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Protective Effect of DPPD on Mercury Chloride-Induced Hepatorenal Toxicity in Rats.

1. Introduction

Mercury is one of the most toxic metals responsible for environmental pollution [1]. Exposure to mercury in any of its forms in different ways such as water, air, soil, and food poses serious threats to our health and the environment [2]. Following exposure, mercury ions are taken up by and accumulate in numerous organs, including the brain, intestine, kidney, liver, and placenta [3]. Based on the available experimental data, it is a reasonable hypothesis that mercury toxicity involves oxidative stress, inflammation, and apoptosis [4]. Hg[Cl.sub.2], as one of the most toxic salts of mercury, is metabolized primarily in the liver and, then, accumulated in the kidneys. Consequently, the liver and kidneys are considered the most affected organs [5]. Hg[Cl.sub.2] demolishes free radical scavenging systems such as superoxide dismutase and catalase [6], as well as increase reactive species levels that lead to disturbance of the prooxidant-antioxidant balance system causing a condition of oxidative stress [7].

N N'-diphenyl-1, 4-phenylenediamine, a grey or dark grey powder, is used as an antioxidant in rubber and oils, especially for tires in industry due to its colour and stability [8]. DPPD is one of the most frequently used and potent antioxidants. It is effective at very low concentrations and believed to be rather selective [9]. DPPD, acting as an intracellular antioxidant, enlarges the pool size of lipid-soluble antioxidants, especially in the cytoplasmic membranes, and prevents the formation of lipid peroxides resulting in maintenance of the normal mitochondrial structure and enzyme activity [10]. The antioxidant activity of DPPD implemented by the donation of hydrogen to radical derivatives breaking the autocatalytic cycle protecting cells from oxidative stress [11] suppresses necrosis and decreases reactive oxygen species (ROS) formation [12] Also, DPPD inhibits lipid peroxidation and nephrotoxicity [13]. Thus, DPPD inhibits collagen deposition, dampens apoptosis, and prevents histopathological damages [14]. The present study reports the antifibrotic effect of DPPD against hepatorenal fibrosis induced by Hg[Cl.sub.2] in rats.

2. Materials and Methods

2.1. Chemicals and Reagents. All chemicals and reagents were of the highest purity grade. DPPD (>99.8%) and Hg[Cl.sub.2] (>99.5%) were obtained from Sigma-Aldrich Chemical Company (St. Louis, MO, USA). In addition to serum ALT, AST, and ALP activities, urea, uric acid, and creatinine levels were determined using colourimetric diagnostic kits (Bio-diagnostic, Cairo, Egypt) according to the manufacturer's instructions. TGF-[beta]%, CD4%, and CD8% were analyzed by using an Accuri C6 flow cytometer (BD Biosciences, San Jose, CA). Data were quantified with C Flow software (BD Accuri, San Jose, CA). The hydroxyproline content is measured by ELISA as an important index reflecting the degree of kidney and liver fibrosis.

2.2. Animals and Experimental Design. Rats were assigned to groups by using the Statistical Package of Social Science (SPSS) program for Windows (Standard version 21). Thirty female Sprague Dawley rats, weighing approximately 170-220 gm, were purchased from the Medical Experimental Research Center (MERC), Faculty of Medicine, Mansoura University, Mansoura, Egypt. The animals were kept in polypropylene cages under standard laboratory conditions of relative humidity (45 [+ or -] 5%) and temperature (25 [+ or -] 2[degrees]C) with 12 h light/dark cycle and provided with food pellets and tap water ad libitum. Principles of laboratory animals caring (NIH publication no. 85-23, that revised 1985) were followed. Ethical protocols for laboratory animal care and use were approved and followed under the supervision of Faculty of Postgraduate Studies for Advanced Sciences (PSAS), Beni-Suef University, Experimental Animals Ethical Committee (No. BSU/EAEC/PSAS/16/112018).

Rats were randomly divided into 3 groups (10 rats/ group). The Hg[Cl.sub.2] dose was 4mg/kg, i.p.:

Group I (control): rats received saline i.p. for 14 days and served as the control

Group II (Hg[Cl.sub.2]): rats were injected with a single dose of Hg[Cl.sub.2] (4 mg/kg, i.p.) at day one of the experiment Group III (Hg[Cl.sub.2] + DPPD): rats were injected with a single dose of Hg[Cl.sub.2] (4mg/kg, i.p.) at day one of the experiments and, then, treated with DPPD (0.5 g/kg, i.p.) according to [15] once every two days starting from day 3 of the experiment

2.3. Collection and Preparation of Samples. All rats were exposed to sevoflurane anesthesia and killed by decapitation (24 h after the last injection), and urine and blood samples were collected from each rat after 14 days of Hg[Cl.sub.2] (or saline) injection. The liver and kidneys tissues were dissected and used for biochemical, flow cytometry, and histopathological examinations.

2.4. Liver and Renal Markers. To assess liver functions, serum ALT, AST, and ALP enzyme activities were detected. Also, serum urea, uric acid, and creatinine levels were determined to assess kidney functions using colourimetric diagnostic kits (Biodiagnostic, Cairo, Egypt) according to the manufacturer's instructions.

2.5. Oxidative Stress Evaluation. The activities of the antioxidant enzymes SOD, CAT, and GSH in addition to the MDA content in liver and kidney tissues were all measured using commercial laboratory diagnostic kits (Biodiagnostic Co., Cairo, Egypt).

2.6. CD8%, CD4%, and TGF-j3% Measurements. Flow cytometry detection of CD8%, CD4%, and TGF-[beta]% depends on the specific binding of monoclonal antibodies to the antigenic determinants. The monoclonal antibodies labelled with different fluorochromes which are excited via a laser beam from a flow cytometer during analysis. The fluorescence intensity differences were proportional to the expression of the analyzed antigens. Assays were analyzed by using an Accuri C6 flow cytometer (BD Biosciences, San Jose, CA). Data were quantified with C Flow software (BD Accuri, San Jose, CA).

2.7. Measurement of the Hydroxyproline Content. About 50 mg of kidney or liver tissue specimens were hydrolyzed, and then, chloramine Tsolution was added to the specimen's supernatant and, then, incubated, followed by Ehrlich's solution addition. The final mixture was incubated, and the optical density was estimated at 560 nm [16]. Hydroxyproline values were expressed as ug/mg tissue.

2.8. Histopathological Examination. The formalin-embedded liver and kidney tissues were cut into 4 [micro]m thick sections, and then, the slides were stained with hematoxylin and eosin (H&E) for histological evaluation and Masson trichrome to assess collagen deposition. We routinely conduct H&E staining to grade tubular damage (0, no damage; 1, 0-25% damaged tubules; 2, 25-50% damaged tubules; 3, 50-75% damaged tubules; and 4, >75% damaged tubules) [17] and the liver injury score of fibrosis as described by [18]. The sections were examined and photographed using an Olympus light microscope (Olympus BX51, Tokyo, Japan) with an attached digital photograph machine (Olympus E-330). Images were captured from each section randomly, and semiquantitative analysis of the fibrotic area was performed on an Intel[R] Core I5[R]-based computer using Image J software with a specific built-in routine for stain quantification and automated area measurement. Five slides were prepared from each group, 5 random fields from each slide analyzed as previously reported [19, 20].

2.9. Statistical Analyses. Data were analyzed using SPSS software version 22 for Windows (IBM, Armonk, NY, USA). Descriptive statistics were calculated in the form of Mean [+ or -] Standard deviation (SD). ANOVA and Tukey's post hoc tests were used for comparison between groups. A level of p < 0.05 was defined as statistically significant.

3. Results

3.1. Liver and Kidney Functions. The potential effects of Hg[Cl.sub.2] and DPPD treatment on renal and liver function parameters are summarized in Tables 1 and 2. The presented data showed that serum ALT, AST, ALP, creatinine, urea, and uric acid levels were significantly (p < 0.001) increased in Hg[Cl.sub.2]-injected rats compared to control and Hg[Cl.sub.2] + DPPD-treated rats. Conversely, animals treated by Hg[Cl.sub.2] + DPPD reversed all parameters' alterations towards the normal ranges.

3.2. Lipid Peroxidation and Antioxidant Enzyme Activities. The data of lipid peroxidation, CAT, GSH, and SOD activities in the renal and hepatic tissues are shown in Tables 1 and 2. Compared with control and Hg[Cl.sub.2] + DPPD group values, the Hg[Cl.sub.2] group showed a significantly (p < 0.001) increased MDA level and significantly decreased antioxidant enzymes (CAT, GSH, and SOD) activities. These results indicate that DPPD ameliorates the Hg[Cl.sub.2]-induced oxidative stress in the liver and kidney.

3.3. Hepatorenal Fibrosis Induced by Hg[Cl.sub.2] in Rats. The hydroxyproline content is a specific marker for collagen deposition. The Hg[Cl.sub.2] group showed a significantly (p < 0.001) increased hydroxyproline content in renal and liver tissues compared with the control group. In contrast, DPPD treatment significantly decreased the renal and liver hydroxyproline content (Tables 1 and 2).

3.4. Flow Cytometry. As shown in Tables 1 and 2, the TGF-[BETA], CD4, and CD8 percent showed a significant (p < 0.001) increase in Hg[Cl.sub.2]-treated rats compared to control and Hg[Cl.sub.2] + DPPD-treated rats. Conversely, animals treated by DPPD reversed TGF-[BETA], CD4, and CD8 percent alterations towards the normal ranges.

3.5. Histopathological Analysis. H&E-stained and Masson trichrome-stained kidney sections are shown in Figure 1 and liver sections are shown in Figure 2. H&E histopathological stain in the control group showed normal kidney morphology (Figure 1(a)) and normal hepatic lobular architecture with distinct hepatocytes (Figure 2(a). Hg[Cl.sub.2]-treated animals showed tubular dilatation with many degenerated signs (Figure 1(b)) and hepatic degeneration with large areas of extensive cell necrosis (Figure 2(b)). Treatment with DPPD significantly attenuated the pathological changes in both kidney (Figure 1(c)) and liver (Figure 2(c)) tissues compared to the Hg[Cl.sub.2]-treated group. Assessment of kidney (Figure 1(g)) and liver (Figure 2(g)) injury by a semiquantitative scoring system from 0 to 5. Data were mean [+ or -] SD. * p < 0.01 vs. control, # p < 0.01 vs. Hg[Cl.sub.2].

The collagen content was assessed by Masson's trichrome stain, and the control group showed a normal collagen content in the kidney (Figure 1(d)) and liver (Figure 2(d)) tissues. Significant amounts of collagen deposition were observed in the kidney (Figure 1(e)) and liver (Figure 2(e)) tissues of Hg[Cl.sub.2]-treated animals. Conversely, oadministration of DPPD + Hg[Cl.sub.2] showed a significant (p < 0.001) modulation in the collagen content level towards normal in both renal (Figure 1(f)) and hepatic (Figure 2(f)) tissues. Treatment with DPPD significantly (p < 0.001) decreased both the renal (Figure 1(h)) and hepatic (Figure 2(h)) Masson% area.

4. Discussion

Hg[Cl.sub.2] generates free radicals and subsequently increases oxidative stress, which leads to nephrotoxicity and accelerates hepatotoxicity [21]. This adverse effect of Hg[Cl.sub.2] could be eliminated by DPPD treatment probably because of its strong free radical scavenging activity through the electron(s) donation pathway, protecting cells from oxidation and necrosis [11].

In the present study, liver and renal functions were detrimentally altered after Hg[Cl.sub.2] administration causing hepatorenal dysfunction evidenced by a significant elevation in AST, ALT, and ALP enzyme activities and urea, uric acid, and creatinine levels. Similar results were reported by [22, 23]. Treatment with DPPD showed a marked improvement that was clear in the previously listed parameters, and these findings agreed with those of Kawai et al. [13] who reported that DPPD may possess an antioxidative behavior.

Hg[Cl.sub.2] administration initiates the formation of highly reactive substances such as reactive oxygen species in addition to the stimulation of oxidative stress [24]. Consequently, the lipid peroxidation level increased and the antioxidant enzymes activities decreased.

In the present study, we found that Hg[Cl.sub.2] significantly diminishes the activities of the antioxidant enzymes SOD and CAT in addition to GSH in kidney and liver tissues, whereas the end product of lipid peroxidation (MDA contents) was significantly increased compared with the control group. A variety of experiments have demonstrated parallel results [23, 25]. Conversely, coadministration of DPPD + Hg[Cl.sub.2] showed a significant modulation in the activities of SOD and CAT in addition to the level of GSH and MDA towards normal. The hepatorenal protective activity of DPPD was observed in the previous studies [12, 26].

Hydroxyproline is used for the estimation of the collagen content, considering that collagen contained 12.7% hydroxyproline by weight [27]. Our results were parallel to those of Yuan et al. [28] who concluded that renal and liver fibrosis are induced by Hg[Cl.sub.2], demonstrated by a significant elevation (p [less than or equal to] 0.001) of the hydroxyproline content in liver and kidney tissues compared to control. Consequently, treatment with DPPD significantly attenuated hydroxyproline and collagen deposition (p < 0.001) in animals [19].

Our results reported a significant increase in both renal and hepatic TGF-[BETA]%, CD4%, and CD8% in Hg[Cl.sub.2]-treated rats compared to (control and Hg[Cl.sub.2] + DPPD groups). The elevated CD4+ and CD8+ percentage may be related to the abnormal immune status due to Hg[Cl.sub.2] toxicity. These results are in agreement with the previous findings by Liu et al. [29], who indicate significant increases in CD4% and CD8% as a result of mercury induction of T-cell autoimmune syndrome, including autoantibodies and increases in TGF-[BETA] production and various other cytokines cause collagen deposition. Moreover, previous studies [30, 31] reported a significant increase in renal and hepatic TGF-[BETA]% in Hg[Cl.sub.2]-treated rats compared to control, but rats treated with Hg[Cl.sub.2] + DPPD showed a significant decrease in TGF-[beta]%, CD4%, and CD8% towards normal.

In the present study, we performed a histopathological examination to further support the biochemical and immunological evidence. We compared the morphological structure among each group using H&E stain. In the control group, there were no injuries or histological changes detected in the kidneys (Figure 1(a)) or liver (Figure 2(a)). The Hg[Cl.sub.2] group showed liver necrosis, swelling, and structure changes (Figure 2(b)) in addition to renal tissue damage, collagen formation, and atrophy in the normal tubular architecture (Figure 1(b)) compared with (control and Hg[Cl.sub.2] + DPPD groups). Treatment with DPPD significantly attenuated the pathological changes in both kidney (Figure 1(c)) and liver (Figure 2(c)) tissues. Earlier studies [4, 32] demonstrated parallel results.

Masson's trichrome stain was used for distinguishing collagen deposition. The control group showed a normal collagen content in the kidney (Figure 1(d)) and liver (Figure 2(d)) tissues, while in the Hg[Cl.sub.2] group, the collagen content was increased significantly in the kidney (Figure 1(e)) and liver (Figure 2(e)). Our results agreed with those in [33, 34]. The recorded hepatorenal lesions and collagen content in Hg[Cl.sub.2] + DPPD cotreated rats decreased significantly (p < 0.001) compared with Hg[Cl.sub.2]-treated rats (Figure 1(f)) and (Figure 2(f)). Similar protective effects for DPPD were previously reported in [13].

5. Conclusions

Finally, we conclude that the antioxidant DPPD can retard the progression of hepatorenal fibrosis and collagen deposition induced by Hg[Cl.sub.2]. Further studies are needed to explain the intrinsic and extrinsic pathways of DPPD antifibrotic efficacy.

https://doi.org/10.1155/2020/4127284

Data Availability

The data used to support the findings of this study are available from the corresponding author upon request.

Conflicts of Interest

The authors declare that they have no conflicts of interest.

References

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Ahmed Nabil [(ID)], (1,2) Mohamed M. Elshemy, (3) Medhat Asem, (3) and Heba F. Gomaa (4,5)

(1) Research Center for Functional Materials, National Institute for Materials Science (NIMS), 1-1 Namiki, Tsukuba, Ibaraki 305-0044, Japan

(2) Biotechnology and Life Sciences Department, Faculty of Postgraduate Studies for Advanced Sciences (PSAS), Beni-Suef University, Beni-Suef, Egypt

(3) Faculty of Science, Menoufia University, Menoufia, Egypt

(4) Zoology Department, Faculty of Science, Ain-Shams University, Cairo, Egypt

(5) Biology Department, Faculty of Sciences and Arts-Scientific Departments, Qassim University, Buraydah, Saudi Arabia

Correspondence should be addressed to Ahmed Nabil; drnabil_100@psas.bsu.edu.eg

Received 13 January 2020; Revised 16 April 2020; Accepted 18 June 2020; Published 15 July 2020

Academic Editor: Orish Ebere Orisakwe
Table 1: Kidney injury parameters. Values are
expressed as M [+ or -] SD of 10 animals in each group.

Variables                                   Control group

Serum:
  Creatinine (mg/dl)                     0.46 [+ or -] 0.03
  Urea (mg/dl)                          27.83 [+ or -] 3.60
  Uric acid (mg/dI)                      2.57 [+ or -] 0.25
Homogenate:
  Hydroxyproline (ug/mg tissue)         22.58 [+ or -] 0.63
  MDA (mmol/g tissue)                   59.79 [+ or -] 4.99
  SOD (U/mg protein)                    11.66 [+ or -] 0.64
  CAT (mol/min/gm)                       0.66 [+ or -] 0.09
  Glutathione ([micro]mol/g protein)    32.13 [+ or -] 2.07
  TGF-[beta] (ng/ml)                    33.27 [+ or -] 3.43
  CD4 (ng/ml)                           21.42 [+ or -] 1.04
  CD8 (ng/ml)                           23.62 [+ or -] 3.88

Variables                              Hg[Cl.sub.2] group

Serum:
  Creatinine (mg/dl)                   1.53 [+ or -] 0.21 (a)
  Urea (mg/dl)                        73.43 [+ or -] 5.87 (a)
  Uric acid (mg/dI)                    4.53 [+ or -] 0.76a
Homogenate:
  Hydroxyproline (ug/mg tissue)       41.23 [+ or -] 9.25 (a)
  MDA (mmol/g tissue)                 92.25 [+ or -] 13.05 (a)
  SOD (U/mg protein)                   6.71 [+ or -] 1.51 (a)
  CAT (mol/min/gm)                     0.35 [+ or -] 0.05 (a)
  Glutathione ([micro]mol/g protein)  21.71 [+ or -] 6.22 (a)
  TGF-[beta] (ng/ml)                  49.20 [+ or -] 9.10 (a)
  CD4 (ng/ml)                         43.70 [+ or -] 5.31 (a)
  CD8 (ng/ml)                         44.06 [+ or -] 6.77 (a)

Variables                             Hg[Cl.sub.2] + DPPD group

Serum:
  Creatinine (mg/dl)                   0.57 [+ or -] 0.10 (b)
  Urea (mg/dl)                        32.23 [+ or -] 2.30 (b)
  Uric acid (mg/dI)                    3.16 [+ or -] 0.98 (a,b)
Homogenate:                           30.24 [+ or -] 5.23 (a,b)
  Hydroxyproline (ug/mg tissue)
  MDA (mmol/g tissue)                 70.62 [+ or -] 9.69 (b)
  SOD (U/mg protein)                   9.63 [+ or -] 1.48 (a,b)
  CAT (mol/min/gm)                     0.58 [+ or -] 0.09 (b)
  Glutathione ([micro]mol/g protein)  28.21 [+ or -] 3.51 (b)
  TGF-[beta] (ng/ml)                  38.60 [+ or -] 9.19 (b)
  CD4 (ng/ml)                         34.30 [+ or -] 5.01 (a,b)
  CD8 (ng/ml)                         25.66 [+ or -] 3.87 (b)

SD: standard deviation; P: probability; * significance <0.05;
** high significance. The test used is one-way ANOVA
followed by post hoc Tukey. ** Significance relative to
the control group compared with Hg[Cl.sub.2] and
Hg[Cl.sub.2] + DPPD groups. Significance between the
Hg[Cl.sub.2] group and Hg[Cl.sub.2] + DPPD group.

Table 2: Liver injury parameters. Values
are expressed as M [+ or -] SD of 10 animals in each group.

Variables                                    Control group

Serum:
  ALT (U/L)                               33.30 [+ or -] 3.30
  AST (U/L)                               59.40 [+ or -] 5.12
  ALP (U/L)                              215.45 [+ or -] 5.42
Homogenate:
  Hydroxyproline (ug/mg tissue)           19.80 [+ or -] 1.84
  MDA (mmol/g tissue)                     81.83 [+ or -] 4.58
  SOD (U/mg protein)                      20.99 [+ or -] 1.75
  CAT (mol/min/gm)                         1.05 [+ or -] 0.10
  Glutathione ([micro]mol/g protein)      25.35 [+ or -] 1.50
  TGF-[beta] (ng/ml)                      32.31 [+ or -] 2.45
  CD4 (ng/ml)                             24.44 [+ or -] 2.64
  CD8 (ng/ml)                             19.73 [+ or -] 1.13

Variables                                 Hg[Cl.sub.2] group

Serum:
  ALT (U/L)                             64.70 [+ or -] 14.16 (a)
  AST (U/L)                            114.70 [+ or -] 15.96 (a)
  ALP (U/L)                            333.97 [+ or -] 32.48 (a)
Homogenate:
  Hydroxyproline (ug/mg tissue)         39.20 [+ or -] 1.77 (a)
  MDA (mmol/g tissue)                  112.72 [+ or -] 10.48 (a)
  SOD (U/mg protein)                    14.87 [+ or -] 2.65 (a)
  CAT (mol/min/gm)                       0.60 [+ or -] 0.28 (a)
  Glutathione ([micro]mol/g protein)    18.97 [+ or -] 3.10 (a)
  TGF-[beta] (ng/ml)                    50.60 [+ or -] 8.83 (a)
  CD4 (ng/ml)                           39.66 [+ or -] 1.60 (a)
  CD8 (ng/ml)                            38.5 [+ or -] 2.88 (a)

Variables                              Hg[Cl.sub.2] + DPPD group

Serum:
  ALT (U/L)                            41.80 [+ or -] 11.14 (b)
  AST (U/L)                            74.00 [+ or -] 5.43 (a,b)
  ALP (U/L)                           247.10 [+ or -] 25.91 (a,b)
Homogenate:                            24.36 [+ or -] 4.08 (a,b)
  Hydroxyproline (ug/mg tissue)
  MDA (mmol/g tissue)                  81.93 [+ or -] 4.02 (b)
  SOD (U/mg protein)                   19.94 [+ or -] 3.71 (b)
  CAT (mol/min/gm)                      0.93 [+ or -] 0.13 (b)
  Glutathione ([micro]mol/g protein)   22.24 [+ or -] 2.01 (a,b)
  TGF-[beta] (ng/ml)                   34.60 [+ or -] 4.92 (b)
  CD4 (ng/ml)                          25.30 [+ or -] 3.31 (b)
  CD8 (ng/ml)                          21.07 [+ or -] 3.84 (b)

SD: standard deviation; P : probability; * significance
<0.05; ** high significance. The test used was one-way
ANOVA followed by post hoc Tukey. (a) Significance relative
to the control group compared with Hg[Cl.sub.2] and
Hg[Cl.sub.2] + DPPD groups. (b) Significance between
Hg[Cl.sub.2] group and Hg[Cl.sub.2] + DPPD group.

Figure 1: H&E-stained and Masson-stained kidney tissues of rats
from different groups (magnification x200). (a) Section of the
control group showing the normal architecture of the kidney. (b)
Significant increase in tubular dilatation and degenerative changes
observed in Hg[Cl.sub.2]-injured rats. (c) Treatment with DPPD
significantly attenuated the renal histopathological changes. (d)
Masson's trichrome staining indicated no abnormal collagen in the
control group. (e) Sections of Hg[Cl.sub.2]-treated group indicated
an increase in fibrosis stained in blue. (f) Kidney section of rats
cotreated with Hg[Cl.sub.2] + DPPD showed a significant decrease in
collagen deposits. (g) Pathological scoring showed a significant
increase in the tubular injury score in the Hg[Cl.sub.2]-treated
group when compared with other groups. (h) Comparison between
different groups in the Masson% area. Data were mean [+ or -] SD.
* p < 0.01 vs. control; # p < 0.01 vs. Hg[Cl.sub.2].

Control               0.5
Hg[Cl.sub.2]          3.8
Hg[Cl.sub.2]+DPPD     1.9

Note: Table made from bar graph.

Figure 2: H&E-stained and Masson-stained liver tissues of rats from
different groups (magnification x200). (a) Hepatic histology of the
control group, showing normal hepatic lobular architecture. (b)
Hepatic degenerative changes with extensive cell necrosis observed
in Hg[Cl.sub.2]-injured rats. (c) Rats treated with DPPD showed a
significant modulation in the hepatic histology towards normal. (d)
Control group stained with Masson's trichrome showed that the
natural structure and collagen fibers cannot be seen. (e)
Accumulation and progression of collagen fibers in the liver of the
Hg[Cl.sub.2] group. (f) Significant decrease in collagen fibers
observed in rats cotreated with Hg[Cl.sub.2] + DPPD. (g)
Pathological scoring showed a significant increase in the hepatic
injury score in the Hg[Cl.sub.2]-treated group compared with other
groups. (h) Comparison between different groups in the Masson%
area. Data were mean [+ or -] SD. * p < 0.01 vs. control; # p < 0.01
vs. Hg[Cl.sub.2].

Control              0.3
Hg[Cl.sub.2]         3.6
Hg[Cl.sub.2]+DPPD    1.2

Note: Table made from bar graph.
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Article Details
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Title Annotation:Research Article
Author:Nabil, Ahmed; Elshemy, Mohamed M.; Asem, Medhat; Gomaa, Heba F.
Publication:Journal of Toxicology
Geographic Code:7EGYP
Date:Jul 31, 2020
Words:5092
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