Qualitative Analysis of Phytochemicals:
Phytochemical findings in (Table 1) suggest the presence of phytochemicals in R.officinalis leaves such as steroids, saponins, tannins, anthocyanins, emodins, flavonoids, terpenoids, glycosides and alkaloids and phenols in two extracts. In contrast, fatty acids were absent in ethanol and methanol extracts, whereas emodins and alkaloids were absent in methanol extract.
This data agree with Edrah et al. (2017), who reveal that the R.officinalis ethanolic extract contains flavonoids, terpenoids, glycosides, and phenols, whereas saponins have been absent. Also, in the same line with Khamis and Aly (2017), who found that terpenoids, alkaloids, glycosides, saponins, and flavonoids were widely present in ethanolic extract R.officinalis. These plants were used as preservatives for food and as folk medicines. Terpenoids are used in cough, asthma, and hay fever treatments. Saponins have antibiotic properties and guard against hypercholesterolemia (Mir et al. 2013).
Table 1
phytochemical Screening tests of R.officinalis methanol and ethanol extracts
Chemical
Constituents
|
R.officinalis
|
Ethanol
80%
|
Methanol
80%
|
Steroids
|
++
|
+
|
Terpenoids
|
++
|
+
|
Tannins
|
++
|
++
|
Saponins
|
+++
|
++
|
Anthocyanins
|
+++
|
+
|
Emodins
|
+
|
ـ
|
Alkaloids
|
+
|
ـ
|
Glycosides
|
++
|
+
|
Flavonoids
|
++
|
+++
|
Phenols
|
+++
|
++
|
Fatty acids
|
ـ
|
ـ
|
* (+++), (++), (+) and (-) refer to high, moderate, low and absent amount, respectively. |
Antioxidant Activity
The DPPH is a stable free radical at room temperature and accepts an electron/hydrogen radical to become a stable diamagnetic molecule (David et al. 2004). The DPPH is usually used as a substrate to evaluate the antioxidant vigor (Edamatsu et al. 1989). The antioxidants react with the stable free radical DPPH and turn into it to 1,1-diphenyl-2-picryl hydrazine with decoloration (Kumar et al. 2012). DPPH radical's reduction potential is certain by the dwindling in its 517 nm absorbance, which is caused by antioxidants. Visually visible, like a change in color from purple to yellow. The latest findings for R.officinalis extract manifest free radical scavenging vigor (Table 3). The highest DPPH scavenging vigor is manifest by ethanol (80%) with R.officinalis extract.
Results manifest that aqueous methanolic extract (80%) extract of rosemary exhibited inhibition of 50 % while ethanolic extract 53.8%. Therefore the aqueous ethanolic extract (80%) is the more efficient solvent for extracting the phytochemical compounds from R.officinalis. This extract may contain plentiful phenolic and flavonoid compounds identified as nutritional and medicinal properties (Shanmugavel et al. 2018). in the same line with data gained by Gîrd et al. (2017), who found that the ethanol extract of R.officinalis contains a high amount of phenolic compounds which consider as antioxidant vigor of plant materials (Kim et al. 2011).
Table 3
Antioxidant Activity ofR.officinalis*
Solvent
|
%inhibition
|
IC50(µg/ml)
|
Rosemary
|
Rosemary
|
Methanol extract
|
50
|
62.87
|
Ethanol extract
|
53.8
|
54.5
|
*The IC50values correspond to the amount of extract required to scavenge 50% of radicals present in the reaction mixture.
|
Effect of R .officinalis extract on hematological parameters in blood rats exposed to HV 2 and 4 h.
In this study the hematological tests of the animals treated for 30 days with the ethanolic extract R.officinalis were carried out. Table (1) shows no presumed divergence in the scale of Hb, WBCs, and RBCs relative to the control. It was estimated that Hb (P < 0.05) decreased by around 15 and 11 % in the exposed HV groups for 2 or 4 h, compared with the control group, respectively. In the meantime, remedy groups with ethanolic R.officinalis extract increased probably (P < 0.05) this reduction of Hb and restored it to normal control. RBCs (P < 0.05) were assumed to decrease by about 12 and 6 % respectively in the exposed group to HV for 2 or 4 h compared to the control group. Meanwhile, preremdy groups with ethanolic R.officinalis extract improved presumed and returned RBCs to normal control. WBCs (P < 0.05) were assumed to increase by about 22 percent and 54 percent respectively in the exposed group to HV for 2 or 4 h compared to the control group. Packed cell volume (PCV) was thought to decrease by about 24 % and 30 % in the exposed group to HV for 2 or 4 h, respectively, compared to the control group. In the meantime, preremdy groups with ethanolic extract of R.officinalis strengthened presumed PCV and took it back to normal regulation. The RBCs are assumed to dwindle (P < 0.05) by around 12 % and 6 % relative to the control group in the exposed group to HV for 2 or 4 h, respectively. In the meantime, presumed preremdy groups with ethanolic extract of R.officinalis strengthened and returned RBCs to normal regulation. WBCs are assumed (P < 0.05) to increase by around 22 % and 54 %, respectively, in the exposed group to HV for 2 or 4 h relative to the control group. Meanwhile, it was thought that preremdy groups with ethanolic R.officinalis extract (P < 0.05) decreased this increase in WBCs by around 9 % compared to the 2 h exposed group to HV. In contrast, this refining was 25 % compared to the 4 h exposed group to HV.
Xiao-Feng and Gun confirm the current data (2017) 's data, who found the elevation of white blood cells in rats exposed to HVTL and radiation exposure (Hsu et al. 2010). Also, in harmony with about hematological parameters (Eid et al. 2015). The depletion in the values of hematological parameters following EMF radiation may be attributed to direct deterioration caused by radiation and overproduction of ROS by microwave radiation interaction, causing hemolysis (Aweda et al. 2004). The dwindling of RBCs may be due to the interaction between heme (iron) and SMF, where the magnetic field penetrates the body and acts ions in all organs, altering the cell membrane's potential disrupting the ions (Kula and Drozds 1996).
Table 4
Effect of R.officinalis (5 mg / kg b.w) extract hematological parameters in blood rats exposed to HV 2 and 4 h.
Groups
|
HB
(mg/dl)
|
PCV (mg/dl)
|
RBCs (106/mm3)
|
WBCs(103/mm3)
|
Control
|
13.88 ± 0.64
|
0.37 ± 0.05
|
6.34 ± 0.41
|
12.33 ± 1.66
|
R
|
14.85 ± 0.27
|
0.37 ± 0.03
|
6.52 ± 0.15
|
12.05 ± 1.62
|
HV2h
|
11.80a ± 0.89
|
0.28a ± 0.15
|
5.56a ± 0.69
|
15.00a ± 1.01
|
HV4h
|
12.40 ± 0.78
|
0.26a ± 0.06
|
5.94 ± 0.48
|
19.03a ± 6.27
|
HV2h&R
|
14.20 ± 1.39
|
0.35b ± 0.03
|
6.14 ± 1.11
|
13.70b ± 0.86
|
HV4h&R
|
14.50 ± 1.52
|
0.33b ± 0.05
|
6.55 ± 1.27
|
14.25ab ± 3.03
|
Data represent the mean ± S.E. of observations from eight rats. a Significantly different from control group at P < 0.05. b Significantly different from HV group at P < 0.05.
|
Effect of R.officinalis extract on liver function in serum of rats treated with HV:
These enzymes are essential to biological processes occurring within a living organism's body, and are one of health indicators. Results in Table (2) show that there was no presumed 30-day difference between R.officinalis extract remdy groups as compared with normal AST and ALT vigor regulation. Meanwhile, HV exposure for 2 or 4 h elevated presumed (P < 0.05) AST serum vigor is approximately 50 % and 61 % respectively, while ALT vigor is 30 % and 36 % compared with control group.
The size of these enzymes in serum, however, showed a suspected diminishing (P < 0.05) by handling either R.officinalis extract for 2 or 4 h at the same time as HV exposure. In contrast with groups exposed to HV for 2 or 4h, respectively, AST vigor was manifested to be less by 27 % and 31 %. R.officinalis extract simultaneously with the presumed sensitivity to HV (P < 0.05) reducing AST vigor by about 30 percent and 34 percent compared to HV for groups of 2 or 4 h, respectively. Also, data shows that there was no presumed divergence in the TB, DB and IB scale of 30 days between R.officinalis extract remedy groups. Meanwhile, the serum TB scale increased by about 42 % and 65 % (P < 0.05) for 2 or 4 h, while the DB scale increased by 97 % and 140 %, while the IB scale increased by 18 and 32 % relative to the control group. However, the TB serum scale indicated a presumed (P < 0.05) decrease in either R.officinalis extract handling simultaneously with 2,4h HV exposure. TB scale decreases by about 20 % and 37 % compared to 2, 4h, respectively. At the same time, the DB serum scale showed a presumed (P < 0.05) decrease in handling with R.officinalis extract in accordance with HV treatment of approximately 22 percent and 27 percent compared with HV groups for 2 and 4 hours, respectively.
Although IB serum scale was assumed to diminish (P < 0.05), handling with either R.officinalis extract was simultaneously with 2 or 4 h HV treatment and restored to almost control. With accordance to these data Salem et al. ( 2005) postulated that the damaging impactof static magnetic field on the liver is manifested by an raise in blood AST and the most specific marker of liver cell deterioration ALT vigor. The exposure to ELF-EMF of 2 mT and 50Hz raised presumed serum transaminases vigor in mice. Hashem and El-Sharkawy (2009). ALT and AST vigor improved presumed (P<0.05) for two months in serum and liver tissue homogeneous to EMF exposed rats in the same manner as standard control group Eid et al. (2015). These results coincide with the findings of abundant authors who found a presumed increase in the scale of enzyme vigor, ALT and AST in rats exposed to electromagnetic field radiated from base stations on cell phones with a frequency equal to 900 MHz (Sharma et al. 2017). Reduction of both transferases as a result of radiation released by base station due to liver parenchymal damage, suggesting liver dysfunction (Achudume et al. 2012).The vigor of all liver function tests raised as a result of raise in the intensity of magnetic field and mostly affected with a percentage > 90% (Ibrahim et al. 2008). Also, the results in the current work are consistent with the previous observation of Hashish et al. (2008) who reported that exposure to a 50-Hz ELF EMF for 30 days caused a presumed raise in AST scale which may be due to oxidative stress or apoptosis caused by the ELF EMF (Emre et al. 2011). EMFs induced structural changes in hepatocytes, primarily in mitochondria and induced liberation of free radicals and oxygen species which can induce liver disease which also the main organ of detoxification (Eid et al. 2015). Exposing to EMFs in humans or animals resulted in increasing glucocor_ticoids (cortisol) which enhance transamination processes, stress oxidative compounds, and produced hipoxy. This is an substantial reason for increasing the amount of transaminases in trialal groups. Hipoxy produc_tion could raise the AST and ALT value in serum, up to thousands of units in liter (Kulkarni and Gandhare 2015).Rosemary essential oils exhibited on refinment in the actions of ALT and AST enzymes in hypercholesterolemic rats (Hassanen 2015) or after lead exposure (Abd El Kader et al. 2012).These may be attributed to the stabilizing ability of the cell membrane preventing enzymes leakages (Pari and Karthikesan, 2007). These result in the same line with abundant authors who found that the serum bilirubin scale in trial rats was higher than both control group exposure to EMR (Sharma et al. 2017) and EMF exposure group(Berrahal et al. 2011). Along with the apparent presumed impacton bilirubin, a byproduct of red blood cell lysis prove the possibility of hemolysis. Previously (El-Bediwi et al. 2011) also reported a rise in the bilirubin in rats following cell phone exposure. Bilirubin values increased as a result of magnetic field strength (1.4 mT), and it was also influenced by magnetic field penetration to 50 Hz. Ibrahim et al. (2008) increased the amount of bilirubin serum after exposure relative to control. The elevation in the serum bilirubin scale can result from deteriorating cells leaking into circulation after magnetic field exposure (Novikov et al. 1999).
Table 5
Effect of R.officinalis (5 mg / kg b.w ) extract on liver functions in rats exposed to H V 2 and 4 h.
Groups
|
AST
(U/ml)
|
ALT
(U/ml)
|
T.Bilirubin (mg/dl)
|
D.Bilirubin
(mg/dl)
|
Ind.Bilirubin (mg/dl)
|
Control
|
40.10 ± 0.3
|
45.17 ± 0.52
|
1.02 ± 0.01
|
0.30 ± 0.06
|
0.73 ± 0.02
|
R
|
40.20 ± 0.04
|
43.06 ± 0.96
|
1.03 ± 0.03
|
0.26 ± 0.01
|
0.73 ± 0.05
|
HV2h
|
60.13a ± 0.05
|
58.84a ± 0.04
|
1.45a ± 0.05
|
0.59a ± 0.03
|
0.86a ± 0.02
|
HV4h
|
64.56a ± 0.03
|
61.44a ± 0.05
|
1.68a ± 0.06
|
0.72a ± 0.02
|
0.96a ± 0.04
|
HV2h&R
|
43.83ab ± 0.04
|
54.83ab ± 0.03
|
1.16ab ± 0.04
|
0.45ab ± 0.02
|
0.73b ± 0.01
|
HV4h&R
|
44.51ab ± 0.04
|
56.11ab ± 0.01
|
1.23ab ± 0.02
|
0.53ab ± 0.06
|
0.72b ± 0.07
|
Data represent the mean ± S.E. of observations from eight rats. a Significantly different from control group at P < 0.05. b Significantly different from HV group at P < 0.05.
|
Effect of R.officinalis extract on Total Protein and Albumin parameters in the serum of rats treated with HV
Results in Table (3) indicate no presumed difference in the 30-day serum protein and albumin scale between R.officinalis extract remedy groups compared with the control group. Meanwhile, HV exposure for 2 or 4 h increased the serum protein scale by around 39.5% and 43.1%, respectively (p > 0.05), while the albumin scale increased by 58% and 75% compared to the control group.
In a group remedy with R.officinalis, the serum protein scale was assumed to dwindle by around 20 % simultaneously with HV treatment for 2 or 4 h. In conjunction with exposure to HV for 2 or 4 h, R.officinalis also decreased albumin by around 17 % compared with HV for groups of 2 or 4 hours. These results are in agreement with those data reported by plentiful authors (Hashem and El-Sharkawy 2009). The increase in the concentration of serum protein and albumin were observed as a result of deteriorating cells that flow into circulation after exposure to the magnetic field (Ibrahim et al. 2008). Kulkarni and Gandhare (2015) found that total protein, albumin, and globulin display a transient 30-hour increase that decreases at 60 hours relative to the regulated community. The period of intensity and exposure of EFs has been found to play a vital role in inducing internal fields and activating biological differentiation.
Table 6
Effect of R.officinalis extract on serum total protein and albumin in rats exposed to HV 2 and 4 h.
Groups
|
Protein (mg/dl)
|
Albumin
(mg/dl)
|
Globulin (g/dl)
|
Albumin/globulin
|
Control
|
7.70 ± 0.03
|
4.12 ± 0.01
|
3.57
|
1.15
|
R
|
7.91 ± 0.04
|
4.31 ± 0.05
|
3.57
|
1.20
|
HV2h
|
10.74a ± 0.08
|
6.51a ± 0.04
|
4.28
|
1.51
|
HV4h
|
11.02a ± 0.02
|
7.21a ± 0.05
|
3.84
|
1.87
|
HV2h&R
|
8.60ab ± 0.06
|
5.41ab ± 0.06
|
3.23
|
1.66
|
HV4h&R
|
8.85ab ± 0.05
|
5.61ab ± 0.03
|
3.29
|
1.69
|
Data represent the mean ± S.E. of observations from eight rats.aSignificantly different from control group at P < 0.05.bSignificantly different from HV group at P < 0.05.
|
Effect of R.officinalis extract on kidney function parameters in the serum of rats treated with
HV:
Results in Table (4) show that there were no presumed divergences in urea, uric acid, and creatinine scale after 30 days between R.officinalis extract remedy groups relative to regular control.
Table 7
Effect of R.officinalis extract on kidney function in rats exposed to HV 2 and 4 h.
Groups
|
Urea
(mg/dl)
|
Creatinine
(mg/dl)
|
Uric Acid
(mg/dl)
|
Control
|
28.41 ± 0.02
|
2.64 ± 0.05
|
4.59 ± 0.02
|
R
|
28.47 ± 0.02
|
2.92 ± 0.03
|
4.63 ± 0.02
|
HV2h
|
35.31a ± 0.15
|
4.81a ± 0.02
|
7.98a ± 0.02
|
HV4h
|
39.63a ± 0.06
|
5.49a ± 0.03
|
8.33a ± 0.01
|
HV2h&R
|
33.37ab ± 0.05
|
3.29ab ± 0.02
|
6.29ab ± 0.02
|
HV4h&R
|
33.91ab ± 0.03
|
3.52ab ± 0.02
|
6.55ab ± 0.01
|
Data represent the mean ± S.E. of observations from eight rats.aSignificantly different from control group at P < 0.05.bSignificantly different from HV group at P < 0.05.
|
These findings are consistent with abundant authors who reported no presumed divergence in kidney function parameters in rats that feed on rosemary (Hassanen 2015). Rosemary aqueous extracts also contained high antioxidant content, such as certain phenolic compounds. They are rosemary and carnosic acids (El-sherif and Issa 2015). Meanwhile, HV treatment for 2 or 4h increased the serum urea scale by approximately 24 % and 40 %, while creatinine increased by around 82 % and 108 %, while uric acid scale increased by about 74 % and 82 % compared to control group. However, the urea, uric acid, and creatinine serum scale showed a suspected diminishing (p>0.05) in the handling of R.officinalis extract. R.officinalis, in combination with 2 h exposure to HV, indicates a small decrease in concentration of urea. The urea scale decreased by approximately 14 % after 4h relative to the group exposed to HV for 4h. On the other hand, creatinine was assumed to be reduced by about 32 %, and 36 % by R.officinalis extract simultaneously as HV exposure (p>0.05) compared with groups exposed to HV 2 or 4 h, respectively. Also, R.officinalis extracts reduced uric acid by around 22 % simultaneously with the assumed exposure to HV (p>0.05). The current findings are consistent with Sharma et al. (2017). In rats exposed to electromagnetic radiation, those who reported serum uric acid, urea, creatinine (p>0.05) were presumed to have increased. Irradiation can cause DNA molecules to split, and their bases (purines) broken, which can be catabolized into uric acid (Ganong 1999). Creatinine is predominantly produced in the muscles, and freely appears in the blood plasma and urine. Also, a reported rise in serum urea (p < 0.05) and creatinine due to sensitivity to extremely low-frequency electrical fields (Kulkarni and Gandhare 2015) and exposure to EMF of 2 mT, 50Hz (Hashem and El-Sharkawy 2009).
Effect of R.officinalis extract on oxidative stress in liver tissue homogenate of rats treated with HV:
ROS formation may be derived from oxidized dietary fats. It may lead to increased protein deterioration in the liver by enhancing cell membrane LPO. Increasing ROS generation can lead to calcium homeostasis disorders, increased membrane fluidity, and cell death (Hassanen 2015). Table (5) shows no presumed divergences between R.officinalisextract remedy groups compared to standard control in CAT and MDA vigor after 30days.
Meanwhile, HV sensitivity increased by approximately 36.8 % (p > 0.05) and 42.4 % (p > 0.05) respectively, while CAT vigor increased by approximately 20.2 % and 30.4 % relative to the control group ( p > 0.05). However, the MDA scale showed a suspected diminishing (p > 0.05) in handling with R.officinalis extract simultaneously as 2 or 4 h exposure to HV. R.officinalis in rats exposed to HV for 2 or 4 h, respectively, decreasing the elevation in MDA by around 16 and 13 percent. Meanwhile, the catalase vigor scale showed a presumed increase (p>0.05) in handling with R.officinalis extract at the same time as HV exposure for 2 and 4 h R.officinalis at the same time as HV exposure for 2 and 4 h, increased CAT vigor by around 31 % and 5 % compared with HV exposure groups for 2 and 4 h, respectively. EMF extends the existence of free radicals and may function as a cancer promoter or co-promotor.
Changes in the MDA scale thus indicate increased ROS development during exposure periods, which may represent the pathological process of exposure to EMF (Ozguner et al. 2005). Plentiful studies have shown that exposure to EMF is capable of causing the body's significant oxidative degradation (Sharma et al. 2017). Other researchers reported similar observations in these parameters when rats were magnetically exposed to 50 Hz (Ibrahim et al. 2008), low-frequency electric field (Guler et al. 2008), microwave (Sokolovic et al. 2008), EMF (900 MHz) (Abd El Rahman et al. 2014), EMF (Khaki 2016). EMF led to enhanced MDA content and raised H2O2 accumulation by inducing oxidative stress and cellular deterioration (Grigor'ev et al. 2010). The electromagnetic field may be harmful by accelerating the loss of hepatocyte plasma membrane integrity (Đinđić et al. 2010). Rosemary has increased renal CAT reduction and decreased MDA scale elevation, which can be due to the antioxidant properties that inhibit LPO (Hassanen 2015). The Usage of food, which contains high antioxidants, became immensely popular since plentiful diseases have been associated with oxidative stress (Hamzaa et al. 2012). The most crucial function of antioxidants is to stimulate endogenous antioxidant defense systems or scavenge interactive species (Labban et al. 2014). The reduction in CAT's enzymatic vigor may be Because of the raised utilization of this antioxidant to counteract LPO production (Kalpana and Menon 2004). The suspected decrease in CAT vigor may be due to the excess ROS that interacts with the enzyme molecules that cause denaturation and partial inactivation (Mansour 2013).
Table 8
Effect of R.officinalis extract on liver oxidative enzymes in liver rats exposed to HV 2 and 4 h.
Groups
|
CAT (U/L)
|
MDA(nmole/g)
|
Control
|
2.63 ± 0.01
|
4.95 ± 0.04
|
R
|
2.70 ± 0.03
|
4.93 ± 0.05
|
M
|
2.71 ± 0.04
|
4.73 ± 0.04
|
MIX
|
2.80 ± 0.07
|
4.65 ± 0.02
|
HV2h
|
2.10a ± 0.06
|
6.77a ± 0.03
|
HV4h
|
1.83a ± 0.01
|
7.05a ± 0.06
|
HV2h&R
|
2.40ab ± 0.02
|
5.67ab ± 0.05
|
HV4h&R
|
2.20ab ± 0.05
|
6.17ab ± 0.02
|
Data represent the mean ± S.E. of observations from eight rats.aSignificantly different from control group at P < 0.05.bSignificantly different from HV group at P < 0.05.
|
Effect of R.officinalis extract on histological examination of rats exposed to HV for 2 and 4 h
Histological changes to help the tested biochemical markers of damage to organs were screened. It is noted that autopsy samples taken from rat liver exposed to HV for 2 or 4 h manifest several alterations such as activation of kupffer cells, central vein and sinusoid swelling, hydropic hepatocyte degeneration, and fibroplasia in a portal triad with the emergence of focal hepatic necrosis associated with inflammatory cell infiltration in a community exposed to HV for 4 h. However, handling with either R.officinalis improved hepatic histopathology, with only mild kupffer cell activation in the group exposed to HV for 2 and 4 h. concomitant with R. officinalis. (Table 6 & Fig. 1).
The ELF-EMF caused focal centrilobular necrosis of the hepatic cells surrounded by severe hydropic degeneration involving most hepatic parenchyma in liver mice (Hashem and El-Sharkawy 2009).
Table 9
Histopathological notes on liver tissue of rats exposed to HV for 2 and 4 h plus R.officinalis.
Histo-
pathological lesion
Treatment
|
Kupffer cells activation
|
Congestion of central vein and sinusoids
|
Vacuolation or hydropic degeneration of hepatocytes
|
Focal hepatic necrosis associated with inflammatory cells infiltration
|
Portal fibroplasia
|
Control
|
ـــ
|
ـــ
|
ـــ
|
ـــ
|
ـــ
|
R
|
ـــ
|
ـــ
|
ـــ
|
ـــ
|
ـــ
|
HV2h
|
+
|
++
|
++
|
ـــ
|
+
|
HV4h
|
++
|
++
|
++
|
ـــ
|
++
|
R&HV2h
|
++
|
ـــ
|
ـــ
|
ـــ
|
ـــ
|
R&HV4h
|
+
|
ـــ
|
+
|
ـــ
|
ـــ
|
(-) no histopathological change (+) mild histopathological changes
(++) moderate histopathological changes (+++) severe histopathological changes.
|