Intraperitoneal Lavage with Crocus sativus Prevents Postoperative-Induced Peritoneal Adhesion in a Rat Model: Evidence from Animal and Cellular Studies

Abstract

Postoperative peritoneal adhesions are considered the major complication following abdominal surgeries. The primary clinical complications of peritoneal adhesion are intestinal obstruction, infertility, pelvic pain, and postoperative mortality. In this study, regarding the anti-inflammatory and antioxidant activities of Crocus sativus, we aimed to evaluate the effects of Crocus sativus on the prevention of postsurgical-induced peritoneal adhesion. Male Wistar-Albino rats were used to investigate the preventive effects of C. sativus extract (0.5%, 0.25% and 0.125% w / v ) against postsurgical-induced peritoneal adhesion compared to pirfenidone (PFD, 7.5% w / v ). We also investigated the protective effects of PFD (100 μg/ml) and C. sativus extract (100, 200, and 400 μg/ml) in TGF-β1-induced fibrotic macrophage polarization. The levels of cell proliferation and oxidative, antioxidative, inflammatory and anti-inflammatory, fibrosis, and angiogenesis biomarkers were evaluated both in vivo and in vitro models. C. sativus extract ameliorates postoperational-induced peritoneal adhesion development by attenuating oxidative stress [malondialdehyde (MDA)]; inflammatory mediators [interleukin- (IL-) 6, tumour necrosis factor- (TNF-) α, and prostaglandin E2 (PGE2)]; fibrosis [transforming growth factor- (TGF-) β1, IL-4, and plasminogen activator inhibitor (PAI)]; and angiogenesis [vascular endothelial growth factor (VEGF)] markers, while propagating antioxidant [glutathione (GSH)], anti-inflammatory (IL-10), and fibrinolytic [tissue plasminogen activator (tPA)] markers and tPA/PAI ratio. In a cellular model, we revealed that the extract, without any toxicity, regulated the levels of cell proliferation and inflammatory (TNF-α), angiogenesis (VEGF), anti-inflammatory (IL-10), M1 [inducible nitric oxide synthase (iNOS)] and M2 [arginase-1 (Arg 1)] biomarkers, and iNOS/Arg-1 ratio towards antifibrotic M1 phenotype of macrophage, in a concentration-dependent manner. Taken together, the current study indicated that C. sativus reduces peritoneal adhesion formation by modulating the macrophage polarization from M2 towards M1 cells.

Acetyl-11-Keto-β-Boswellic Acid (AKBA) Prevents Lipopolysaccharide-Induced Inflammation and Cytotoxicity on H9C2 Cells

“…It is well established that LPS, as an exogenous ligand of TLR4, induces inflammatory processes involved in severe cardiac injury [ 34 ]. LPS-mediated activation of initiates NF- κ B dependent signalling pathways, resulting in the overproduction of various proinflammatory mediators, such as COX-2, iNOS, IL-1 β , IL-6, and TNF- α [ 6 , 24 , 26 – 28 , 35 , 36 ]. Excessive production of proinflammatory cytokines plays a vital role in developing many inflammatory disorders, including sepsis-induced myocardial dysfunction [ 37 ].…”

“…The ELISA assay evaluated the IL-1 β , IL-6, TNF- α , and PGE2 levels as inflammatory mediators, which were carried out in accordance with the manufacturers' protocol, IBL company [ 6 , 7 , 24 , 26 , 28 ], USA. In summary, 1.5 × 10 6 cells were cultured in a 6-well plate overnight, and they were treated according to the experimental grouping section.…”

“…A real-time PCR technique was performed through the SYBR Green procedure's employment to assess the possible impacts of different AKBA concentrations on the levels of TNF- α , IL-1 β , IL-6, and COX-2 as well as iNOS related mRNA. Rotor-Gene 6000 was involved as a real-time reaction detection system, and GAPDH was considered a reference gene [ 7 , 24 , 26 ]. We procured the required real-time PCR primers in a similar design to those mentioned in previous studies [ 29 ]; the primer specificity was blasted and confirmed by applying NCBI Primer-BLAST.…”

“…The nitrite oxide metabolite levels were measured based on the measurement of nitrite (NO −2 ) as the stable and the final NO product by the Griess method described elsewhere [ 30 ]. The NO levels were assessed in the supernatant at 540 nm by using the standard curve of different nitrite concentrations [ 6 , 7 , 24 , 26 – 28 ].…”

“…The total thiol content was evaluated through a colourimetric method, which was set according to the reaction of total thiol content with Ellman reagent (5,5′-dithiobis (2-nitrobenzoic acid) (Sigma-Aldrich) [ 31 ]. This particular reaction results in the formation of yellow-coloured TNB (5-thio-2-nitrobenzoic acid) that can be quantified at 412 nm [ 6 , 7 , 24 , 26 – 28 ].…”