial in accelerating the healing process (Yu et al., 2016). Following the cytotoxicity and drug release data, 200 and 500 μg were chosen as low and high CQE loading amounts on PD2 coatings for in vitro studies, respectively.
HRLC-ESI-MS based identification of active small molecules from Cissus quadrangularis and likelihood of their action towards the primary targets of osteoarthritis
2020, Journal of Molecular StructureCitation Excerpt :The stem of CQ has been consumed as both dietary ingredient and medicine by the local populations in India. Some in vitro and in vivo studies have addressed their effective actions to treat inflammation [5], rheumatoid arthritis [6], osteoporosis [7], diabetes [8], gastrointestinal problem [9] and bacterial infection [10], etc. Based on conventional literature, CQ stem has interesting pharmacological properties to cure all kind of bone-related illness and it has been a primary option to heal broken bones.