Thrombin-like serine protease, antiquorin from Euphorbia antiquorum latex induces platelet aggregation via PAR1-Akt/p38 signaling axis

Urs, Amog P.; Manjuprasanna, Vaddarahally N.; Rudresha, Gotravalli V.; Hiremath, Vilas; Sharanappa, P.; Rajesh, R.; Vishwanath, Bannikuppe S.

doi:10.1016/j.bbamcr.2020.118925

Cited by 6 publications

(7 citation statements)

References 49 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Thrombin/papain was used as a positive/protease control. Based on our previous studies, 25,40 the healing of excision wound takes 6–10 days and drupin (0.5 U) was applied twice a day to check its efficacy in wound healing. Hence, 0.5 U might be enough to enhance wound healing.…”

Section: Resultsmentioning

confidence: 99%

Drupin, a thrombin‐like protease prompts platelet activation and aggregation through protease‐activated receptors

Manjuprasanna

Urs

Rudresha

et al. 2021

J of Cellular Biochemistry

Self Cite

View full text Add to dashboard Cite

Hemostasis is a proteolytically regulated process that requires activation of platelets and the blood coagulation cascade upon vascular injury. Activated platelets create a thrombogenic environment and amplify the coagulation process. Plant latex proteases (PLPs) have been used as therapeutic components to treat various ailments by folk healers. One of the main applications of plant latices is to stop bleeding from minor injuries and to enhance wound healing activity. Although many studies have reported the pro‐coagulant activities of PLPs, an in‐depth investigation is required to understand the mechanism of action of PLPs on platelets. Here, the effect of PLPs on platelet aggregation was studied systematically to validate the observed pharmacological effect by folk healers. Among 29 latices from the Ficus genus tested, Ficus drupacea exhibited potent pro‐coagulant and thrombin‐like activity. Drupin, a thrombin‐like cysteine protease responsible for platelet aggregation was purified from F. drupacea latex. Drupin exhibits pro‐coagulant activity and reduces the bleeding time in mice tail. It induces platelet aggregation by activating mitogen‐activated protein kinases and the nuclear factor‐κB and PI3K/Akt signalling cascade, which, in turn, phosphorylats, cytosolic phospholipase A2 leading to the release of thromboxane A2 from the granules to activate the nearby platelets to aggregate. Furthermore, we investigated the involvement of protease‐activated receptors in drupin‐induced platelet aggregation using specific protease activated receptor 1 (PAR1) and PAR4 receptor antagonists. The results confirmed that the drupin‐induced platelet aggregation was mediated by both PAR1 and PAR4, synergistically. Overall, drupin reduces the bleeding time by exerting pro‐coagulant activity and induces platelet aggregation by activating the intracellular signalling cascade.

show abstract

Section: Resultsmentioning

confidence: 99%

Drupin, a thrombin‐like protease prompts platelet activation and aggregation through protease‐activated receptors

Manjuprasanna

Urs

Rudresha

et al. 2021

J of Cellular Biochemistry

Self Cite

View full text Add to dashboard Cite

show abstract

“…After incubation blots were washed with TBST and probed with HRP conjugated anti‐rabbit IgG (1: 10 000) secondary antibody for 2 h at room temperature. Further, blots were developed by enhanced chemiluminescence and visualized using Bio Imaging system (Alliance 2.1 UVITEC Cambridge image analyzer) 20,21 …”

Section: Methodsmentioning

confidence: 99%

“…Further, blots were developed by enhanced chemiluminescence and visualized using Bio Imaging system (Alliance 2.1 UVITEC Cambridge image analyzer). 20,21 2.11 | Quantitative real time polymerase chain reaction (qRT-PCR)…”

Section: Western Blot Analysismentioning

confidence: 99%

Inhibition of sPLA₂ enzyme activity by cell‐permeable antioxidant EUK‐8 and downregulation of p38, Akt, and p65 signals induced by sPLA₂ in inflammatory mouse paw edema model

Jayachandra

Gowda

Rudresha

et al. 2022

J of Cellular Biochemistry

Self Cite

View full text Add to dashboard Cite

The arachidonic acid (AA) metabolic pathway, plays a vital role in the production of eicosanoids by the action of pro-inflammatory secretory phospholipase A 2 (PLA 2 ). Release of eicosanoids is known to be involved in many inflammatory diseases. Identification of the inhibitory molecules of this AA pathway enzyme along with the regulation of intracellular signaling cascades may be a finer choice to develop as a powerful anti-inflammatory drug. In this regard, we have screened few cell-permeable antioxidant molecules Tempo, Mito-TEMPO, N,N'-Bis (salicylideneamino)ethane-manganese(II) (EUK)-134, and EUK-8 against proinflammatory sPLA 2 s. Among these, we found EUK-8 is a potent inhibitor with its IC 50 value ranges 0.7-2.0 µM for sPLA 2 s isolated from different sources.Furthermore, docking studies confirm the strong binding of EUK-8 towards sPLA 2 . In vivo effect of EUK-8 was studied in HSF-sPLA 2 -induced edema in mouse paw model. In addition to neutralizing the edema, EUK-8 significantly reduces the phosphorylation level of inflammatory proteins such as p38 member of MAPK pathway, Akt, and p65 along with the suppression of pro-inflammatory cytokine (interleukin-6) and chemokine (CXCL1) in edematous tissue. This shows that EUK-8 not only inhibits the sPLA 2 activity, it also plays an important role in the regulation of sPLA 2 -induced cell signaling cascades. Apart from the sPLA 2 inhibition, we also examine the regulatory actions of EUK-8 with other downstream enzymes of AA pathway such as 5-LOX assay in human polymorphonuclear leukocytes (PMNs) and COX-2 expression in carrageenan-λ induced paw edema. Here EUK-8 significantly inhibits 5-LOX enzyme activity and downregulates COX-2 expression. These data indicate that EUK-8 found to

show abstract

“…Approximately 130 different types of triterpenoids have been isolated so far from the various species of Euphorbia. According to reports, tertracyclic triterpenoids are one of the major triterpenoids in the Euphorbia species and were divided into four classes: cycloartanes , lanostane (85)(86)(87)(88)(89)(90)(91)(92)(93)(94), euphane (20)(21)(22)(23)(24)(25)(26)(27)(28)(29)(30)(31)(32)(33)(34)(35)(36)(37)(38)(39), and tirucallane (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19). Specifically, 9,19-cyclolanosta...…”

Section: Terpenesmentioning

confidence: 99%

Euphorbia neriifolia (Indian Spurge Tree): A Plant of Multiple Biological and Pharmacological Activities

et al. 2023

View full text Add to dashboard Cite

Although India has a well-established and growing economy surrounding synthetic drug chemistry with an antibiotic base, a large part of the population, especially in forested villages and tribal belts, is relying solely on plant-derived drugs. This is due to a lower number of side effects, low chances of resistance development against pathogenic microorganisms, as well as the diversity and affordability of such drugs. In the Indian subcontinents, Euphorbia neriifolia Linn. (EN) is one of the valuable plants from the big family of Euphorbiaceae, which is usually found in rocky and hilly areas. E. neriifolia was found to be useful in curing tumors, abdominal swelling, bronchial infection, hydrophobia, earache, cough and cold, asthma, leprosy, gonorrhea, spleen enlargement, leucoderma, snake bites, scorpion stings, and causing appetite improvement, etc. Different in vitro and in vivo experimental studies were performed to determine the antioxidant, anti-diabetic, immunomodulatory, anti-inflammatory, anti-arthritic, wound healing, anti-atherosclerosis, radioprotective, anti-anxiety, anti-convulsant, anti-psychotic, anti-thrombotic, dermal irritation, hemolytic, analgesic, anti-fertility, diuretic, anti-microbial, anti-diarrheal, and anti-carcinogenic activities of the various parts of EN. Several bioactive compounds, such as euphol, nerifoliol, taraxerol, euphonerins A–G, lectin, etc., were isolated from E. neriifolia and need to be investigated further for various biological activities (cardiovascular and neuronal diseases). In the pharmaceutical sector, E. neriifolia was selected for the development of new drugs due to its broad pharmacological activities. Therefore, in the present review, distribution, classification, morphological and microscopical description, phytochemical investigation, pharmacological activities, medicinal uses, harmful effects, and their treatment were evaluated, especially against different lifestyle-related diseases.

show abstract

Thrombin-like serine protease, antiquorin from Euphorbia antiquorum latex induces platelet aggregation via PAR1-Akt/p38 signaling axis

Cited by 6 publications

References 49 publications

Drupin, a thrombin‐like protease prompts platelet activation and aggregation through protease‐activated receptors

Drupin, a thrombin‐like protease prompts platelet activation and aggregation through protease‐activated receptors

Inhibition of sPLA₂ enzyme activity by cell‐permeable antioxidant EUK‐8 and downregulation of p38, Akt, and p65 signals induced by sPLA₂ in inflammatory mouse paw edema model

Euphorbia neriifolia (Indian Spurge Tree): A Plant of Multiple Biological and Pharmacological Activities

Contact Info

Product

Resources

About

Thrombin-like serine protease, antiquorin from Euphorbia antiquorum latex induces platelet aggregation via PAR1-Akt/p38 signaling axis

Cited by 6 publications

References 49 publications

Drupin, a thrombin‐like protease prompts platelet activation and aggregation through protease‐activated receptors

Drupin, a thrombin‐like protease prompts platelet activation and aggregation through protease‐activated receptors

Inhibition of sPLA2 enzyme activity by cell‐permeable antioxidant EUK‐8 and downregulation of p38, Akt, and p65 signals induced by sPLA2 in inflammatory mouse paw edema model

Euphorbia neriifolia (Indian Spurge Tree): A Plant of Multiple Biological and Pharmacological Activities

Contact Info

Product

Resources

About

Inhibition of sPLA₂ enzyme activity by cell‐permeable antioxidant EUK‐8 and downregulation of p38, Akt, and p65 signals induced by sPLA₂ in inflammatory mouse paw edema model