【摘要】 目的:探讨家兔膀胱黏膜下注射卡介菌多糖核酸(polysaccharide nucleic acid fraction of Bacillus Calmette-Guerin,BCG-PSN)所致的免疫效应变化,并寻求最合适注射的药物剂量。方法:将实验家兔随机分成5组,分别为BCG-PSN大、中、小(0.35、0.175、0.0875 mg/ml)剂量膀胱黏膜下注射组,BCG(0.35 mg/ml)膀胱黏膜下注射对照组和BCG-PSN(0.35 mg/ml)膀胱灌注对照组。分别在用药前和用药后1周、2周、1和3个月应用流式细胞术和酶联免疫吸附测定法(ELISA)对家兔外周血中T淋巴细胞亚群(CD4~+、CD8~+)和细胞因子(IL-2、TNF-α、IFN-γ)进行测定;通过H-E、Masson染色对家兔膀胱壁组织进行病理检查。结果:(1)注射BCG-PSN各组CD4~+、CD8~+ T细胞的数量随用药剂量增加而增加,同时随时间延长逐步增加,用药后2周达到高峰,用药后3个月基本恢复至原水平;BCG注射对照组变化趋势与之相同,BCG-PSN灌注对照组的CD4~+ T细胞数量明显低于大剂量BCG-PSN注射组(P<0.05),各组的CD4~+/CD8~+变化趋势相似。(2)注射BCG-PSN各组的细胞因子(IL-2、TNF-α、IFN-γ)变化与CD4~+、CD8~+ T细胞的变化趋势相同,BCG-PSN灌注对照组的细胞因子数最明显低于大、中剂量BCG-PSN注射组(P<0.05)。(3)各用药组膀胱黏膜下均见大量淋巴细胞浸润,BCG注射对照组家兔膀胱壁组织出现纤维化改变。结论:膀胱黏膜下注射BCG-PSN所致的免疫效应与注射BCG相似,优于BCG-PSN膀胱灌注,且不引起膀胱壁纤维化;以0.35 mg/ml注射效果最明显。更多还原

【Abstract】 Objective:To investigate the immune response after injecting polysaccharide nucleic acid of Bacillus Calmette-Guerin(BCG-PSN)under the mucous membrane of bladder in rabbits and to search for the most suitable dose of BCG-PSN.Methods:The rabbits were randomly divided into 5 groups:high dose(0.35 mg/ml)BCG-PSN group,mid- dle dose(0.175 mg/ml)BCG-PSN group,low dose(0.0875 mg/ml)BCG-PSN group,BCG(0.35 mg/ml)control group and BCG-PSN(0.35 mg/ml)intra-bladder perfusion control group.T lymphocyte subsets(CD4~+,CD8~+)in the peripheral blood were determined by flow cytometry before and 1 week,2 weeks,1 month and 3 months after the treatment; the levels of IL-2,TNF-α,and IFN-γof peripheral blood were determined by enzyme-linked immunosorbent assay;H-E and Masson staining was used for the pathological examination of the bladder.Results:(1)BCG-PSN injection increased the number of CD4~+ and CD8~+ T lymphocytes in a time- and dose-dependent manner,with the peak numbers appeared 2 weeks after BCG-PSN injection;the numbers restored to the normal levels 3 months after BCG-PSN injection.BCG control group had a similar changing pattern to the BCG-PSN injection group.The number of CD4~+ T cells BCG-PSN perfusion group was significantly lower than that in the high-dose BCG-PSN injection group(P<0.05).The changing tendencies of CD4~+/CD8~+ were similar in both groups.(2)The changes of IL-2,TNF-α,and IFN-γlevels in BCG-PSN injection groups were similar to those of CD4~+ and CD8~+ T cells;and the levels in BCG-PSN perfusion group were significantly lower than those of the high-and median-dose of BCG-PSN injection groups(P<0.05).(3)T lymphocytes infihration of mucous memberance uncer bladder was observed in all groups after the treatment.The BCG injection control group had fibrosis of the bladder wall.Conclusions:The immune response of BCG-PSN injection under the mucous membrane of bladder can cause similar immune response to that of BCG;the response is superior to that of BCG-PSN perfusion;and BCG-PSN causes no bladder wall fibrosis.The optimal dose of BCG-PSN is O.35 mg/ml.更多还原