桂附地黄丸改善自然衰老小鼠肠道脂质吸收功能减退的研究  

Effect of Guifu Dihuang Wan on Intestinal Lipid Absorption in Naturally Aged Mice

作　　者：郭煜晖[1,2,3] 潘冯缘 李梅 唐小涵[1,2] 张长城 童乐 GUO Yuhui;PAN Fengyuan;LI Mei;TANG Xiaohan;ZHANG Changcheng;TONG Le(College of Medicine and Health Sciences,Three Gorges University,Yichang 443002,China;Third-Grade Pharmacological Laboratory on Traditional Chinese Medicine,State Administration of Traditional Chinese Medicine,Three Gorges University,Yichang 443002,China;Hubei Clinical Research Center for Functional Digestive Diseases of Traditional Chinese Medicine and Hospital of Traditional Chinese Medicine of Three Gorges University,Yichang 443005,China;College of Basic Medicine Sciences,Three Gorges University,Yichang 443002,China)

机构地区：[1]三峡大学健康医学院,湖北宜昌443002 [2]国家中医药管理局中药药理科研三级实验室,湖北宜昌443002 [3]湖北省功能性消化系统疾病中医临床医学研究中心/三峡大学中医医院,湖北宜昌443005 [4]三峡大学基础医学院,湖北宜昌443002

出　　处：《中国药学杂志》2024年第16期1513-1521,共9页Chinese Pharmaceutical Journal

基　　金：国家自然科学基金项目资助(81873349);湖北省功能性消化系统疾病中医临床医学研究中心开放基金资助(SXZ202314)。

摘　　要：目的 探究桂附地黄丸对衰老小鼠肠道脂质吸收功能减退的改善作用及可能的作用机制。方法 将26只SPF级雄性11月龄雄性C57BL/6J小鼠饲养至21月龄,随后分为衰老模型组(Model组)和桂附地黄丸给药组(GFDHW,1.17 g·kg^(-1)),给药组小鼠采用饲料给药的方法,持续给药3个月,给药期间自由饮水,Model组小鼠给予正常饲料饲养,期间自由饮水,记录各组小鼠每周的体质量变化,于24月龄时进行处理取材;9只SPF级雄性8月龄雄性C57BL/6J小鼠作为青年对照组(Con组),适应性饲养1周后进行处理取材。取材处理前,各组小鼠禁食24h,禁食结束后各组小鼠灌胃0.2 mL橄榄油,取材处理前30 min,各组小鼠灌胃0.2 mL半固体营养液。小肠推进运动实验检测小鼠小肠运动能力;生化试剂盒检测各组小鼠血清甘油三酯(triacylglycerol,TG)、总胆固醇(total cholesterol,TC)和低密度脂蛋白(low-density lipoprotein,LDL)的含量;透射电镜观察各组肠上皮细胞中脂滴数量变化;钙钴法碱性磷酸酶染色检测各组肠道碱性磷酸酶(intestinal alkaline phosphatase,IAP)活性;苏木素-伊红(hematoxylin eosin,HE)染色观察各组肠上皮形态;免疫组织化学法检测各组小鼠小肠Olfm4、Lgr5、PCNA、Notch1、Hes1以及Villin的表达;免疫印记法(Western blot)检测各组小鼠小肠脂质吸收相关分子蛋白表达水平。结果 与Con组小鼠相比,Model组小鼠体质量下降较快,小肠运动能力下降,血清TG、TC、LDL含量增加,肠上皮细胞中脂滴数量增多,GFDHW组小鼠较衰老小鼠小肠运动能力有所提升,血清TG、TC、LDL含量降低,肠上皮细胞中脂滴数量减少;HE染色Model组小鼠小肠绒毛排列紊乱、断裂,绒毛长度缩短,隐窝深度降低,隐窝数量减少,与衰老小鼠相比,GFDHW组小鼠小肠绒毛排列整齐、连续,绒毛长度增加,隐窝深度升高,隐窝数量增多;免疫组化结果显示,与Con组小鼠相比,Model组小鼠小肠Olfm4、PCNA、Notch1�OBJECTIVE To investigate the ameliorative effect of Guifu Dihuang Wan(GFDHW) on intestinal lipid absorption dysfunction in aging mice and its potential mechanisms.METHODS Twenty-six SPF male C57BL/6J mice aged 11 months were raised to 21 months and divided into an aging model group(Model group) and a GFDHW group(1.17 g·kg~(-1)).The GFDHW group received medication via feed administration for 3 months,with free access to water during the administration period,while the Model group was fed normal feed and had free access to water.Weekly changes in body weight were recorded for each group of mice,and specimens were collected at 24 months of age.Nine SPF male C57BL/6J mice aged 8 months were used as the youth control group(Con group) and were acclimatized for 1 week before specimen collection.Prior to specimen collection,all groups of mice were fasted for 24 hours,and 0.2 mL of olive oil was administered by gavage to each group of mice at the end of the fasting period,followed by gavage of 0.2 mL of semi-solid nutrient solution 30 minutes before specimen collection.Small intestine propulsion tests were conducted to assess the small intestine motility of each group of mice.Biochemical reagent kits were used to measure the serum levels of triglycerides(TG),total cholesterol(TC),and low-density lipoprotein(LDL) in each group of mice.Transmission electron microscopy was used to observe changes in the number of lipid droplets in intestinal epithelial cells of each group.The calcium cobalt method was used for alkaline phosphatase staining to detect intestinal alkaline phosphatase(IAP) activity in each group of intestines.Hematoxylin-eosin(HE) staining was used to observe the morphology of intestinal epithelia in each group.Immunohistochemical staining was performed to detect the expression of Olfm4,Lgr5,PCNA,Notch1,Hes1 and Villin in the small intestines of each group of mice.Western blot analysis was conducted to determine the expression levels of lipid absorption-related molecular proteins in the small intestines of each

关 键 词：桂附地黄丸 衰老 脂质吸收 小肠 肠吸收细胞 

分 类 号：R285.5[医药卫生—中药学]