舒胃汤对功能性消化不良大鼠Cajal间质细胞线粒体自噬和细胞凋亡的影响  

Effects of Shuwei Decoction on Mitochondrial Autophagy and Cell Apoptosis of Interstitial Cells of Cajal in Rats with Functional Dyspepsia

作　　者：周滔 梁俊尧 朱莹 徐寅[1] ZHOU Tao;LIANG Junyao;ZHU Ying;XU Yin(The First Hospital of Hunan University of Chinese Medicine,Changsha 410007,China;Hunan University of Chinese Medicine,Changsha 410208,China)

机构地区：[1]湖南中医药大学第一附属医院,湖南长沙410007 [2]湖南中医药大学,湖南长沙410208

出　　处：《中国中医药信息杂志》2025年第2期105-112,共8页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金(81904176);湖南省自然科学基金(2023JJ6004);湖南省教育厅科学研究项目(21B0389);湖南省中医药科研课题(B2023079);湖南省科技创新计划(2021SK51413);湖南省教育厅重点课题(19A375);湖南中医药大学中医学国内一流学科建设项目(2018-2020)。

摘　　要：目的观察舒胃汤对功能性消化不良(FD)肝郁脾虚证大鼠Cajal间质细胞(ICC)钙稳态、氧化应激、线粒体自噬和细胞凋亡的影响,探讨其治疗FD的作用机制。方法将50只SD大鼠随机分为空白组、模型组和舒胃汤低、中、高剂量组(3.78、7.56、15.34 g/kg),每组10只。采用改良复合病因法建立FD肝郁脾虚证大鼠模型,并对舒胃汤低、中、高剂量组大鼠灌胃给药14 d。取胃窦,分离并培养原代ICC,采用Fluo-4 AM、Rhod-2 AM染色分别检测ICC细胞质和线粒体钙离子积累,DCFH-DA、MitoSox Red分别检测ICC细胞质和线粒体活性氧(ROS)含量,免疫荧光染色、流式细胞术分别检测ICC线粒体自噬和凋亡情况,Western blot检测ICC线粒体LC3和电压依赖性阴离子通道(VDAC)1蛋白表达。结果与空白组比较,模型组ICC细胞质和线粒体钙离子、ROS积累均显著增多(P<0.01),TOMM20与LAMP2共表达显著升高(P<0.01),细胞凋亡率显著升高(P<0.01),线粒体LC3Ⅱ/Ⅰ、VDAC1蛋白表达显著升高(P<0.01);与模型组比较,舒胃汤中、高剂量组ICC细胞质和线粒体钙离子、ROS积累均显著降低(P<0.05,P<0.01),TOMM20与LAMP2共表达显著降低(P<0.01),细胞凋亡率显著降低(P<0.01),线粒体LC3Ⅱ/Ⅰ、VDAC1蛋白表达显著降低(P<0.01)结论舒胃汤可能通过降低线粒体钙离子负荷、减少ROS积累及线粒体过度自噬,抑制ICC细胞凋亡,恢复ICC功能,达到治疗FD肝郁脾虚证的目的。Objective To investigate the effects of Shuwei Decoction on calcium homeostasis,oxidative stress,mitochondrial autophagy and cell apoptosis of interstitial cells of Cajal(ICC)in functional dyspepsia(FD)rat with liver depression and spleen deficiency syndrome;To discuss its mechanism in the treatment of FD.Methods Totally 50 SD rats were randomly divided into blank group,model group and Shuwei Decoction low-,medium-and highdosage groups(3.78,7.56,15.34 g/kg),with 10 rats in each group.An improved composite etiology method was used to establish a rat model of FD with liver depression and spleen deficiency syndrome,and rats in the Shuwei Decoction low-,medium-and high-dosage groups were orally administered for 14 days.The gastric antrum was taken,the primary ICC was isolated and cultures,and Fluo-4 AM and Rhod-2 AM staining was used to detect the accumulation of calcium ions in the cytoplasm and mitochondria of ICC,DCFH-DA and MitoSox Red were used to detect the content of reactive oxygen species(ROS)in ICC cytoplasm and mitochondria respectively,immunofluorescence staining and flow cytometry were used to detect mitochondrial autophagy and apoptosis in ICC,Western blot was used to detect ICC mitochondrial LC3 and voltage dependent anion channel(VDAC)1 protein expression.Results Compared with the blank group,the model group exhibited significantly increased accumulation of cytoplasmic and mitochondrial Ca2+and ROS in ICC,the co-localization of TOM20 and LAMP2 markedly increased(P<0.01),along with a significant increase in cell apoptosis rate(P<0.01),the expressions of mitochondrial LC3Ⅱ/Ⅰand VDAC1 proteins significantly increased(P<0.01).Compared with the model group,the accumulation of Ca2+and ROS in ICC cytoplasm and mitochondria was significantly decreased in Shuwei Decoction medium-and high-dosage groups(P<0.05,P<0.01),the co-expression of TOMM20 and LAMP2 significantly decreased(P<0.01),the apoptosis rate decreased(P<0.01),and the expressions of mitochondrial LC3Ⅱ/Ⅰand VDAC1 proteins significantly decrea

关 键 词：功能性消化不良 肝郁脾虚证 舒胃汤 线粒体自噬 细胞凋亡 CAJAL间质细胞 

分 类 号：R285.5[医药卫生—中药学]