The Potential Protective Effect of Spirulina Nanoparticles Against Ehrlich Solid Tumor Bearing Mice Induced Liver Toxicity, Tumor Markers, DNA Fragmentation, Oxidative Stress and Monooxygenase Variations

Ehrlich solid tumor (EST) is a spontaneous murine mammary adenocarcinoma, undifferentiated, making mainly benecial for tumors studies. The current research aim to describe the antineoplastic activity of spirulina nanoparticles extract (SP NPs) against EST induced alteration in liver functions, oxidative stress, tumor markers, monooxygenase and DNA damage in female mice. 40 female mice were arbitrarily doled out into 4 gatherings (1st, Control; 2nd, SP NPs; 3rd, EST; 4th, EST + SP NPs). Results revealed signicant elevation in the levels of AST, ALT, GGT, ALP, AFP, CEA, DNA damage; thiobarbituric acid reactive substances (TBARS), and glutathione peroxidase (GPx) in liver homogenate and depletions in the levels of albumin, total proteins, cytochromes (CYP450 & cyt b5), reduced glutathione (GSH), glutathione reductase (GR), glutathione S transferase (GST) and superoxide dismutase (SOD) activity in liver homogenate compared to control. EST Treatment with SP NPs (EST + SP NPs) improved and modulates these variations in liver functions, tumor markers, DNA damage, cytochromes, enzymatic and non-enzymatic, and oxidative stress as compared to mice bearing EST. These results suggest that SP NPs may have hepatic protective, antioxidant, and anti-cancer properties; in addition to inhibition of the EST development, and liver DNA damage induced by EST.


Introduction
Cancer is the most genuine motivation of death around the world, in 2016, 1. Most methodologies treatments that utilized in disease therapy is chemotherapy that murder malignancy cells by instigating apoptosis and causes seriously in uence the existence of patients and address an immediate reason for death (Tousson et al., , 2016. Over 40% of remedial medications were delivered dependent on normal items and their subordinates in the previous many years . Spirulina is lamentous blue-green microalgae having a place with 2 distinct genera Spirulina and Arthrospira that comprises of around 15 species (Siva Kiran et al., 2015). In numerous nations of Africa, Spirulina used as food that full of protein and is accumulated from basic water, dried and eaten (Vedi et al., 2013). 70% of spirulina is protein in addition to carotenoid, omega 3 & 6, glycolipids, polysaccharides, sulfolipids, Gamma Linolenic Acid (GLA), provitamins; and many minerals as potassium, calcium, magnesium, iron, manganese, selenium and zinc (Khan et al., 2005). Pre-clinical and clinical assessments recommend that spirulina has certain helpful impacts as in decline in blood cholesterol, weight, radiation insurance, and nephrotoxicity (Deng and Chow, 2010;Siva Kiran et al., 2015).
Nanotechnology has an immense scope of utilization in conclusion, drug conveyance, food industry, ecological cleanup, paints, hardware, sportsbeauty care products, and sunscreens (Nel et al., 2006;El-Masry et al., 2020b). Nanoparticles (NPs) are materials with a size scope of roughly 1-100 nm . The little size of these particles gives an expanded surface region and incites interesting and explicit physicochemical qualities like high conductivity, and compound reactivity contrasted with mass materials

Ehrlich ascites carcinoma
Ehrlich ascites carcinoma (EAC) bearing mice were gotten from Egyptian National Cancer Institute in Cairo University. Gp4: (EST + SP NPs); mice were infused hypodermically with about 3,000,000 cells of EAC per mouse to start tumor (EST) and left for about fourteen days till the improvement of tumor at that point treated with spirulina nanoparticles separate (SP NPs) for an additional fourteen days.

Sample collection
Before the nish of the analysis, mice were euthanized with intraperitoneal infusion with sodium pentobarbital and afterward went through complete necropsy. Blood tests from each mouse were gotten from the vena cava and accumulated in non-heparinised glass tubes prior to being left for 30 minutes to clump at 37C o before their being liable to 5000 xg outward for ten minutes.
Sera were isolated and put away in aliquots at -80° C until required. Notwithstanding liver of each mouse was taken out, squashed and homogenized (10%, w/v) and the supernatant was gathered and put away at -80°C for the assurance of DNA harm, oxidative pressure, microsomes and cytochromes.   (Table 1). Conversely; a signi cant decline in the albumin and total proteins was estimated in EST when contrasted with control (Table 1). On the other hand, a signi cant decline was estimated in ALT, AST, ALP, and GGT in treated EST with spirulina nanoparticles (EST+SP NPS) (Table 1). Also, a signi cant elevation in albumin and total proteins in treated EST with spirulina nanoparticles (EST+SP NPS) ( Table 1).

Impacts of spirulina NPs on tumor markers
A signi cant elevation were assessed in AFP and CEA levels in EST when compared with control ( Table   2). On the contrary; a signi cant decrease in AFP and CEA levels in treated EST with spirulina nanoparticles (EST+SP NPS) were detected ( Table 2). 3.3. Impacts in DNA breakages in liver tissue 2.8. Mixed-function monooxygenase systems in liver microsomes The total microsomal hepatic substance of the cytochromes (cyt b5 and CYP450) was assessed by utilizing the strategy for Omura and Sato (1964). Amidopyrine N demethylase and aniline 4 hydroxylase activities were estimated by Nash (1953) and Kato and Gillette (1965), separately.
2.9. Liver homogenate biochemical assays TBARS and GSH were estimated as depicted by Tappel and Zalkin (1959) and Pastore et al. (2003) respectively. Conversely; GST and GPx were measured by the strategy for Habig et al. (1974) and Paglia and Valentine (1967) respectively. In addition to SOD and GR were measured by the strategy depicted by

Statistical analysis
Mean and standard blunder esteems were resolved for every one of the boundaries and the outcomes were communicated as mean ± standard mistake. The information were investigated utilizing a single direction examination of difference (ANOVA) trailed by Duncan various correlation. P < 0.05 was statically huge as per Norušis (2006). %change = Mean value of treated -Mean value of control / Mean value of control * 100 A signi cant elevation in DNA breakages was assessed in liver tissues in EST group when contrasted with control (Table 3). In distinction, a signi cant decline in DNA breakages in liver tissues in treated EST with spirulina nanoparticles (EST+SP NPS) as compared to EST (Table 3).

Effects of spirulina NPs on drug metabolism
The cytochromes (CYP450 & cyt b5), amidopyrine N demethylase and aniline 4 hydroxylase in mice liver showed normal activity in control and SP NPs groups (Table 5). Ehrlich solid tumor induced depletion in cytochromes (CYP450 & cyt b5) and elevation in Aniline4-hydroxylase as compared to control.
Conversely; treatment of EST with spirulina nanoparticles (EST+SP NPS) induced elevation in cytochromes (CYP450 & cyt b5) and depletion in Aniline4-hydroxylase as compared to EST (Table 4).

Changes in oxidative stress in liver tissue homogenate
A signi cant elevation was distinguished in TBARS, and GPx, in liver homogenates in EST group when contrasted with control (Table 4). Conversely; a signi cant decline in TBARS, and GPx in liver in treated EST with spirulina nanoparticles (EST+SP NPS) ( Table 5). A signi cant decline in GSH, GR, GST and SOD activities in liver homogenate in EST group as compared to control group were detected (Tables 4). In contrast; a signi cant elevation in GR, GST and SOD in liver homogenate in treated EST with spirulina nanoparticles (EST+SP NPS) ( Table 5).

Discussion
Malignant growth is a primary general medical condition worldwide and is the subsequent driving reason for death. Ebb and ow research plan to portray the antineoplastic action of spirulina nanoparticles separate against Ehrlich solid tumor actuated modi cation in liver functions, oxidative stress, tumor markers, monooxygenase and DNA damage in female mice. Our outcomes uncovered that; EST induced elevations in AST, ALT, ALP, and GGT, and exhaustions in total protein and albumin suggesting liver dysfunction. Notwithstanding; the treatment of EST with SP NPs (EST + SP NPs) improved the liver Present examination exhibited that; the medicines of EST with SP NPs was powerful in controlling cell reinforcement chemical exercises by decrease the degrees of TBARS, GPx and a critical height in GSH, GR, GST, SOD exercises. Spirulina contains a signi cant compound SOD that is 1700 units/g of dry mass that acts in a roundabout way by hindering the pace of oxygen revolutionary producing responses (Belay, 2002). Besides, Karadeniz et al. (2009) tracked down that; Pre-treatment with Spirulina platenesis may assume a part in lessening the poisonous impact of cadmium and its cancer prevention agent properties appeared to intervene a particularly defensive impact, shown by the decrease of MDA and NO just as the rise in levels of GSH and SOD. This is likewise in accordance with Hassanen et al. (2015) who consider the biochemical impacts of spirulina platensis against oxidative pressure brought about by doxorubicin.

Conclusions And Recommendations
Spirulina nano-particles have ability to improve liver functions and structure, inhibit the proliferations, AFP, CEA, and modulate the oxidative stress, electrolytes, DNA damage in breast cancer female mice. Whenever a rmed in people's creatures, these outcomes could demonstrate that spirulina nano-particles, as cell reinforcement, could be utilized as an assistant preventive treatment with chemotherapy drugs.

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Data availability:
All data used in this study are included in this published article.