基于Ins/PI3K/Akt通路探讨藏药宽筋藤“清除隆热”防治AD的物质基础和作用机制

Alzheimer’s disease(AD), also known as type 3 diabetes, has become a most challenging health concern on the aging society, and there are no effective methods and drugs on its treatment at present. To improve the insulin pathway sensitivity as targets for effective components, is a new breakthrough on AD treatment. Tibetan medicine has a unique curative effect on AD. Tibetan medicine thinks that much more heat obstructed in Long is the core mechanism on the occuring of diabetes, which damages white vein and can induces dementia. Tibetan medicine Caulis Tinosporae, also named as Le Zhe, can clear Long heat, detoxicate, calm the nerves, and is conventionally used in aging disease and AD. In our previous research, Caulis Tinosporae has good effects on AD model rats, and can reduce insulin resistance of diabetic rats. But the research of Caulis Tinosporae is limited to a small amount of medicinal herbs identification, and the efficacy and material base study of Caulis Tinosporae is still very little. In this study, a “Long heat into the brain” AD model combined disease will be proposed to establish with high sugar and high fat diet + STZ intraperitoneal injection + Aβ25-35 hippocampus injection. The effective parts of Caulis Tinosporae will be determined from efficacy index such as ethology, Aβ and p-Tau deposition, hippocampus morphology. The active ingredients of Caulis Tinosporae will be separated and identificated by using reverse phase chromatography, preparative chromatography, molecular imprinting, fingerprint technology, and combined with the classical AD cell model effects. The molecular mechanisms of Caulis Tinosporae on AD will be focus on the Ins/PI3K/Akt pathway. With the active ingredient group or active ingredients monomer as testing index, the quality standards of Caulis Tinosporae medicinal materials on AD will be established. This study will provide an important experimental evidence for the discovery of AD new drugs research, help to explain the Tibetan medicine connotation of “Long heat into the brain” and Caulis Tinosporae “clearing Long heat” to prevent and treat AD, and lay the foundation for the comprehensive development and utilization of Tibetan medicinal materials of Caulis Tinosporae.

阿尔茨海默病（AD）又称3型糖尿病，现有药物疗效不理想。以提高胰岛素通路敏感性为靶点寻找有效成分，是AD治疗新突破点。藏医认为，糖尿病隆热偏盛，入脑损伤白脉，发为痴呆。宽筋藤又名勒哲，在藏族地区习用于治疗衰老病和AD。本课题组前期实验发现宽筋藤有良好的抗AD作用，且可减轻糖尿病大鼠胰岛素抵抗。目前宽筋藤仅限于少量的药材鉴别研究，药效及物质基础研究甚少。本项目拟采用高糖高脂饮食+STZ+Aβ25-35，建立“隆热入脑”AD病证结合动物模型，综合行为学、Aβ和p-Tau沉积、海马形态学等药效，确定宽筋藤有效部位。采用反相色谱、制备色谱、分子印迹、指纹图谱等技术，结合经典AD细胞模型，分离并明晰宽筋藤活性成分，从Ins/PI3K/Akt通路深入研究其作用机制，建立宽筋藤质量标准。可为藏医“隆热入脑”致AD和宽筋藤“清除隆热”治疗AD提供实验依据，对发现AD新药、促进宽筋藤的综合开发有重要意义。

宽筋藤在藏族地区习用于治疗衰老病和AD，但目前宽筋藤防治AD药理作用机制及物质基础研究甚少。本项目采用高糖高脂饮食+STZ+Aβ25-35，建立“隆热入脑”AD病证结合动物模型，综合行为学、Aβ和p-Tau沉积、海马形态学等药效，明确了宽筋藤有效部位。采用硅校柱、反相色谱、制备色谱等技术，分离得到宽筋藤单体化合物48个，其中新化合物3个；结合经典AD细胞模型，明确宽筋藤活性成分为紫丁香苷、Tinosinenside A等；从Ins/PI3K/Akt通路和NFκB/NLRP3/Caspase1通路深入研究有效部位和有效成分的作用机制；采用HPLC法建立宽筋藤药材指纹图谱。本研究可为藏医“隆热入脑”致AD和宽筋藤“清除隆热”治疗AD提供实验依据，对发现AD新药、促进宽筋藤的综合开发有重要意义。