犀角地黄汤调控巨噬细胞有氧糖酵解干预脓毒症的机制研究

Sepsis is an usual critical ill in clinical medicine, with a high incidence rate in all specialties, especially in the department of Intensive Care Unit (ICU).The mortality of septic shock is up to 70%, yet now we are lack of effective drugs. A long-term clinical practice by our research group authenticate that Xijiao Dihuang decoction (XJDHT) can significantly improve the prognosis of Sepsis, shows a good application prospect. Combined with previous research results and literature survey, we consider that the new mechanism of treating Sepsis by XJDHT is probably down regulation the aerobic glycolysis in macrophages. This subject clarify that XJDHT can act on aerobic glycolysis of macrophage, depress the excessive release of inflammatory mediators, allay the accumulation of lactic acid, through the establishment of in-vivo and in-vitro models. The key target is probably relevant to the depress of the signaling pathway “TLR4/HIF1α/PKM2”.Our reseach clarify the action mechanism of treating Sepsis by XJDHT, from a new perspective, reveal the essence of treating sepsis by traditional Chinese medine, provide the reliable experimental basis for developing new drugs, possess considerable clinical application value.

脓毒症是临床常见的危急重症，在各专科特别是重症医学科有较高发生率，脓毒性休克死亡率高达70%，但目前仍缺乏行之有效的治疗药物。课题组长期临床实践证实犀角地黄汤（XJDHT）可有效改善脓毒症患者预后，显示出了良好的应用前景。结合前期研究成果及文献调查，我们认为下调巨噬细胞有氧糖酵解代谢水平，有可能是犀角地黄汤治疗脓毒症的新机制。本项目通过构建体内外模型，结合现代分子生物学手段，基于巨噬细胞有氧糖酵解代谢角度，诠释XJDHT有效降低炎性介质的过度释放和乳酸的过度蓄积，其关键靶点有可能与抑制“TLR4/HIF1α/PKM2”信号通路有关。本研究从新的角度更加深入阐明XJDHT治疗脓毒症的作用机制，揭示了传统中医药治疗脓毒症的现代科学本质，为开发针对巨噬细胞有氧糖酵解的新药提供可靠的实验依据，具有可观的临床应用价值。

有氧糖酵解是加重脓毒症进展的关键因素。犀角地黄汤已被证明对脓毒症具有良好的治疗作用。前期的临床研究表明，犀角地黄汤能够改善脓毒症的生存率。在本研究中，我们研究了犀角地黄汤对脓毒症时有氧糖酵解的影响。研究中，我们将脓毒症大鼠模型随机分为5组，分别为对照组，模型组，TAK-242（TLR4 抑制剂）组，犀角地黄汤（25g/kg）组和犀角地黄汤（12.5g/kg）组。结果发现，与对照组相比，模型组细胞因子IL-1β、IL6的含量增加，而犀角地黄汤显著降低了IL-1β、IL-6等细胞因子表达。同时，模型组TLR4、HIF-1α、PKM2等关键因子表达较空白组显著增加，犀角地黄汤可显著降低TLR4，HIF-1α和PKM2的表达。本研究共设立五个组，包括对照组，LPS组，siTLR4组，犀角地黄汤（4mg/mL）组和犀角地黄汤（2mg/mL）组体外实验。 LPS刺激后，模型组中的IL-1β和IL-6显着升高，而犀角地黄汤降低了细胞因子的表达。通过刺激LPS，TLR4，HIF1α和PKM2的蛋白表达显着增加，而犀角地黄汤下调了信号通路中关键分子的表达。这项研究表明犀角地黄汤能够通过下调TLR4/HIF-1α/PKM2信号通路抑制有氧糖酵解来改善脓毒症的预后。