万氏牛黄清心丸中朱砂配伍减毒作用的物质基础和机制研究

Cinnabar (HgS) is a famous traditional Chinese Medicine (TCM), which can cause renal injury owing to mercury accumulation in the case of unreasonable use. Wanshi Niuhuang Qingxin Wan (WSNHQXW) is a classic prescription of TCM, which consists of six TCMs, i.e. calculus bovis, cinnabar, rhizoma coptidis, scutellaria, gardenia and radix curcumae. Cinnabar accounts for 10.2% in WSNHQXW. Our study showed that cinnabar could cause renal interstitial inflammation and slight fibrogenesis in rats. Given that all of rhizoma coptidis, scutellaria, gardenia, radix curcumae and calculus bovis have anti-inflammatory effect, we assume that one or several TCM in WSNHQXW could attenuate cinnabar-induced renal interstitial inflammation. In this study, subchronic test of rats will be done, and renal function, pathology and mercury content will be detected to evaluate the renal toxicity of cinnabar and WSNHQXW. To explore which play a role in detoxification, rats will be treated with cinnabar, cinnabar combined with another TCM in WSNHQXW, or WSNHQXW, and the renal toxicity will be compared. On this basis, rats will be further treated with cinnabar combined with active components, and the renal toxicity will be compared with that of cinnabar and cinnabar combined with the corresponding TCM in order to explore the material basis of detoxification. Moreover, renal expression of mediators related to inflammation and fibrosis will be detected to reveal the mechanism of detoxification. The study will be helpful to clarify the scientific connotation of the recipe of WSNHQXW, provide theoretical basis for guidance of its rational clinical use and promote the modernization of TCM.

朱砂(HgS)为著名中药，不科学服用可引起汞蓄积性肾损伤。万氏牛黄清心丸为经典中药方剂，由牛黄、朱砂、黄连、黄芩、栀子、郁金六味药组成，朱砂含量占10.2%。我们的实验研究显示朱砂可引起大鼠肾间质炎症和轻度纤维增生。黄连、黄芩、栀子、郁金和牛黄都具有抗炎作用，因此推测万氏牛黄清心丸中的某(几)味中药对朱砂致肾间质炎症具有配伍减毒作用。本研究拟对大鼠进行亚慢性染毒，通过肾汞含量、肾功能和病理检测，比较朱砂与万氏牛黄清心丸的肾毒性；通过朱砂、朱砂+各单味药、万氏牛黄清心丸的肾毒性比较，探明对朱砂具有减毒作用的单味药；再通过朱砂、朱砂+单味药中的抗炎活性组分、朱砂+单味药的肾毒性比较，探明万氏牛黄清心丸中对朱砂致大鼠肾间质炎症具有减毒作用的物质基础，通过检测肾脏炎症纤维化相关因子表达的改变探索减毒机制。本项目将为解释万氏牛黄清心丸组方的科学性、指导临床合理用药及推进中药现代化建设提供理论依据。

朱砂（HgS）是一种含汞的中药，有肾毒性；万氏牛黄清心丸是一种经典中药方剂，由牛黄、朱砂、黄连、黄芩、栀子、郁金六味药组成。方剂配伍是中医用药的特色之一，合理配伍可起到增效减毒的作用。本课题以万氏牛黄清心丸为研究对象，采用亚慢性肾毒性大鼠模型，对朱砂配伍减毒作用的物质基础及机制进行了研究。研究发现：（1）万氏牛黄清心丸可减少朱砂引起的肾汞蓄积，减轻肾损伤。（2）组方中的黄连、栀子、郁金对朱砂具有减毒作用，单味药的减毒效果不如全方明显；其机制主要通过减少血汞吸收和/或促进尿汞排出，进而减少肾汞蓄积；黄芩可能具有一定的减毒作用；牛黄对朱砂引起的肾汞蓄积拮抗作用不明显。（3）黄连碱和小檗碱是黄连对朱砂具有减毒作用的物质基础，其机制包括：减少血汞吸收，降低肾汞蓄积；下调促炎症细胞因子IL-1β、TNF-α的表达，上调抗炎细胞因子IL-4、IL-10的表达。（4）栀子苷是栀子对朱砂减毒作用的物质基础，其机制为减少血汞吸收，促进尿汞排出，降低肾汞蓄积。本研究完成了课题的预期目标，为解释万氏牛黄清心丸组方的科学性、指导临床合理用药及推进中药现代化建设提供了理论依据。