Biochemical biomarkers

Additionally, parts of the testicular tissue dried at 80ºC overnight. After recording the dry weight, samples

Significant increase of testicular MDA, TNF-α, NF-

were digested in 30 H 2 O 2 and 70 HNO 3 (1:1).

κB p65, and (iNOS), together with significant reductions

Determination of arsenic ion level at 193.69 nm took

of MDA, GSH and SOD observed in rats received

place using a spectrometry of a coupled plasma optical

NaAsO 2 in compared to the corresponding values of

emission (Optima 2100 DV, PerkinElmer, USA).

the controls (p < 0.05). Treatment with BN resulted in significant decreases of TNF-α, NF-κB p65 and (iNOS).

histopathology

(Figure1) and significant increases of GSH, MDA and

Fixing left testis took place using Bouin’s solution

Indian Journal of Forensic Medicine Toxicology, April-June 2018, Vol. 12, No. 2 258

SOD in rat testes in comparison to NaAsO 2 group that

was not treated with BN (p < 0.05) (Figure2).

histopathology and immunohistochemistry

Figure 3 demonstrates seminiferous tubular cells’ necrosis, desquamation of the epithelium and

vacuolization from NaAsO 2 administration. BN

treatment caused marked protection of the testes and

minimized the damage induced by NaAsO 2 (Figure4).

Moreover, BN significantly preserved spermatogenesis,

in comparison with NaAsO 2 group, which did not

receive BN.

The significant increase in caspase-3 expression of

Figure 3. Photomicrographs of rat testes (hE),(A, 200×)

testes for rats receiving NaAsO

2 as regard the control

from Arsenic (As) + carboxymethylcellulose (CMC) group

showing necrosis of seminiferous tubular cells, epithelial

group was demonstrated in (Figure5). Contrarily, BN

desquamation (black arrow) and vacuolization (white

administration caused a significant reduction of testicular

arrow).

tissue caspase-3 expression in comparison with NaAsO 2

group who was not treated with BN (p < 0.05).

Baicalin resulted in a significant decrease of testicular injury score (p < 0.05), and a significant increase of Johnsen score (p < 0.05) compared to the

corresponding values of NaAsO 2 non-BN treated group

(Figure 6).

Figure 4 .Photomicrographs of rat testes (hE),(A, 200×) from.Baicalin (BN) + As group showing marked improvement with minimal damage.

Figure 1. Effects of baicalin (BN) on tumor necrosis factor-α (TNF-α), nuclear factor-κB p65 (NF-κB p65), and inducible nitric oxide synthase (iNOS).

Figure 5. Immunohistochemistry (200×) of caspase-3 from Arsenic (As) + carboxymethylcellulose (CMC) group showing a significant elevation of caspase-3 immunostaining.

Figure 2. Effects of baicalin (BN) on malondialdehyde (MDA), reduced glutathione (GSh), and superoxide

259 Indian Journal of Forensic Medicine Toxicology, April-June 2018, Vol. 12, No. 2

can be attributed to the decreased nuclear translocation of NF-κB p65 observed with BN treatment.

Moreover, Caspase-3 is one of the main indicators of a cell’s entry into the apoptotic death, as it causes DNA degradation, and chromatin margination. (27-29)

Although, the metal-chelating effect of BN was detected in previous investigations, (30) the reduction in testicular tissue level of arsenic ion induced by BN in this work was statistically insignificant. This

Figure 6. Testicular injury score and spermatogenesis score .

may be explained as a higher dose of BN or a longer

DISCUSSION

experimental study may be required than that used in the present study.

The present investigation showed that BN (10 mgkgday, for 7 days) inhibited membrane lipid

Histopathology of rat testes showed major

peroxidation, as it significantly decreased MDA

seminiferous tubular cell necrosis, epithelial

production, and maintained antioxidant defenses

desquamation, loss of spermatogenesis, interstitial

(SOD activity) in rat testes exposed to arsenic toxicity.

edema, and congestion following arsenic exposure.

BN also down regulated the inflammatory responses

Similar observations were reported following arsenic

as evidenced by reduced levels of the major pro-

intoxication. (22, 25) The current study revealed BN

inflammatory mediator, TNF-α in testicular tissue.

ameliorate testicular tissue affection and significantly

Moreover, BN significantly inhibited iNOS activity in

preserved spermatogenesis in arsenic-exposed rat testes.

rat testes denting the suppression of nitrative stress. This goes in accordance with previous reports, that

In conclusion, BN may significantly have a protective

indicated the anti-oxidative, anti-nitrosative, and anti-

effect on rat testes against arsenic-induced damage. The

inflammatory properties of BN. (14-16) .Some reported the

anti-inflammatory, antioxidative, antinitrative, and anti-

Oxidativenitrative stress, increase generation of ROS

apoptotic activities of BN are the possible mechanisms.

and RNS, and inflammatory responses to be directly

Conflict of Interest: None

related to arsenic-induced damage of testes. (22, 23) Recent studies revealed that BN acts as a scavenger of ROS, and

Statement of Human and Animal Rights

RNS, prevents peroxidation of cell and mitochondrial

Institutional and national guides for the care and

membranes, and arrest the progression of inflammatory

use of laboratory animals (environment, housing and

cascades responsible for testicular tissue injury.

management) were followed.

Moreover, exposure to arsenic is known to activate

Ethical Clearance: The Research Ethics

the signaling pathway of NF-κB, resulting in transcription

Committee, King Faisal University, approved the study

of TNF-α, and iNOS genes. (24) Suppression of TNF-α

proposal (approval number: 150097)

and NF-κB significantly attenuated the arsenic-inducing tissue damage in previous investigations. (25) The NF-κB

Source of Funding: Deanship of Scientific

p65 unit is crucially sequestered in the cytoplasm, as it

Research, King Faisal University, Saudi Arabia.

binds IκB proteins to be inactivated. Increased generation of ROS and TNF-α cause rapid degradation of IκBs

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DOI Number: 10.59580973-9130.2018.00113.5

Biochemical biomarkers