中药复方提取物HNA-1对SIV慢性感染中国恒河猴胸腺输出功能的影响

Effect of Chinese Herbal Extract HNA-1 on the Thymic Output Function in Simian Immunodeficiency Virus Chronically Infected Chinese Rhesus Macaques

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 observe the effect of Chinese herbal extract Hu Nan A-1（ HNA-1） on the thymic output function in Simian immunodeficiency virus（ SIV） chronically infected rhesus macaques. Methods Eight Chinese rhesus macaques had been infected by SIVmac239 for 16 to 21 months,and then they were randomly divided into the treatment group and the control group,4 in each group. Monkeys in the treatment group were administered with HNA-1 by gastrogavage,once daily for 2 successive months,while those in the control group were administered with equal volume of normal saline by gastrogavage,once daily for 2 successive months. The general condition and body weight of monkeys were observed. Plasma viral loads were detected using real-time fluorescent quantitative PCR assay. CD4 percentages and counts,as well as naive CD subsets were detected using flow cytometry. T-cell receptor excision circles（ TREC） were detected using real-time fluores-cent quantitative PCR assay. The thymus tissue was pathologically observed using routine HE staining. The correlation between lesions of the thymus tissue,CD4 counts,naive CD counts,and TREC were analyzed. Results There was no statistical difference in body weight,viral loads,absolute CD ratios between the two groups after treatment（P 〉0. 05）. The altered TREC multiple showed an obvious decreasing tendency in the control group,while it showed an increasing tendency in the treatment group（P〈 0. 05）. In both groups,destroyed structures of the thymus tissue could be seen,filled with pink unstructured material. Increased connective tissues,lowered connective cell density,and confused arrangement could also be seen in the two groups,with no obvious difference. TREC contents were positively correlated with naive CD4 counts after removing extremum（r = 0. 926,P = 0. 001）. Naive CD4 counts were positively correlated with CD4 counts（r = 0. 961,P = 0. 005）. Conclusions TREC content determination,as a marker of newly thymic emigrants,could be taken as a testing method for evaluating the thymic output function. Besides,HNA-1 treatment increased the thymic output significantly in SIV chronically infected monkeys. Correlation existed among TREC contents,naive CD4 counts,and pathologies of thymus tissues,especially in late infection stage.