硒对氟中毒鸡淋巴细胞亚群及脾淋巴细胞凋亡的影响

摘要

目的 观察硒对慢性氟中毒鸡血液、脾脏CD4~+,CD8~+淋巴细胞亚群以及脾淋巴细胞凋亡的影响,并探讨其作用机制.方法 将8日龄海蓝褐雏鸡180只按体质量随机分为3组:对照组、氟中毒组、硒拮抗组,每组60只,分别饲以含氟195、1000、1000 ms/kg、含硒0.08、0.08、4.00 mg/kg的全价日粮.于30、60、90 d用流式细胞术检测外周血和脾脏中CD4~+、CD8~+淋巴细胞水平,TUNEL法检测脾淋巴细胞凋亡情况.结果 与对照组比较,30、60、90 d氟中毒组外周血CD4~+淋巴细胞水平降低[(35.36±4.27)%vs(24.29±2.96)%、(47.65±5.42)%vs(41.62±3.96)%、(49.58±3.98)%vs(42.35±6.03)%,P<0.05或<0.01],CD4~+/CD8~+比值也明显降低[(1.701±0.145)%vs(1.393±0.163)%、(2.712±0.345)%vs(1.781±0.201)%、(2.438±0.356)%vs(1.973 ±0.229)%,P<0.05或<0.01];与氟中毒组比较.30、60、90 d硒拮抗组外周血中CD4~+淋巴细胞水平升高[(29.40±3.38)%、(45.40 ±6.01)%、(46.85 ±5.25)%,P<0.05或<0.01],60、90 d时CD4~+/CD8~+比值明显回升[(2.004±0.314)%、(2.211±0.229)%,P均<0.01].与对照组比较,30、60、90 d氟中毒组脾脏中CD4~+淋巴细胞水平降低[(47.33±5.35)%vs(41.91±4.83)%、(49.28±5.24)%vs(41.26 ±4.56)%、(34.31±4.15)%vs(29.33 ±2.89)%,P均<0.01],CD4~+/CD8~+比值也明显降低[(1.927±0.244)%vs(1.525±0.265)%、(1.847±0.224)%vs(1.640±0.198)%、(1.265±0.174)%vs(0.878 ±0.092)%,P<0.05或<0.01];与氟中毒组比较,60、90 d硒拮抗组脾脏中CD4~+淋巴细胞水平升高[(44.87±5.43)%、(32.62 ±3.37)%,P均<0.05],而30、60、90 d时CD4~+/CD8~+比值明显回升[(1.703 ±0.201)%、(1.772±0.215)%、(0.991±0.124)%,P<0.05或<0.01].30、60、90 d时氟中毒组鸡脾淋巴细胞凋亡指数[(2.31 ±0.36)%、(2.76±0.22)%、(3.04 ±0.29)%]明显高于对照组[(1.14 ±0.21)%、(1.23±0.23)%、(1.29 ±0.20)%],而60、90 d时硒拮抗组[(2.42 ±0.32)%、(2.73±0.39)%]低于氟中毒组,差异均有统计学意义(P<0.05或<0.01).结论 一定程度的硒摄入能减少氟中毒鸡淋巴细胞的凋亡、改善失衡的淋巴细胞亚群来拮抗氟的毒性.%Objective To study the effect of selenium on peripheral and splentic T-cell subset, apoptosis of spleen cells in fluorosis chicken and its mechanism. Methods One hundred and eighty 8-day Hailanhe chicks were randomly divided into 3 groups(each 60): ①control group: 195 mg/kg fluoride and 0.08 mg/kg of selenium; ②fluorine group : 1000 mg/kg fluoride and 0.08 mg/kg of selenium ;③selenium antagonism group : 1000 mg/kg On 30~(th), 60~(th), 90~(th) day, peripheral and splentic CD4~+, CD8~+ T-cell subset analyses underwent flow cytometry and apoptosis of spleen cells were detected by TUNEL for study subjects. Results Compared with control group, the CD4~+ T-cell subset of peripheral in fluorine group was decreased obviously in 30,60,90 days[ (35.36± 4.27)% vs (24.29 ± 2.96)%, (47.65 ± 5.42)% vs (41.62 ± 3.96)%, (49.58 ± 3.98) % vs (42.35 ± 6.03 )%, P < 0.05 or < 0.01 ], CD4~+/CD8~+ ratio also was decreased obviously [ ( 1.701 ± 0.145 )% vs (1.393 ± 0.163)%,(2.712 ± 0.345)% vs (1.781 ± 0.201)%,(2.438 ± 0.356)% vs (1.973 ± 0.229)%, P< 0.05 or < 0.01]. Compared with fluorine group, the CD4~+ T-cell subset of peripheral in selenium antagonism group [ (29.40 ± 3.38)%, (45.40 ± 6.01 )%, (46.85 ± 5.25)%, P < 0.05 or < 0.01 ] was increased obviously in 30,60,90 days,CD4~+/CD8~+ ratio in 60,90 days[(2.004 ±0.314)%,(2.211±0.229)%,all P<0.01]also was increased obviously.Compared with control group,the CD4~+ T-cell subset of spleen cells in fluorine group was decreased obviously in 30,60,90 days[(47.33±5.35)% vs(41.91±4.83)%,(49.28±5.24)% vs(41.26 ±4.56)%,(34.31±4.15)% vs(29.33±2.89)%,all P<0.01],CD4~+/CD8~+ ratio also was decreased obviously[(1.927 ±0.244)% vs(1.525 ±0.265)%,(1.847±0.224)% vs(1.640±0.198)%.(1.265±0.174)% vs(0.878±0.092)%,P<0.05 or<0.01].Compared with fluorine group,the CD4~+ T-cell subset of spleen cells in selenium antagonism group in 60,90 days[(44.87±5.43)%,(32.62±3.37)%,all P<0.05]was increased obviously,CD4~+/CD8~+ ratio in 30,60, 90 days[(1.703 ±0.201)%,(1.772±0.215)%,(0.991±0.124)%,P<0.05 or<0.01]also was increased obviously. The apoptosis ratio of spleen cells in fluorine group in 30,60,90 days[(2.31±0.36)%,(2.76±0.22)%,(3.04± 0.29)%]was higher than that in control group[(1.14±0.21)%,(1.23±0.23)%,(1.29±0.20)%,P<0.01].The apoptosis ratio of spleen cells in selenium antagonism group in 60,90 days[(2.42 ±0.32)%,(2.73±0.39)%]was lower than that in fluorine group(P<0.05 or<0.01).Conclusion A certain concentration of selenium can antagonize the immunity inhibition of fluorine by decreasing apoptosis and improving the unbalance of T-cell subset.