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目的探讨中药复方提取物湘A-1(Hu Nan A-1,HNA-1)对猴免疫缺陷病毒(Simian immunodeficiency virus,SIV)慢性感染中国恒河猴胸腺输出功能的影响。方法采用SIVmac239病毒液对8只中国恒河猴进行感染16~21个月,随机分为对照组和治疗组,每组4只。治疗组给予HNA-1灌胃20 m L治疗,每日1次;对照组给予等体积生理盐水灌胃。两组给药2个月。观察两组一般情况、体重,采用实时荧光定量PCR方法检测两组血浆病毒载量,采用流式细胞法检测CD4比例及数量、CD4的初始亚群,实时荧光定量PCR法检测T细胞受体DNA重排删除环(TREC),常规HE染色后观察胸腺组织的病理情况。分析胸腺组织病变情况与CD4细胞数、初始型CD4细胞数及TREC的关联性。结果治疗后两组体重、病毒载量、CD4比例绝对数比较,差异无统计学意义(P>0.05)。对照组TREC改变倍数明显呈下降趋势,治疗组总体呈明显的上升趋势,两组间差异有统计学意义(P<0.05)。两组动物的胸腺组织均可见结构破坏、粉红色无结构样物质填充、结缔组织增多、细胞密度降低、排列紊乱等病理改变,未见明显区别。TREC含量与初始型CD4去除极值后,呈正相关(r=0.926,P=0.001);而初始型CD4数量与CD4数量呈正相关(r=0.961,P=0.005)。结论检测TREC含量来测定新胸腺迁出细胞数量可作为评价胸腺输出功能的检测方法,且HNA-1可增加胸腺的输出功能,而TREC、初始型CD4数量及胸腺组织病理情况之间存在一定相关关系,在感染晚期尤为明显。
Objective To investigate the effect of Hu Nan A-1 (HNA-1), a traditional Chinese medicine extract, on thymic output of Chinese rhesus monkeys with Chronic Infection of Simian Immunodeficiency Virus (SIV). Methods 8 Chinese Rhesus monkeys were infected with SIVmac239 virus solution for 16-21 months and randomly divided into control group and treatment group with 4 in each group. The treatment group was given HNA-1 intragastric administration of 20 m L once daily; the control group was given equal volume of normal saline. Two groups were given for 2 months. The general condition and body weight of the two groups were observed. The plasma viral load of the two groups was detected by real-time fluorescence quantitative PCR. The proportion of CD4 and the number of CD4 were detected by flow cytometry. The initial subsets of CD4 were detected by real-time fluorescence quantitative PCR. Rearrangement and deletion of ring (TREC), routine HE staining after thymus tissue pathology. Analysis of thymus lesions and CD4 cell count, initial CD4 cell count and TREC association. Results After treatment, there was no significant difference in the absolute numbers of body weight, viral load and CD4 ratio between the two groups (P> 0.05). The change of TREC in the control group decreased obviously, the treatment group showed an obvious upward trend, the difference between the two groups was statistically significant (P <0.05). There was no obvious difference between the two groups of animals in their thymus structure destruction, pink non-structure-like material filling, connective tissue increase, cell density reduction and disordered arrangement. There was a positive correlation between TREC level and CD4 count (r = 0.926, P = 0.001). However, there was a positive correlation between CD4 (superscript +) and CD4 (r = 0.961, P = 0.005). Conclusion Detection of TREC content to determine the number of newly-released thymus cells can be used as a method to evaluate the thymus output function. HNA-1 can increase the thymus output function, but there is a certain correlation between TREC, initial CD4 count and thymus histopathology Relationship, especially in the late infection.