滋肾丸方调控胆汁酸代谢改善糖尿病脑病作用机制研究  

Study on the mechanism of regulating bile acid metabolism to improve diabetic encephalopathy by Zishenwan Prescription

作　　者：刘平 杨艮辉 孟凡钰 李英 许梦习 郭虹[2] 张艳军[3] 尹清晟 庄朋伟[1] Ping Liu;Genhui Yang;Fanyu Meng;Ying Li;Mengxi Xu;Hong Guo;Yanjun Zhang;Qingsheng Yin;Pengwei Zhuang(School of Chinese Materia Medica,Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China;Institute of Chinese Medicine of Tianjin University of Traditional Chinese Medicine,Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China;The First Affiliated Hospital of Tianjin University of Traditional Chinese Medicine,National Clinical Medical Research Center of Acupuncture and Moxibustion,Tianjin 300381,China)

机构地区：[1]天津中医药大学中药学院,天津301617 [2]天津中医药大学中医药研究院,天津301617 [3]天津中医药大学第一附属医院、国家中医针灸临床医学研究中心,天津300381

出　　处：《国际中医中药杂志》2024年第7期860-866,共7页International Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(82174112)。

摘　　要：目的考察滋肾丸方对糖尿病脑病小鼠胆汁酸代谢的影响, 探讨其改善糖尿病脑病的作用机制。方法选用雄性C57BL/6J小鼠, 采用高脂饲料喂养联合单次腹腔注射链脲佐菌素(120 mg/kg)复制T2DM小鼠模型, 在高血糖持续刺激8周后采用Morris水迷宫实验筛选糖尿病脑病模型小鼠, 并将造模成功小鼠按随机数字表法分为模型组和滋肾丸方组, 每组12只, 另设12只为对照组。滋肾丸方组小鼠灌胃滋肾丸方粗提取物9.36 g/kg, 对照组和模型组灌胃等体积蒸馏水, 1次/d, 持续8周。采用Morris水迷宫实验检测小鼠认知功能, 采用甲酚紫染色检测海马区颗粒神经元数量, 采用液质联用技术检测血清及粪便胆汁酸含量, 采用荧光定量PCR实验检测肝脏胆汁酸合成酶胆固醇7α-羟化酶(CYP7a1)、甾醇-27-羟化酶(CYP27a1)、法尼醇X受体(FXR)、成纤维生长因子15(FGF15)、成纤维生长因子受体4(FGFR4), 回肠顶端钠依赖性胆汁酸转运蛋白(ABST)mRNA水平。结果与模型组比较, 滋肾丸方组小鼠逃避潜伏期缩短(P<0.05或P<0.01), 首次到达平台时间减少(P<0.01), 穿越平台次数增加(P<0.01), 海马区神经细胞损伤减轻;滋肾丸方组小鼠血清及粪便总胆汁酸含量降低(P<0.05或P<0.01);肝脏中CYP7a1、CYP27a1 mRNA水平增加(P<0.01), FXR、FGF15 mRNA水平降低(P<0.01);回肠ABST mRNA水平降低(P<0.01)。结论滋肾丸方可能调控胆汁酸代谢, 抑制FRX-FGF15/FGFR4信号和ABST表达从而促进新胆汁酸合成和结合型胆汁酸重吸收, 进而发挥改善糖尿病脑病小鼠认知功能的作用。Objective To examine the effects of Zishenwan Prescription on bile acid metabolism in mice with diabetic encephalopathy;To explore its mechanism of improvement of diabetic encephalopathy.Methods Male C57BL/6J mice were used to replicate the mouse model of type 2 diabetes mellitus by using high-fat chow and a single intraperitoneal injection of streptozotocin(120 mg/kg).The mice were screened for diabetic encephalopathy by using the Morris water maze test after 8 weeks of continuous stimulation with hyperglycemia,and were divided into model group and Zishenwan Prescription group according to random number table method,with 12 mice in each group.The mice in the Zishenwan Prescription group were treated with the crude extract of Zishenwan Prescription(9.36 g/kg)by gavage,and the normal group and the model group were treated with the same volume of distilled water once a day for 8 weeks.At the end of the treatment,Morris water maze test was used to investigate the cognitive function of diabetic encephalopathy mice;cresyl violet staining was used to detect the number of granule neurons in the hippocampus;serum and feces were collected to detect the content of bile acids by liquid-liquid coupling;hepatic bile acid synthase CYP7a1 and CYP27a1,farnesol X receptor(FXR),fibroblast growth factor 15(FGF15),fibroblast growth factor receptor 4(FGFR4),and ileocecal apical sodium-dependent bile acid transporter protein(ABST)mRNA levels were detected by using fluorescence quantitative PCR assay.Results Compared with the model group,mice in the Zishenwan Prescription group had shorter evasion latency time(P<0.05 or P<0.01),decreased time to first reach the platform(P<0.01),increased number of times to traverse the platform(P<0.01),and reduced neuronal cell damage in hippocampal area;mice in the Zishenwan Prescription group showed decreased serum and fecal total bile acid content(P<0.05 or P<0.01);the liver CYP7a1 and CYP27a1 mRNA expressions increased(P<0.01),and FXR and FGF15 mRNA expressions decreased(P<0.01);ileal ABST mRNA exp

关 键 词：胆汁酸类 代谢 糖尿病脑病 滋肾丸方 FXR-FGF15/FGFR4信号通路 小鼠 

分 类 号：R285.5[医药卫生—中药学]