健脾补肾解毒法对艾滋病干预作用的胃肠道免疫及病理研究

Gastrointestinal barrier damage and leading to excessive immune activation is a key factor in HIV patients progress to AIDS, antiviral therapy can reduce HIV viral load and increase CD4 cell count of the peripheral blood, but no obvious improvement of gastrointestinal function.BOTA FOR AID?Capsule（BFA），based on principles of invigorating spleen and kidney detoxification method,can significantly reduce fatigue、bad appetite、abdominal distension and other gastrointestinal symptoms，make the weight increased and improve quality of life among people infected with HIV.Furthermore,the experimental study shows that BFA can improve pathological changes of the lymph nodes in SIV infected monkeys significantly than that in antiviral therapy group, speculated that the the invigorating spleen and kidney detoxification medicine may have certain improvement effect on the gastrointestinal immune function and pathology of HIV patients. This project intends to adopt the monkey AIDS model from the perspective of gastrointestinal mucosal immune function and pathology of the invigorating spleen and kidney detoxification medicine for HIV/AIDS intervention effect,use flow cytometry,quantitative PCR analysis, radioimmunity method,immunohistochemical method comprehensively expound immune function and the characteristic and regularity of pathological changes of gastrointestinal mucosa,mesenteric lymph nodes and Peyer's knot,superficial lymph nodes and peripheral blood in SIV infected monkeys.It will prompt new ideas about deep understanding pathological mechanism for AIDS and improve TCM treatment effect on AIDS, promote AIDS prevention and control work in our country.

胃肠道屏障破坏而导致免疫过度活化是HIV感染者进展至AIDS的关键因素，抗病毒疗法能降低外周血HIV病毒载量、升高CD4细胞数，但对胃肠道功能无明显改善作用。以健脾补肾解毒法为原则的艾可清复方能明显减轻HIV感染者乏力、纳差、腹胀等胃肠道症状，并使患者体重增加生存质量改善；且实验研究显示，艾可清对SIV感染猴的淋巴结病理改善明显优于抗病毒药组，因此推测健脾补肾解毒法可能对HIV感染者的胃肠道免疫功能及病理有一定改善作用。本项目拟采用猴艾滋病模型从胃肠道粘膜免疫功能及病理角度探讨健脾补肾解毒法对艾滋病的干预效果，利用流式细胞术、定量PCR、放射免疫法、免疫组化法等技术较全面阐明艾滋病胃肠道粘膜、肠系膜淋巴结、Peyer's结、浅表淋巴结及外周血的免疫病理变化特点及规律，揭示肠道免疫与外周免疫的关系，为AIDS病理机制的更深刻认识及中医治疗AIDS疗效的提高提示新思路，推动我国艾滋病防治工作。

本研究从外周血、浅表淋巴结及胃肠道等不同组织角度，以抗病毒药物为对照，评价了健脾补肾解毒法方药干预猴艾滋病的效果，并进一步探索了长期不进展（LTNP）猴艾滋病模型相应组织和免疫指标的免疫病理变化规律。项目运用流式细胞术、定量PCR、放射免疫法、免疫组化法等技术检测了各组织中T细胞亚群（CD3/4/8，CD28，CD45RA），趋化因子受体CCR5水平，线粒体抗病毒信号蛋白MAVS，肠淋巴归巢指标，血清内毒素LPS，血浆及组织中病毒载量，中医证候指标等的变化，并进行了病理观察。.研究结果：以健脾补肾解毒法方药虽不能像HAART一样显著降低血浆和细胞内的病毒载量，但有更好的免疫调节功能：升高外周血CD4纯真细胞，同时表达CD28上调、CCR5下调、β7及CCR9明显增加、MAVS显著减少；恰恰与作为免疫治疗活指南的LTNP型中CCR5低表达、CCR9+CD3+高表达、MAVS低表达相一致，这是该方药的可贵之处。作为病毒核酸受体TLR、RLR家族共同接头信号蛋白的MAVS低表达，可能使得淋巴结中炎性细胞因子较低，从而影响了T细胞归巢特性的形成、降低了淋巴细胞的活化及疾病进程，其可能是SIV感染中天然免疫和细胞免疫衔接转换的桥梁，关于MAVS的通路机制有必要增大样本量进一步深入研究。此外健脾补肾解毒法方药在一定程度上延缓了脾虚的出现和发展，对胸腺、脾脏、浅表淋巴结病理结构保存更完整，结合中药组实验动物的临床症状表现及存活情况均优于抗病毒药组，提示我们在治疗艾滋病时，健脾补肾解毒法方药的免疫调节作用与抗病毒药的病毒抑制作用同等重要，应该优势互补，该方药有相当的临床价值和应用前景。.本研究部分阐明了健脾补肾解毒法对艾滋病干预作用的胃肠道免疫及病理，发现了中药对胃肠道免疫和病理的作用特点，为形成艾滋病的中医药治疗方案提供了科学依据，为AIDS病理机制的更深刻认识及中医治疗AIDS疗效的提高提示新思路，对推动我国艾滋病防治工作有着现实意义。