Therapeutic effect of intensity modulated radiation therapy on secondary femur infection with Echinococcus granulosus in rats

doi:10.12140/j.issn.1000-7423.2021.04.004

Abstract

Abstract:

Objective To observe the therapeutic effect of intensity modulated radiation therapy (IMRT) on secondary femur infection with Echinococcus granulosus in rats. Methods Seventy-five male Wistar rats with secondary femur infection of Echinococcus metacestode were used in this study. They were divided into the IMRT group (n = 50) and the control group (n = 25). In the IMRT group, the left thighbone of the rat was irradiated at irradiation dose of 40 Gy. The rats in the control group received no treatment. E. granulosus cysts were obtained from the left and right thighbones respectively, from which the germinal layer cells were extracted. Hoechst 33258 staining was performed to observe morphological changes of nuclei, and immunohistochemistry was conducted to detect the expression of apoptosis-related Caspase-3 and p53. ELISA was used to detect the difference of interferon γ (IFN-γ), tumor necrosis factor-α (TNF-α), interleukin-4 (IL-4) and IL-10 cytokine levels in the mice serum of IMRT group and control group. Flow cytometry was used to detect the CD4⁺/CD8⁺ ratio in splenic lymphocytes. Results One week after IMRT treatment, the absorbance (A₄₉₀ value) of Th1 cytokines IFN-γ and TNF-α in the rat serum of the IMRT group were 322.45 ± 94.42 and 28.76 ± 6.23, respectively, which were higher than those of the control group (277.82 ± 65.48 and 19.63 ± 4.76) (P < 0.05). The A₄₉₀ value of Th2 cytokine IL-4 was 11.02 ± 2.55, which was lower than that of the control group (13.22 ± 3.13) (P < 0.05). The IL-10 A₄₉₀ value was 16.57 ± 3.02, which showed no statistical difference from the control group (20.51 ± 3.17) (P > 0.05). Hoechst 33258 fluorescent staining showed apoptosis of the germinal layer cells of the left femoral hydatid cysts of the IMRT group, while the right germinal layer cells were normal. The results of immunohistochemistry showed abundant expression of Caspase-3 and p53 in the germinal layer cells of the left femoral hydatid cyst of rats in the IMRT group, but not in the right germinal cells. The results of flow cytometry showed that the CD4⁺/CD8⁺ ratio in the IMRT group (2.96 ± 1.90) was much higher than that in the control group (1.65±1.31)(P < 0.05). Conclusion IMRT can induce apoptosis of the germinal layer cells of the femoral hydatid cyst in rats. The mechanisms may involve the regulation of Caspase-3 and p53, and the radiation triggering host immune responses to the parasites.

Key words: Echinococcus granulosus, Infection, Intensity modulated radiation therapy, Femur, Apoptosis, T lymphocyte subsets

CLC Number:

R532.32

ZHOU Wen-zheng, SUN Jun-gang, ZHAO Xi-bin, CAO Li. Therapeutic effect of intensity modulated radiation therapy on secondary femur infection with Echinococcus granulosus in rats[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2021, 39(4): 443-448.

Figures/Tables 2

References 21

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