健脾补肾解毒法对猴艾滋病的干预效果及猴模型进展因素的研究

发布时间：2018-02-28 06:24

本文关键词： 猴免疫缺陷病毒中医证候药物疗法长期不进展型　出处：《广州中医药大学》2015年博士论文　论文类型：学位论文

【摘要】：目的：评价健脾补肾解毒法中药对猴艾滋病的干预效果；探讨猴艾滋病模型不同类型的进展风险影响因素；探索猴艾滋病模型长期不进展型的内在病理机制；方法：第一章健脾补肾解毒法中药治疗猴艾滋病模型的效果选用同期感染的、基线状态相当的8只艾滋病恒河猴,随机分为两组,分别进行连续8周的中药和抗病毒药治疗,定期采血和采集腹股沟浅表淋巴结,实验结束进行胃肠道粘膜的标本采集。检测T细胞纯真亚群、记忆亚群、功能亚群,肠淋巴归巢受体等免疫指标；血浆病毒载量及组织中SIVRNA和SIVDNA等病毒学指标；以及中医证候指标和病理学结构的检测,分析比较了中药组和抗病毒药组在治疗过程中各指标的差异。第二章猴艾滋病模型不同类型的进展风险影响因素探讨挑选符合长期不进展型(LTNP)、一般进展型(NP)、快速进展型(RP)特点的三种类型恒河猴各10只,与同期5只健康恒河猴对比,分析感染前后不同类型间T细胞亚群、中医证候指标及淋巴结病理的不同之处。找出不同类型进展风险的影响因素,并建立判别方程对其风险进行预测。第三章探索猴艾滋病模型长期不进展型的内在病理机制选入长期不进展型艾滋病恒河猴4只,同期进入试验的3只正常猴(Normal)和4只一般进展型(NP) SIV猴,作为对照,连续观察6个月,观察期间三组均不进行任何干预。定期采血和采集腹股沟浅表淋巴结,实验结束进行胃肠道粘膜的标本采集。检测T细胞纯真亚群、记忆亚群、功能亚群,肠淋巴归巢受体等免疫指标；血浆病毒载量及组织中SIVRNA和SIVDNA等病毒学指标；以及中医证候指标和病理学结构的检测,分析比较三组各指标的差异。结果：第一章健脾补肾解毒法中药治疗猴艾滋病模型的效果健脾补肾解毒法中药治疗虽然不能像HAART一样明显降低血浆和细胞内的SIVDNA/RNA,但可以升高外周血的CD4。该亚群主要为CD4纯真细胞；同时表达CD28上调、CCR5下调、β7+及CCR9明显增加。胃肠道粘膜内β7+及CCR9的表达却明显降低。这一改变,可能与MAVS蛋白下降存在某些关联。此外在一定程度上延缓了脾虚的出现和发展,相比HAART组,淋巴结病理结构保存更完整,均显示了健脾补肾解毒法中药有相当的临床价值。第二章猴艾滋病模型不同类型的进展风险影响因素探讨1各组动物血浆病毒载量均在感染后10-14天达到高峰,平台期出现分化。2与Normal相比,RP型的WBC明显偏高,CD4和LYM比例明显偏低；而LTNP型的LYM数量偏高；3与其它类型相比,RP型的β2-MG显著升高,T3显著降低。4 RP型在平台期出现淋巴结缩小,生发中心明显缩小的退变型,区别于其它各型。感染前WBC和LYM比例进入判别方程,平台期为T4和平台期LoglORNA进入判别方程,对所建立的判别函数的一致率进行检验,感染前和平台期理论判别与实际资料的总吻合率分别为57.1%和91.2%。第三章探索猴艾滋病模型长期不进展型的内在病理机制LTNP组的血浆病毒载量情况、CD4数量、比例和记忆亚群、功能亚群均符合我们的预期,指标大多介于NP组和正常组之间,此外我们发现LTNP组淋巴结中CCR5低表达,趋化于肠粘膜效应部位因子的T细胞更高、且更接近于正常组。这可能是LTNP的重要特征、甚至参与了LTNP的形成。而作为病毒核酸受体TLR、RLR家族共同接头信号蛋白的MAVS在LTNP明显低于NP组,可能使得LTNP淋巴结中炎性细胞因子较低,从而降低了淋巴细胞的活化,进而延缓了疾病进程,使得LTNP猴天然感染SIV病毒载量很低。各指标都表明LTNP处在一种相对稳定状态,而非普通SIV感染处于的活化过度状态。结论：1 该方药可以升高CD4纯真细胞；同时表达CD28上调、CCR5下调、β7及CCR9明显增加,而且淋巴结病理结构保存更完整,有一定的临床价值和应用前景。2 LTNP、NP和RP三种进展类型在平台期有明显差异,其中RP型有更明显的脾肾俱虚证候,提示脾肾俱虚有可能是艾滋病快速进展的关键因素。另外,感染前的WBC和LYM比例可作为评价不同进展类型的一个参考,而平台期的LoglORNA和T4可用于预测不同类型进展的风险。3 LTNP各方面指标介于Normal和NP之间,甚至更偏向于Normal。淋巴结中CCR5低表达,趋化于肠粘膜效应部位因子的T细胞更高、且更接近于正常组,而作为病毒核酸受体TLR、RLR家族共同接头信号蛋白的MAVS在LTNP明显低,可能使得LTNP淋巴结中炎性细胞因子较低,降低了淋巴细胞的活化及疾病进程。这些都可能参与了LTNP的形成,使得LTNP猴天然感染SIV很低。
[Abstract]:Objective: To evaluate the intervention effects of Jianpi Bushen herbs on detoxification of simian AIDS; risk factors in the impact of different types of monkey model of AIDS; explore the internal pathogenesis of simian AIDS type model of long-term development; methods: the period of the first chapter of the infection effect of spleen and kidney detoxification method of Chinese medicine treatment of AIDS monkey model, a baseline the 8 Ganges RIver AIDS monkeys were randomly divided into two groups, respectively, Chinese medicine and antiviral therapy for 8 weeks, and regular blood collection of inguinal superficial lymph node, the end of the experiment specimens of the gastrointestinal mucous membrane. The detection of T cell subsets in pure, memory subsets, functional subsets of intestinal lymphatic homing receptor immune index; plasma viral load of SIVRNA and SIVDNA and the amount of tissue such as virology index; and TCM syndrome index and pathological structure of the detection, analysis and comparison of Chinese Medicine The difference of each index group and antiviral group in the treatment process. The second chapter simian AIDS model factors in the risk of the effects of different types of selected in line with the long term nonprogressors type (LTNP), the general progress of type (NP), rapidly progressive (RP) characteristics of Ganges RIver three types of monkey by 10, and 5 in the same period healthy Ganges RIver monkey comparative analysis before and after infection among different types of T cell subsets, different TCM syndrome index and lymph node pathology. To find out the influencing factors of different types of risk, and establish the discriminant equation to predict the risk. The third chapter explores the intrinsic mechanism of pathological model of simian AIDS type selected for long-term progress the long-term progress of AIDS in Ganges RIver 4 monkeys, 3 normal monkeys into the test period (Normal) and 4 (NP) in general SIV monkey, as control, continuous observation for 6 months, during the observation period, three groups were regularly without any intervention. Blood collection and inguinal superficial lymph node, the end of the experiment were collected of the gastrointestinal mucosa. Detection of T cell subsets in pure, memory subsets, functional groups, intestinal lymphoid homing receptor immune index; plasma viral load of SIVRNA and SIVDNA and the amount of tissue such as virology index; and TCM syndrome index and pathology the structure of the detection, analysis and comparison of each index in three groups. Results: the effect of traditional Chinese medicine of invigorating the spleen and kidney detoxification method of the first chapter of invigorating the spleen and kidney detoxification method of traditional Chinese medicine in the treatment of AIDS treatment although not monkey model like HAART significantly decreased the plasma and intracellular SIVDNA/RNA, but can increase the peripheral blood CD4. of the main CD4 cell subsets of innocence at the same time; increase the expression of CD28, CCR5 and CCR9 reduced, beta 7+ increased significantly. The expression of 7+ beta and CCR9 in gastrointestinal mucosa decreased obviously. This change, there may be some close down and MAVS protein In addition to delay the spleen. The emergence and development to a certain extent, compared with HAART group, lymph node pathology showed more complete preservation, Jianpi Bushen Jiedu method has clinical value. The second chapter is the factors in the risk of simian AIDS model the impact of different types of the discussion in 1 groups of animal plasma viral load in the peak at 10-14 days after infection, the platform differentiation.2 compared with Normal, RP WBC and LYM CD4 were high, the proportion is significantly lower; and the LTNP type LYM high number; 3 compared with other types, RP type beta 2-MG increased significantly, T3 decreased significantly.4 RP type lymph nodes on the platform period of degeneration of the germinal center was significantly reduced, different from other type. WBC and LYM before infection proportion into the discriminant equation. Platform for T4 and LoglORNA platform into the discriminant equation. The discriminant function of the consistent rate Test before infection and the platform theory is consistent with the actual data of the total discrimination rate was LTNP internal pathogenesis group plasma virus 57.1% and 91.2%. the third chapter explore the monkey model of AIDS long-term progressive load, CD4 number, proportion and memory subsets, subsets are in line with our expectations, between the index mostly between NP group and normal group, in addition we found LTNP lymph node in CCR5 low expression of chemotactic factor on intestinal mucosal effector T cells more and more close to the normal group. This may be an important feature of LTNP, and even participate in the formation of LTNP. As the viral nucleic acid receptor TLR, RLR family common joint signal protein MAVS in LTNP was significantly lower than that of NP group, so LTNP lymph node inflammatory cytokines is low, thereby reducing the lymphocyte activation, and thus delay the progress of the disease, the natural LTNP monkey SIV virus infection The load is very low. The index shows that LTNP in a relatively stable state, rather than the common SIV infection is the excessive activation state. Conclusion: the recipe can increase 1 pure CD4 cells; while CD28 expression increased, CCR5 reduced, beta 7 and CCR9 increased significantly, and the lymph node pathological structure more complete preservation that has clinical value and application prospect of.2 LTNP, there are obvious differences in the platform NP and RP three in type, the RP type has more obvious inherent deficiency syndromes of spleen and kidney, spleen and kidney are all hollow tips may be the key factor in the rapid HIV infection. In addition, before the WBC and LYM ratio can be used as a reference to the evaluation of different types of progress, while the platform of LoglORNA and T4 can be used to predict the progress of different types of risk indexes of LTNP.3 between Normal and NP, and even more biased in favor of CCR5 Normal. in lymph nodes of low expression of chemoattractant in the intestinal mucosa. Should part factor in T cell is higher, and more close to the normal group, as the viral nucleic acid receptor TLR, RLR family protein MAVS connector signal in LTNP was significantly lower, may make LTNP lymph node inflammatory cytokines in low reduced lymphocyte activation and progression of the disease. These may be involved in LTNP the formation of the natural infection of SIV LTNP monkey is very low.

【学位授予单位】：广州中医药大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R285.5;R-332

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