补肝散调节R-spondin1/Wnt3a介导的肠道损伤改善D-半乳糖致衰老大鼠消化吸收功能障碍  

Bugansan Regulates R-spondin1/Wnt3a-mediated Intestinal Injury to Ameliorate Digestion and Absorption Dysfunction in Rat Model of Aging Induced by D-galactose

作　　者：王翼轩 霍冉 田津 方芳 刘娜 王杰鹏 方朝义 WANG Yixuan;HUO Ran;TIAN Jin;FANG Fang;LIU Na;WANG Jiepeng;FANG Chaoyi(Hebei University of Chinese Medicine,Shijiazhuang 050200,China;Hebei Key Laboratory of Integrated Chinese and Western Medicine for Lung Diseases Research,Shijiazhuang 050091,China)

机构地区：[1]河北中医药大学,石家庄050200 [2]河北省中西医结合肺病研究重点实验室,石家庄050091

出　　处：《中国实验方剂学杂志》2025年第9期19-27,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：河北省高等学校科学技术研究项目(ZD2022043);河北省中医药管理局科研计划项目(2020136);河北中医学院研究生创新资助项目(XCXZZSS2023010)。

摘　　要：目的:基于R-脊椎蛋白1(R-spondin1)/Wnt3a信号途径探讨衰老大鼠消化吸收功能变化与肝虚状态关系及补肝散作用特点。方法:SPF级SD雄性大鼠48只,随机分为空白组,模型组,补肝散低、中、高剂量组,维生素E组,各8只。除空白组腹腔注射生理盐水外,其余各组均腹腔注射D-半乳糖(400 mg·kg^(-1))制备衰老大鼠模型。造模当日起,补肝散低、中、高(7.03、14.06、28.12 g·kg^(-1))剂量组分别灌胃相应剂量水煎液,维生素E组灌胃维生素E混悬液(27 mg·kg^(-1)),空白组和模型组灌胃等量生理盐水(10 mL·kg^(-1))。观察大鼠皮毛色泽、活动情况、体质量、饮水量及摄食量等一般生物学表征变化,结合血清丙二醛(MDA)含量及超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性以评估大鼠衰老状态;测定抓力及血清总胆汁酸(TBA)含量和α-淀粉酶(AMY)活性,以评测大鼠肝气虚损状态;测定十二指肠组织β-半乳糖苷酶(β-gal)活性以评价肠道衰老情况,检测血清胰高血糖素样肽-1(GLP-1)、血管活性肽(VIP)和D-木糖含量以评估消化吸收功能;苏木素-伊红(HE)染色观察大鼠十二指肠病理形态以评估肠道损伤状态;免疫组化染色观察十二指肠B细胞特异性莫洛尼鼠白血病病毒整合位点1(Bmi1)、富含亮氨酸重复序列的G蛋白偶联受体5(Lgr5)表达特征,实时荧光定量聚合酶链式反应(Real-time PCR)检测十二指肠肿瘤增殖细胞核抗原(Ki67)、Bmi1和Lgr5 mRNA表达,以评估增殖与再生情况;Real-time PCR检测十二指肠组织R-spondin1、Wnt3a、β-连环蛋白(β-catenin)、糖原合成酶激酶-3β(GSK-3β)mRNA表达,蛋白免疫印迹法(Western blot)测定十二指肠组织R-spondin1、Wnt3a、β-catenin、p-GSK-3β蛋白表达,以分析衰老大鼠消化吸收功能障发生机制及补肝散作用特点。结果:与空白组比较,模型组大鼠体质量、饮水量、摄食量、抓力及血清SOD、GSH-Px、AMY活性及GLP-1、VIP�Objective:To elucidate the correlation between alterations in digestion and absorption functions and hepatic deficiency states in aging rats based on the R-spondin1/Wnt3a signaling pathway,and reveal the intervention mechanism of Bugansan.Methods:Forty-eight SPF-grade male SD rats were randomly assigned to six groups:blank control,model,low-,medium-,and high-dose(7.03,14.06,28.12 g·kg^(-1),respectively)Bugansan,and vitamin E(suspension,27 mg·kg^(-1)),with 8 rats in each group.The rat model of aging was established by intraperitoneal injection of D-galactose(400 mg·kg^(-1)),while the blank control group was injected with normal saline.Since the day of modeling,rats in intervention groups received corresponding agents by gavage,and those in blank control and model groups received an equal volume of normal saline(10 mL·kg^(-1)).General biological features such as fur color,activity,body mass,water intake,and food intake were observed.Meanwhile,the content of malondialdehyde(MDA)and the activities of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)in the serum were measured to assess aging.Grip strength and the content of total bile acids(TBA)and the activity ofα-amylase(AMY)in the serum were measured to evaluate hepatic deficiency states.The activity ofβ-galactosidase(β-gal)in the duodenum was measured to evaluate intestinal senescence.The levels of glucagon-like peptide-1(GLP-1),vasoactive intestinal peptide(VIP),and D-xylose in the serum were determined to assess digestion and absorption functions of the small intestine.Hematoxylin-eosin staining was conducted to observe pathological changes of the duodenum to assess the small intestine damage.Immunohistochemical staining was employed to visualize the expression of B-cell-specific Moloney murine leukemia virus integration site 1(Bmi1)and leucinerich repeat-containing G protein-coupled receptor 5(Lgr5)in the duodenal tissue.Moreover,Real-time quantitative polymerase chain reaction(Real-time PCR)was utilized to quantify the mRNA levels of Ki67,Bmi1

关 键 词：衰老 消化吸收功能障碍 R-脊椎蛋白1(R-spondin1)/Wnt3a信号通路 肝气虚损 补肝散 

分 类 号：R242[医药卫生—中医临床基础] R289[医药卫生—中医学] R259