Quantitative analysis and antioxidant activity
The yield obtained after extraction is 5.03%, this result is lower than recorded by
Mahmoudi et al., (2016) who obtain 9.8%. This difference in yields can be influenced by several factors such as chemical composition, physical characteristics of the plant material, extraction method and other conditions
(Dai and Mumper, 2010).
The results of the total polyphenols of the Ethanolic extract show that the
F.
carica extract has a polyphenol content of 42.14±0.14 µg E AG/mgE. This result is close to those obtained by
(El-Shobaki et al., 2010) while
Mahmoudi et al., (2016) found that the total polyphenol contents of leaves extracts of ten varieties of
F.
carica in Algeria ranged from 52.296±5.232 - 48.973± 2.015 (µg E.AG /mg E). The content of flavonoids are quantified 30.85±2.85 µg E.Q/mg E. This result is similar to other data obtained by
Lahmadi et al., (2019) who found 33.52±1.34 and higher than those obtained by
Mahmoudi et al., (2016) who found 11.29; µg E.Q/mg E and 14.388 µg E.Q/mg E respectively. These observed differences in polyphenol and flavonoid contents can be attributed to several factors such as the used extraction method, the degree of maturity of the plant.
The antioxidant capacity of the crude extract determined from IC50 is estimated to be 420 µg/ml, this result is less than that obtained by
Lahouel et al., (2016) who found a value of 275.23 µg/ml. Fig leaves contain a Considerable number of beneficial compounds namely Polyphenols and Flavonoids, which act as antioxidants
(El-Shobaki et al., 2010). This antioxidant capacity of fig leaves is significantly correlated with Phenolic content
(Mahmoudi et al., 2016).
The weight gain and relative weight growth
The results of the weight gain evaluation show a significant decrease (P≤0.001) between the control group (G1) and the Deltamethrin-treated group (G4) with a non-significant increase (p>0.05) between G4 and groups 5 and 6. (Fig 1).
Fig 2 and 3 show the results of the relative weight examination of the liver and kidneys., the findings indicate a significant decrease (p≤0.001) in the relative weight of the liver between G4 and G1 and a slight increase in the relative weight is recorded in the groups G5 and G6.
On the other hand, data show a non-significant decrease in the relative weight of kidneys in G4 compared to the control and a non-significant increase in the group G5, G6 compared with G4. The weight loss is most likely due to a reduction in food intake and the harmful effects of Deltamethrin. Our results are similar to those found by
Saoudi et al., (2011) who worked on different pyrethroid compounds (Deltamethrin, Fenvalerate and Diazinon), indicates a non-significant drop in relative weight in the DLM treated group as compared to the control group. This decline might be attributed to the impact of toxins on key organs including as the liver, kidney and rat
(Kara et al., 2005).
The slowing of body growth in treated rats can be explained by the disruption of cellular metabolism under the effect of oxidative stress generated by “reactive oxygen species” (ROS) witch The most significant endogenous sources of DNA damage are capable of inducing numerous modifications, including lesions to nucleotide bases, as well as effects on lipid oxidation, which represents the utilization of energy substrate. Lipid peroxidation, the degradation of membrane fatty acids and protein damage are also among the notable consequences and other chemical mediators such as a few pro-inflammatory cytokines which the body may discharge upon contact with toxicants like pesticides
(Lohiya et al., 2017).
Biochemical parameters
The obtained results of Table 1 show that the group treated with deltamethrin presents a significant increase (P≤0.001) in the biochemical parameters (Glucose, Triglyceride, Cholesterol) as well as a significant increase of the activity of TGO and no significant change of the TGP, the same results are recorded for the indicators of renal activity (creatinine, urea) compared to the control group.
In addition, no significant change was observed in total protein, only a slight increase. Data show that the extract of the leaves of
F.
carica is able to decrease the toxicity induced by the DLM, where we recorded a significant decrease in the rate of the biochemical parameters of G5 and G6 compared with G4.
The comparison by the Tuky test showed a no-significant difference between the groups treated by the combination and the control group except for the cholesterol (a significant difference and the decrease is very slight). According to our findings, rats treated with Deltamethrin has significantly higher glucose levels than rats of the control group, these findings are consistent with the study of
Manna et al., (2005)., this increase of glucose level is due to the entrainment of toxic substances of emotional reactions in the limbic system, which induce an rise in glucagon secretion while also causing an insulin decline. As a result, glucose is produced from hepatic glycogen and enters the bloodstream
(Eraslan et al., 2007).
A normal level of plasma glucose is observed in rats treated with
F.
carica, These findings are consistent with the work of (
Stephen Irudayaraj et al., 2017), possibly as a result of extract’s potentiation of pancreatic insulin production from regenerated β-cells
(Sunil 2012).
The group receiving treatment with
F.
carica and Deltamethirin exhibited improved blood glucose levels., this improvement can be explained by
F.
carica extract stimulates glucose absorption in rat peripheral tissues in a dose-dependent manner
(Stephen Irudayaraj et al., 2017). F.
carica improve the insulin sensitivity
(El Hilaly and Lyoussi, 2002).
Triglycerides in treated rats are higher than in controls, indicating that Deltamethirin causes hepatic hypertri glyceridemia. These results align with previous researchers’ findings and which can be explained by the excess of hepatic triglyceride synthesis or by the reduction of triglyceride hydrolysis caused by inhibition of the lipolytic enzyme
(Eraslan et al., 2007). Rats given the plant exhibit a significant reduction in TG levels compared to controls. This can be explained by production of triglyceride precursors such as acetyl-CoA and glycerol phosphate
(Eddouks et al., 2005). Our results are similar to the results of
Joerin et al., (2014), who confirmed the anti-hyperlipidemic effect of
F.
carica leaf extract.
Cholesterol level in MLD-treated rats are significantly higher than in control. Our results are consistent with the study of
Golli-Bennour et al., (2019), This increase can be attributed to the effect of pesticides on hepatic cell membrane permeability and liver dysfunction
(Yousef et al., 2006), which is confirmed by the increase of TGO, TGP and glucose.
The same results are recorded in the groups treated with the plant compared to the control, so the plant doesn’t show an improvement and this may be due to the short duration of co-treatment with the extract, these results corroborate with the study of
Joerin et al., (2014). The increase in GOT and GPT levels in the MLD treated group compared to the control groups is similar to that recorded by
(Manna et al., 2005).
Although Pyrethroids have a selective mechanism of action, their harmful side effects also include severe liver dysfunction, interruption of the production of these enzymes and altered liver membrane permeability
(Saoudi et al., 2011). The co-treatment with
F.
carica show a decrease of GOT and GPT of G2 and G3 compared to the control group, our results are similar to those of
Fouad et al., (2019) who reported that flavonoid extract of
F.
carica has marked scavenging activities against hydroxyl and super oxide anion free radicals. A possible mechanism of
F.
carica extract as a hepatoprotector may be due to its antioxidant effect or inhibition of cytochrome P450s.
Preliminary phytochemical studies indicate that the Methanolic extract of
F.
carica leaves contains steroids, triterpenoids and their glycosides and coumarins. According to
Oh et al., (2002), coumarins (phenolic compounds) exhibit hepatoprotective action, hence it is possible that the contents of
F.
carica are responsible for its demonstrated protective effect.
Our study shows a slight increase in total protein levels after exposure of rats to MLD compared to control rats. These results are confirmed by the study of
Saoudi et al., (2011). Indeed, when environmental stresses (water stress, temperature, oxidative stress, exposure to pollution and infection by pathogens...) are severe, most proteins are denatured
(Mohammad and Heidari, 2008).
The study demonstrated a significant increase in protein levels after using
F.
carica extract, indicating the presence of cytoprotective molecules such as phenolic compounds that improve the homeostasis of the biochemical parameters. This is attributed to the antioxidant properties of polyphenols containing the catechol group, as reported by
Brglez Mojzer et al. (2016). The study also observed an increase in urea and creatinine levels in DLM-treated groups compared to the control group, indicating the harmful effects of DLM on nephron excretory function. This is consistent with previous research that showed similar results with other pesticides. However, the group treated with
F.
carica extract showed a significant decrease in creatinine and urea levels, indicating its antioxidant properties mainly due to flavonoids and phenolic compounds. These findings are consistent with the findings of
Kore et al., (2011) who showed that
F.
carica extracts significantly reduce creatinine and nephrotoxicity caused by gentamicin. The results indicate the beneficial role of F. carica on renal function and its ability to reduce the harmful effects of DLM.
Histopathological results
The observation of the histological sections of G1 show a normal cellular architecture of the liver with normal hepatocytes. On the other hand, the sections obtained of G4 show an inflammatory infiltrate; hepatocyte cells take a ballooning aspect, acute hepatitis, loss of trabecular architecture (intra lobular necrosis) and clarification of hepatocytes cytoplasm. Group 5 show the presence of an inflammatory infiltrate with little vascularity, clarification of the hepatocytes and vascular congestion (dilation of the vessels). On the other hand, group 6 show a clear improvement with a small vascular congestion (Fig 4).
Our findings are consistent with previous research on DLM. By
Chargui et al., (2012). Deltamethrin exposure induced apoptosis; by increasing mRNA expression of apoptotic markers Bax and caspase-3 as well as decreasing bcl-2 in liver tissues
(Maalej et al., 2017).
On the other hand, liver tissue apoptosis is clearly reversed by co-treatment with
F.
carica. In particular, it is shown that the extract dose of 400 mg/kg is more efficient to reduce apoptosis. This improvement is a result of the flavonoid present in
F.
carica leaves
(Ammar et al., 2015).
Furthermore, the DLM exposure increase the expression of inflammatory markers in primary rat hepatocytes. Hepatic slices with mild vasoconstriction can be explained by the reduction response of the liver (pressure is elevated). As a result of its toxic action, DLM caused acute hepatitis and liver damage. However, the recombination of DLM with
F.
carica extract indicates normal hepatocytes due to the plant’s hepatoprotective function. A proposed mechanism of Ficus extract. Ethanolic extracts of
F.
carica. leaves at 600 mg/Kg exert a high anti-inflammatory effect of 75.90% in acute inflammation and 71.66% in chronic inflammation by reducing granuloma weight
(Patil and Patil, 2011).
Histological sections (Fig 5) show that the kidneys in the control group have a normal architecture (nephrons have glomeruli, glomerular capsules, Bouman capsules and renal tubules in the cortical and medullary areas), without any inflammatory changes.
On the other hand, several histological changes can be seen in G4, including an inflammatory filtrum of a lymphoplasmacytic (interstitial nephritis between the nephrons), slight vascular congestion and vessel enlargement (due to increased pressure in the kidneys). However, the group 5 figure shows a slight atrophy of the fibrillation renal tubules and group 6 figure show, a small improvement with vascular congestion, a sign of interstitial nephritis around the glomeruli (lymphoplasmacytic infiltrate). These results support the biochemical data. Our findings are consistent with those of
Poonam et al., (2014), which can be explained by the short-term toxic effects of Deltamethirine on the kidneys, however, in the groups that received
F.
carica extract, a slight atrophy of the fibrillated renal tubules is observed, a sign of interstitial nephritis around the glomeruli. These findings support those of
Kore et al., (2011), which demonstrate that the plant reduces the impact of pesticide, due to the brief course of therapy, the kidneys are not entirely healthy. However, treatment of
F.
carica extract significantly reduced the inflammation., this can be explained by the
in vivo analysis that detected a notable anti-inflammatory regulatory effect of
F.
carica (Patil and Patil, 2011).
