Ann Surg Treat Res.  2018 Apr;94(4):174-182. 10.4174/astr.2018.94.4.174.

Evaluation of prophylactic and therapeutic effects of ruscogenin on acute radiation proctitis: an experimental rat model

• Yavuz E
• Karagulle
• Ercan G
• Celik A
• Yigitbas H
• Bayrak BY
• Tartar R
• Kusaslan R
• Altinel Y
• Gulcicek O

• KMID: 2407746
• DOI: http://doi.org/10.4174/astr.2018.94.4.174

Abstract

PURPOSE
Radiation proctitis (RP) is inflammation and damage to the rectum, manifested secondary to ionizing radiation utilized for treatment. In this study, we evaluated the anti-inflammatory therapeutical and protective effects of ruscogenin in a model of acute RP.
METHODS
Thirty-two Sprague-Dawley rats were divided into 4 groups (n = 8) as sham, control, treatment, and prophylaxis groups. Prophylaxis group and treatment group were dosed ruscogenin by oral gavage for 14 days pre- and postradiation. At the end of the 28th day, all subjects were sacrificed.
RESULTS
Histopathological analysis showed a significant increase in cryptitis abscess, cryptitis and reactive atypia, and depth of lymphocytic infiltration of the control group, compared to the other groups (P < 0.05), while treatment and prophylaxis groups showed significant decreases (P < 0.05). Immunohistochemical analysis indicated that immunoreactivity were significantly higher in control group (P < 0.05, P < 0.001, and P < 0.01, respectively), but vice versa for treatment and prophylaxis groups. There was not any significant difference for fibroblast growth factor 2 immunoreactivity. The epithelium of control rectums indicated an increase in TNF-α immunoreactivity while other groups had significant decrease (P < 0.01). Electron microscopical findings were parallel to light microscopy.
CONCLUSION
In this study, ruscogenin was observed to be effective on prophylaxis or treatment of acute RP. Although there are various reports on the treatment of the rectum damaged by acute RP in the literature, this could be the first study since there is no research indicating the ultrastructural effect of ruscogenin.

Keyword

Ruscogenin; Proctitis; Radiation

MeSH Terms

Abscess
Animals
Epithelium
Fibroblast Growth Factor 2
Inflammation
Microscopy
Models, Animal*
Proctitis*
Radiation, Ionizing
Rats*
Rats, Sprague-Dawley
Rectum
Therapeutic Uses*
Fibroblast Growth Factor 2
Therapeutic Uses

Figure

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  Fig. 1 Application of radiation (A, B) and dissection of rectums (C, D) after the sacrification of rats.

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  Fig. 2 Rectum samples from four experimental groups (sham, control, prophylaxis, and treatment) (H&E, ×100). (A) Sham group, (B) control (radiation) group, (C) prophylaxis group, and (D) treatment group. E, surface epithelium; C, cryptic glands; S, stroma with lamina propria; Su, submucosa; L, lumen of rectum.

• 

  Fig. 3 Immunoreactivities for FGF-2 (A–D), HIF-1α (E–H) cytokines and vascular endothelial growth factor marker (I-L) in 4 experimental groups: (A–H, ×200; I–L, ×100) (A, E, I) sham group, (B, F, J) control (radiation) group, (C, G, K) treatment group, and (D, H, L) prophylaxis group. E, surface epithelium; C, cryptic glands; S, stroma with lamina propria; Su, submucosa; L, lumen of rectum; V, vessels.

• 

  Fig. 4 Electron microscopical analysis of rectum sections from experimental rat model of radiation proctitis. (A1) On rectum surface epithelium of the sham group, microvilli (white arrow) at regular number and typical ultrastructure and tight junctions with regular ultrastructure (red arrow) were observed (×12,000). (A2) All parts of cryptic epithelium (N) of the sham group was shown to protect of their basement membrane (blue arrow) with homogenous and continuous ultrastructure, while intercellular area of the epithelium had epithelial digitations (asterisk) in accordance with normal rectum physiology (×10,000). (B1) Of the cryptic epithelial cells (E) in the control group, chromatin models of the nuclei (N) were noticed to alter, some of them showed diffuse heterochromatic appearance. Moreover, apical migration was remarkable in nuclei. Reduced number of shortened microvilli (white arrow) and intercellular digitations (asterisk) were observed both in cryptic and surface epithelia (×7,500). (B2) At basal level of surface epithelium (E) of the control group, the number of vacuoles (V) increased; number of microvilli (white arrow) decreased with highly shortened forms; the continuity and homogeneity of basement membrane (blue arrow) was lost and locally nude (green arrow). Fibrin augmentation (Fi) in the lamina propria (Lp) due to fibrosis was remarkable (×7,500). (C1) On the rectum surface epithelium of the prophylaxis group, microvilli (white arrow) projecting through lumen (L) at typical ultrastructure acquired their normal number and height. A large number of tight junctions (red arrows) also protected integrity of the epithelium (E) (×10,000). (C2) In prophylaxis group, the basement membrane (blue arrows) protected its continuity and homogeneity and the intercellular area (asterisk) had normal digitations. The Lp was remarkable to have a little increase in the extracellular matrix (×10,000). (D1) On the rectum surface epithelium of the treatment group, microvilli (white arrow) projecting through L at typical ultrastructure acquired their normal number and height. Intercellular borders (asterisk) protected their distance and a large number of tight junctions (red arrows) protected the integrity of the epithelium (E) (×10,000). (D2) The basement membrane (blue arrows) of the cryptic epithelium lost its continuity and homogeneity; abnormal digitations were observed in the intercellular spaces (red asterisk). Ultrastructure of mithochondrial (M) crista at the suprabasal locations was observed to be impaired. Normal number but irregular distribution of the fibers was remarkable at subjacent Lp (×15,000).

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