Clin Exp Otorhinolaryngol.  2017 Jun;10(2):181-187. 10.21053/ceo.2016.00262.

Effect of Hippophae rhamnoides Extract on Oxidative Oropharyngeal Mucosal Damage Induced in Rats Using Methotrexate

• Erhan E
• Terzi S
• Celiker M
• Yarali O
• Cankaya M
• Cimen FK
• Malkoc I
• Suleyman B

• KMID: 2380438
• DOI: http://doi.org/10.21053/ceo.2016.00262

Abstract

OBJECTIVES
The objective of this study is to investigate and evaluate the effect of Hippophae rhamnoides extract (HRE) on oropharyngeal mucositis induced in rats with methotrexate (MTX) through biochemical, gene expression, and histopathological examinations.
METHODS
Experimental animals were divided into a healthy group (HG), a HRE+MTX (HREM) group, HRE group (HREG), and a control group that received MTX (MTXG). The HREM and HREG groups of rats was administered 50 mg/kg HRE, while the MTXG and HG groups were given an equal volume distilled water with gavage. Then, the HREM and MTXG rat groups were given oral MTX at a dose of 5 mg/kg 1 hour after HRE and distilled water was administered. This procedure was repeated for 1 month. At the end of this period, all of the animals were sacrificed with a high dose of anesthesia. Then, the amounts of malondialdehyde (MDA) and total glutathione (tGSH) were determined in the removed oropharyngeal tissues. Interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) gene expressions were measured, and all the tissues were studied histopathologically.
RESULTS
The amount of MDA was significantly increased in the MTXG group compared to the HREM, HREG, and HG groups (P<0.001). MTX significantly decreased the amount of tGSH in the MTXG group compared to the HREM, HREG, and HG groups (P<0.001). In this study, there were no visible ulcers in the animal group in which the levels of MDA, IL-1β, and TNF-α were high and the level of tGSH was low. However, histopathologic examination revealed mucin pools in wide areas due to ruptured oropharynx glands, and proliferated, dilated, and congested blood vessels and dilated ductal structures in some areas.
CONCLUSION
HRE protected oropharyngeal oxidative damage induced by MTX. As an inexpensive and natural product, HRE has important advantages in the prevention of oropharyngeal damage induced by MTX.

Keyword

Methotrexate; Gene Expression; Hippophae; Rhamnoides; Oral Mucositis; Rat

MeSH Terms

Anesthesia
Animals
Blood Vessels
Estrogens, Conjugated (USP)
Gene Expression
Glutathione
Hippophae*
Malondialdehyde
Methotrexate*
Mucins
Mucositis
Necrosis
Oropharynx
Rats*
Stomatitis
Ulcer
Water
Estrogens, Conjugated (USP)
Glutathione
Malondialdehyde
Methotrexate
Mucins
Water

Figure

• 

  Fig. 1. The effects of Hippophae rhamnoides extract (HRE) on malondialdehyde (MDA) levels. HREM, HRE+methotrexate (MTX); MTXG, control group that received MTX; HREG, HRE group; HG, healthy group. **P<0.001.

• 

  Fig. 2. The effects of Hippophae rhamnoides extract (HRE) on total glutathione (tGSH) levels. HREM, HRE+methotrexate (MTX); MTXG, control group that received MTX; HREG, HRE group; HG, healthy group. *P<0.01. **P<0.001.

• 

  Fig. 3. The effects of Hippophae rhamnoides extract (HRE) on Interleukin-1β (IL-1β) and tumor necrosis factor (TNF-α) gene expression level. HREM, HRE+methotrexate (MTX); MTXG, control group that received MTX; HREG, HRE group; HG, healthy group. **P<0.001.

• 

  Fig. 4. Normal histopathological appearance of the healthy group oropharyngeal tissue (H&E, ×40).

• 

  Fig. 5. (A) Mucin pools in wide areas (star) due to the ruptured gland in the oropharyngeal tissue of the control group that received methotrexate (MTXG) (H&E, ×100). (B) Proliferated, dilated, and congested blood vessels (arrow) in the oropharyngeal tissue of the MTXG group (H&E, ×100). (C) Dilated ductal structures in the oropharyngeal tissue of the MTXG group (H&E, ×100). (D) Inflamed area (arrow) containing significant polymorphonuclear leukocytes in the oropharyngeal tissue of MTXG group (H&E, ×40).

• 

  Fig. 6. Appearance of Hippophae rhamnoides extract+methotrexate oropharyngeal tissue resembling healthy tissue (H&E, ×40).

• 

  Fig. 7. Normal histopathologic appearance of the oropharyngeal tissue of Hippophae rhamnoides extract group (H&E, ×40).

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