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Modification of gamma radiation induced response of peritoneal macrophages and splenocytes by Hippophae rhamnoides (RH‐3) in mice

Alcoholic extract of Hippophae rhamnoides, RH‐3, reported to render >80% survival against lethal whole body Co‐60‐gamma irradiation (10 Gy) in mice, was investigated for its immunostimulatory effects. In comparison with un‐irradiated control, whole body irradiation did not reduce peritoneal macrophage counts at 24 h post‐irradiation. RH‐3 treatment (30 mg kg−1 body weight) alone or 30 min before whole‐body irradiation enhanced viable counts of macrophages significantly (P≤0.05) compared with both un‐irradiated control and irradiated groups. Whole‐body irradiation reduced the number of viable splenocytes significantly (P≤0.05) compared with un‐irradiated control at 24 h post‐irradiation. RH‐3 treatment alone or before whole‐body irradiation appreciably countered radiation‐induced decrease in splenocyte count. 3H‐thymidine uptake method revealed that whole‐body irradiation reduced splenocyte proliferation significantly (159 ± 45 counts min−1/106 cells; P±0.05) in comparison with control (607 ± 142 counts min−1) at 24 h after irradiation but RH‐3 treatment before irradiation reduced the steep decrease and maintained it as 444 ± 153 counts‐min−1. After whole‐body irradiation, the ratio of spleen weight/mouse weight decreased to 1.5 ± 04 compared with 2.9 ± 0.32 in un‐irradiated control at 24 h post‐irradiation. Similarly, total protein content in splenocytes also decreased to 48 ± 6 μg/106 cells in comparison with 368 ± 16 μg/106 cells of un‐irradiated control. RH‐3 treatment before irradiation countered radiation‐induced decrease in both spleen weight/mouse weight ratio (4.0 ± 0.35) and total protein content (360 ± 13 μg/106 splenocytes). In the supernatant of peritoneal macrophage cultures exposed to 2 Gy Co‐60‐gamma radiation ex‐vivo, the total nitrite content was enhanced significantly (P<0.05) to 5.72 ± 0.09 μm in comparison with un‐irradiated control (1.64 ± 0.09 μm). RH‐3 treatment (30 μg mL−1) before irradiation reduced total nitrite significantly (0.93 ± 0.3; P≤0.05) in comparison with irradiated control group. At 24 h after whole body irradiation, the CD4+/CD8+ ratio reduced to 1.5 in comparison with un‐irradiated control (1.9) but RH‐3 treatment before irradiation restored the ratio to 2.1. These findings explicitly reveal the immunostimulatory activity of RH‐3, which may play an important role in the manifestation of its radioprotective efficacy.