川芎干预糖尿病肾病的药效物质基础及作用机制

Diabetic nephropathy (DN) is the leading cause of chronic kidney disease in China. ‘Promoting blood circulation and removing blood stasis’ and ‘inhibiting oxidative stress and inflammatory response’ are the predominant approaches for the clinical therapy of DN in traditional Chinese medicines and western medicines, respectively. Chuanxiong Rhizoma is a traditional Chinese medicine possessing the function of ‘promoting blood circulation and removing blood stasis’, and demonstrated a satisfying therapeutic effect against DN. Our previous studies displayed that Chuanxiong Rhizoma displayed potent dual inhibitory effects against oxidative stress and inflammatory response, which was consistent with the therapeutic approaches of traditional Chinese medicines and western medicines. It contained numerous bioactive substances with diverse structures and different mechanism of action. Lots of bioactive substances have not been verified. Phthalide possess potent dual inhibition against oxidative stress and inflammatory response, and demonstrated potential as new therapeutic drugs against DN. Based on these observations, the work will be carried out as follows. (i) Bio-guided isolation and structure elucidation of substances with inhibitions on oxidative stress and inflammatory response, and establishing the relationships between biological activities and chemical substances. (ii) Based on the key oxidative stress- and inflammation- regulating factors involving the DN pathological process, investigating the molecular mechanism of different types of bioactive substances, and discussing their synergic effect of different structures. (iii) Using the rational cell and animal models, evaluating the therapeutic effect of bioactive substances (e.g. phthalides) against DN. The results of this project will illustrate its bioactive substances of Chuanxiong Rhizoma, give the chemical and biological evidences for its traditional applications, provide a basis for research and development of new DN drugs, and are significant for finding DN therapeutic agents from traditional Chinese medicines.

糖尿病肾病(DN)是我国慢性肾病的首要病因。“活血化瘀”与“抑制氧化应激和炎症反应”是中西医治疗DN的主要策略。川芎为活血化瘀中药，临床治疗DN疗效确切。申请人前期研究发现：川芎双重抑制氧化应激和炎症反应作用显著，与中西医治疗DN的原则相符；活性成分结构多样、数目繁多、机制迥异，仍有大量强活性成分尚未明确；苯酞类成分有强氧化应激和炎症反应双重抑制作用，有成为新型抗DN药物的潜力。本项目拟：（1）开展川芎氧化应激和炎症反应双重抑制活性成分追踪分离和构效关系分析，阐明成分与活性的相关性；（2）针对DN病理进程中关键的氧化应激和炎症反应调控机理，揭示不同结构类型强活性成分的作用机制和协同作用；（3）开展活性成分（尤其苯酞类）干预DN的体内外有效性研究。研究结果将揭示川芎药效物质基础，完善其中药理论和传统应用的化学、生物学与药理学依据，为新型抗DN药物研发奠定基础，对抗DN中药基础研究有重要意义。

糖尿病肾病(Diabetic nephropathy,DN)是我国慢性肾病的首要病因。“活血化瘀”与“抑制氧化应激和炎症反应”是中西医治疗DN的主要策略。川芎为活血化瘀中药，临床治疗DN疗效确切。本项目以川芎为研究对象，围绕Nrf2、NF-kB和NLRP3炎症小体三个调控氧化应激和炎症反应关键通路，针对糖尿病肾病这一重大疾病，开展研究工作，包括：（1）开展了川芎中具有氧化应激和炎症反应抑制活性成分的分离制备和结构鉴定，分离获得55个化学成分（新化合物3个），包括简单苯酞、苯酞二聚体、芳香化合物、糖类等化学成分类型，评价了分离获得成分的氧化应激和炎症反应抑制作用，发现苯酞是主要活性成分类型。（2）富集制备了苯酞类活性组分D70，发现其能够改善DN小鼠血糖的变化，缓解肾小球的肥大和肾脏纤维化，有效地缓解小鼠DN病变，其肾脏保护作用与调控Nrf2、NF-kB、Vimentin、α-SMA抑制肾脏组织氧化应激和炎症反应相关。（3）完成了强活性苯酞类单体化合物体内抑制炎症有效性评价，四个苯酞类成分能够有效地改善脂多糖诱发的组织炎症反应。（4）发现川芎中糖类成分通过抑制α-糖苷酶活性降低小鼠血糖并改善DN病变进程，无抑制氧化应激和炎症反应作用。（5）开展了生物碱干预DN研究，发现生物碱LH通过调控NF-kB信号通路和NLRP3炎症小体发挥抑制炎症反应作用，进而改善高血糖诱发的小鼠肾脏肥大、肾小球系膜基质的扩张和肾纤维化，具有干预DN作用。（6）构建了中药活性成分干预氧化应激和炎症反应相关疾病研究技术体系，为深入开展中药干预DN药效物质与分子机制研究提供了技术支撑。上述研究结果揭示了川芎干预DN的药效物质基础，阐明了传统医学使用川芎治疗DN的科学性，确证了川芎活性成分成为DN干预药物的潜力，为治疗DN新药研发奠定基础。