本课题前期发现新疆鼠尾草总酚酸可改善糖尿病肾病大鼠症状，作用机制研究表明：其可减少机体中AGEs的生成，增强血管壁弹性，降低血管通透性；降低醛糖还原酶活性，减少细胞内山梨醇形成，改善细胞内渗透压；可抑制细胞内二酰甘油（DAG）的合成，下调TGF-β1和VEGF的表达，推测其通过抑制PKC的活性而发挥作用。同时，其可下调TNF-α和NO水平，增强血清中SOD活性，降低血清中 MDA 的含量，提示其可通过改善糖尿病肾病大鼠的氧化应激及炎症反应，降低肾脏组织的损伤，从而延缓糖尿病肾病的发展。本课题拟用工业级HPLC制备色谱从有效部位中大量制备前期鉴定的11种单体酚酸成分，并测试其改善STZ所致高血糖大鼠糖尿病肾病作用，找到其活性成分；利用体内外糖尿病肾病模型，探讨新疆鼠尾草中酚酸类成分协同调控PKC网络、抗氧化应激和炎症反应的细胞内蛋白调控机制，为糖尿病肾病的治疗提供理论依据和新药先导化合物。

In our initial studies, we have found total phenolic acid in Salvia desert SHANG can relieve diabetic nephropathy in rats. Regarding mechanisms, our studies have revealed that they can prevent production of AGEs in organism, strengthen elasticity of blood vessel wall, and reduce permeability of blood vessels. And, they have abilities of decreasing activities of aldose reductase, prevent formation of sorbitol, and improve conditions of osmotic pressure in cells. They can inhibit synthesis of DAG in cells, down-regulate expression of TGF-β1 and VEGF, which are supposed to act through inhibit activities of PKC. Also, they are able to down-regulate expression of TNF-α and NO, enhance activities of SOD, decrease levels of MDA in serum, which indicates they can mitigate damage of kidney tissues, and hence delay development of diabetic nephropathy through improving conditions of oxidative stress and inflammatory reactions in rat model of diabetic nephropathy. In this project, we are going to perform large-scale preparation of 11 phenolic acid compounds we have identified previously in Salvia desert through industrial grade preparative chromatography of HPLC, and find active compounds which can relieve diabetic nephropathy in hyperglycaemia rats, and by using in vivo and in vitro diabetic nephropathy models, with the goal of exploring protein regulation mechanisms of regulation effects of phenolic acids in Salvia desert on PKC network, anti-oxidation stress, and inflammatory reactions, which would eventually provide theoretical evidence and lead drugs for treatment of diabetic nephropathy.

本课题在前期研究中发现新疆鼠尾草总酚酸可改善糖尿病肾病大鼠症状，在此基础上，进一步对新疆鼠尾草中酚酸类化学成分防治糖尿病肾病的作用和机制进行研究。主要研究内容：1、系统分离纯化新疆鼠尾草酚酸类成分，用波谱学方法鉴定出7个单体化合物。分别为：丹参素、丹参素甲酯、原儿茶醛、原儿茶酸、咖啡酸、迷迭香酸、丹酚酸K，其中丹参素、丹参素甲酯、原儿茶醛、原儿茶酸、咖啡酸这5个化合物均首次从新疆鼠尾草中分离得到。2、以总酚酸含量为考察指标，通过单因素考察和正交实验确立新疆鼠尾草总酚酸的最佳提取工艺为：药材加25倍量40%乙醇，回流提取3次，每次1小时，得到新疆鼠尾草药材中总酚酸的含量为62.81mg？g-1；3、腹腔注射链脲佐菌素诱导大鼠成糖尿病肾脏损伤模型，给药灌胃，研究新疆鼠尾草总酚酸提取物、迷迭香酸和丹酚酸K对糖尿病大鼠血糖、血脂、氧化功能、肾脏功能的影响及一些生长因子和细胞因子在糖尿病大鼠血清中的表达，评价新疆鼠尾草对糖尿病大鼠肾脏病变的改善作用，探讨其防治糖尿病肾脏病变的机制。结果：与模型组大鼠比较，新疆鼠尾草丹酚酸K可以改善高糖高脂的糖尿病大鼠的肾脏病变，具体表现在：①降低糖尿病大鼠的血糖水平（P＜0.05）；② 使血清中甘油三酯及低密度脂蛋白的含量显著性降低（P＜0.05）；③ 降低糖尿病大鼠血清中肌酐含量（P＜0.01）及血尿素水平（P＜0.05）；④ 降低血清中谷草转氨酶水平（P＜0.05）；⑤显著升高血清中SOD水平，降低血清中MDA含量（P＜0.01）；⑥降低血清中NO水平，（P＜0.01）；⑦降低血清中的AGEs水平（P＜0.01）；⑧降低血清中的TNF-α含量（P＜0.01）；⑨免疫组化方法检测到各剂量组肾脏组织中TGF-β1和VEGF阳性染色程度均下降，阳性染色面积的积分光密度值均下降（P＜0.01）。与模型组相比，总酚酸提取物及迷迭香酸对糖尿病大鼠空腹血糖、糖化血红蛋白均有降低作用，对糖尿病大鼠，肾脏指数，CREA含量的影响也有统计学意义。本研究为新疆鼠尾草预防和治疗DN提供了更深入的理论依据，在延缓DN的发生和发展具有重要的临床意义，并有助于进一步对新疆鼠尾草开发和利用。已投稿论文1篇，申请1项国家发明专利。培养博士研究生1名, 硕士研究生1名。