虫草益肾方通过腺苷信号通路延缓肾间质纤维化的机制研究  

Study on mechanism of Chongcaoyishen Formula in delaying TIF by adenosine signaling pathway

作　　者：范桢亮 宋业旭[2] 贠捷[2,3] 宋立群[2,4] FAN Zhen-liang;SONG Ye-xu;YUN Jie;SONG Li-qun(The First Affiliated Hospital of Zhejiang Chinese Medical University(Zhejiang Provincial Hospital of Chinese Medicine),Hangzhou 310006,China;Heilongjiang University of Chinese Medicine,Harbin 150040,China;First Affiliated Hospital,Heilongjiang University of Chinese Medicine,Harbin 150040,China)

机构地区：[1]浙江中医药大学附属第一医院(浙江省中医院),杭州310006 [2]黑龙江中医药大学,哈尔滨150040 [3]黑龙江中医药大学附属第一医院,哈尔滨150040 [4]黑龙江中医药大学附属第一医院肾内科,哈尔滨150040

出　　处：《中华中医药杂志》2020年第9期4661-4665,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然青年科学基金项目(No.81804088);黑龙江中医药大学研究生创新科研项目(No.2018yjscx007);教育部“春晖计划”合作科研项目(No.Z2008-1-15020)。

摘　　要：目的:探究虫草益肾方调控腺苷信号通路抗肾纤维化(TIF)的作用机制。方法:选用雄性SD大鼠并随机分成空白组、假手术组、模型组、虫草益肾方组、腺苷组、茶碱组,每组6只。制备UUO模型后第1天起,虫草益肾方组给予虫草益肾方(0.00657g·g^-1·d^-1)灌胃,腺苷组给予腺苷(0.0015mg·g^-1·d^-1)、茶碱组给予茶碱溶液(0.01mg·g^-1·d^-1)腹腔注射,14d后采集大鼠血清与肾组织分别进行组织病理学(HE&Masson染色)、血清炎症因子(TNF-α、IL^-1β/6/10/12)、免疫组化(α-SMA、COL-Ⅲ)、Western Blot(α-SMA、COL-Ⅲ、A2AAR、A2BAR)与RT-PCR(α-SMA、COL-Ⅲ、A2AAR、A2BAR)检测。结果:模型组与腺苷组大鼠血清中炎症因子均显著升高,相比于空白组与假手术组出现了显著的TIF(P<0.05)。而虫草益肾方与茶碱均可明显抑制大鼠炎症反应与肾组织纤维化,并且虫草益肾方疗效更优(P<0.05)。此外,虫草益肾方可以上调肾组织中A2AAR水平并抑制A2BAR表达,对腺苷信号通路产生特殊的调控作用。结论:虫草益肾方可能通过调控腺苷信号通路发挥其延缓TIF的作用。Objective:To explore the mechanism of adenosine signaling pathway mediated by Chongcaoyishen Formula against renal fibrosis(TIF).Methods:Male SD rats were randomly divided into Blank Group,Sham Group,UUO Group,Chongcaoyishen Formula Group,Adenosine Group and Theophylline Group,with 6 rats in each group.On the first day after UUO model was prepared,the Chongcaoyishen Formula Group was given intraperitoneal injection(0.00657 g·g^-1·d^-1),Adenosine Group was given adenosine(0.0015 mg·g^-1·d^-1),and Theophylline Solution(0.01 mg·g^-1·d^-1)was given intraperitoneal injection.After 14 days,serum and kidney tissues were collected for histopathology(HE&Masson staining),serum inflammatory factors(TNF-α,IL^-1β/6/10/12),immunohistochemistry(α-SMA,COL-Ⅲ,A2 AAR,A2 BAR),Western Blot(α-SMA,COL-Ⅲ,A2 AAR,A2 BAR),and RT-PCR(α-SMA,COL-Ⅲ,A2 AAR,A2 BAR).Results:Inflammatory cytokines were significantly increased in the UUO Group and Adenosine Group,and TIF was significantly higher than that in normal renal tissue(P<0.05).Both the Chongcaoyishen Formula and theophylline could significantly inhibit the inflammatory response and renal tissue fibrosis in rats,and the Chongcaoyishen Formula had a better effect(P<0.05).In addition,Chongcaoyishen Formula can up-regulate the A2 AAR level and inhibit the expression of A2 BAR in renal tissues,thus exerting a special regulatory effect on adenosine signaling pathway.Conclusion:Chongcaoyishen Formula may play the role of delaying TIF by regulating adenosine signaling pathway.

关 键 词：虫草益肾方 肾小管间质纤维化 慢性肾脏病 腺苷信号通路 单侧输尿管梗阻 

分 类 号：R285.5[医药卫生—中药学]