The protective effects of Allium ampeloprasum Subsp Iranicum on cyclophosphamide-induced immunosuppression in NMRI Mice: A promising natural immunomodulator

Abstract

Objective: Cyclophosphamide (CTX) is an effective anticancer drug, but toxic effects against normal human tissues are its dose-limiting drawback. The aim of this research was to investigate the in vivo immunomodulatory activities of Allium ampeloprasum Subsp Iranicum against CTX-induced toxicity in mice.

Materials and methods: Extract of the whole plant of A. ampeloprasum Subsp Iranicum was obtained using the maceration technique, and its total phenolic and flavonoid contents were quantified through spectrophotometric analysis. Mice received daily oral administration (PO) of the extract (150 mg/kg) for a duration of 14 days, either as a standalone treatment or in conjunction with an intraperitoneal (IP) injection of 20 mg/kg CTX. The effects of the extract on body weight, spleen weight, white blood cell (WBC), serum antibody titer hemagglutination (HA), delayed-type hypersensitivity response (DTH), lymphocyte proliferation, cytokine production, and histopathological examinations were evaluated.

Results: A. ampeloprasum Subsp Iranicum restored several parameters including spleen weight (p<0.001), WBC (p<0.001), lymphocytes (p<0.05), and monocytes (p<0.01), HA titer (p<0.05), and DTH response (p<0.01). A. ampeloprasum Subsp Iranicum notably stimulated lymphocyte proliferation to PHA (p<0.01) and LPS (p<0.05) and recovered interferon (IFN)-γ (p<0.001) and interleukin (IL)-4 (p<0.001) levels in the immunosuppressed mice. Also, CTX-induced histopathological changes were reversed by A. ampeloprasum Subsp Iranicum .

Conclusion: Analyses revealed A. ampeloprasum Subsp Iranicum could regulate immunity and increase host immune responses. The observed strengthening effect can be attributed to the high amount of flavonoids and dipropyl trisulfide compounds present in A. ampeloprasum Subsp Iranicum.

Keywords: Allium ampeloprasum; Cyclophosphamide; Immunomodulatory; Subsp Iranicum.

Figure 1

The effects of A. ampeloprasum Subsp Iranicum extract on mice body weight before (A) and after (B) treatment. Data are expressed as the mean±SD (n=5). *p<0.05 and ***p<0.001 compared to control group (NS). Data were analyzed through one-way ANOVA coupled with Tukey-Kramer multiple comparisons test. NS (Normal Saline), CTX (Cyclophosphamide).

Figure 2

The effects of A. ampeloprasum Subsp Iranicum extract on spleen weight in CTX exposed mice. Data are expressed as the mean±SD (n=5). *p<0.05 and ***p<0.001 compared to control group (NS), +++p<0.001 compared to CTX group. Data were analyzed through one-way ANOVA coupled with Tukey-Kramer multiple comparisons test. NS (Normal Saline) and CTX (Cyclophosphamide).

Figure 3

The effects of A. ampeloprasum Subsp Iranicum extract on spleen cell number in CTX-exposed mice. Data are expressed as the mean±SD (n=5 animals). *p<0.05 compared to control group (NS), ++p<0.01 compared to CTX group. Data were analyzed through one-way ANOVA coupled with Tukey-Kramer multiple comparisons test. NS (Normal Saline) and CTX (Cyclophosphamide).

Figure 4

The effects of A. ampeloprasum Subsp Iranicum extract on hemagglutination (HA) titer in CTX-exposed mice. Data are expressed as the mean±SD (n=5). **p<0.01, ***p<0.001 compared to control group (NS), +p<0.05 compared to CTX group. Data were analyzed through one-way ANOVA coupled with Tukey-Kramer multiple comparisons test. NS (Normal Saline) and CTX (Cyclophosphamide).

Figure 5

The effects of A. ampeloprasum Subsp Iranicum extract on delayed-type hypersensitivity (DTH) response after 24 (A) and 48 hr (B) in CTX-exposed mice. Data are expressed as mean±SD (n=5). *p<0.05 compared to control group (NS), + p<0.05 and ++p<0.01 compared to CTX group. Data were analyzed through one-way ANOVA coupled with Tukey-Kramer multiple comparisons test. NS (Normal Saline) and CTX (Cyclophosphamide).

Figure 6

The effects of exposure to A. ampeloprasum Subsp Iranicum on lymphocyte proliferation response to LPS (A) and PHA (B) in CTX-exposed mice. Data are expressed as mean±SD (n=5). *p<0.05 compared to control group (NS), +p<0.05 and ++p<0.01 compared to CTX group. Data were analyzed through one-way ANOVA coupled with Tukey-Kramer multiple comparisons test. NS (Normal Saline), CTX (Cyclophosphamide).

Figure 7

The effects of A. ampeloprasum Subsp Iranicum extract on spleen histopathology after 24 hr in mice. Data are expressed as the mean±SD (n=5). NS (Normal Saline) and CTX (Cyclophosphamide).