解毒化瘀颗粒含药肠道内容物调控PI3K/AKT信号通路促进大鼠肝上皮样干细胞增殖分化OA
Hepatic Epithelial-Like Stem Cells Proliferation and Differentiation Promoted by Intestinal Contents Containing Jieduhuayu Granules via Intervention of PI3K/AKT Signaling Pathway
目的:基于磷脂酰肌醇 3-激酶(PI3K)/蛋白激酶 B(AKT)信号通路研究解毒化瘀颗粒调节肠道微生态对肝干细胞再生的影响及机制.方法:构建急性肝衰竭大鼠模型,设置正常对照组、模型对照组、解毒化瘀颗粒57.6 g/kg 组,干预48 h 后收集各大鼠肠道内容物悬液并制备肠道内容物总蛋白(IBRP),用于后续干预肝干细胞.体外培养WB-F344 细胞,分为正常对照组、模型对照组、解毒化瘀颗粒0.1、0.2、0.4 mg/mL 含药IBRP 组.采用 CCK-8 法检测不同干预时间下的细胞存活率;倒置生物显微镜下观察肝干细胞形态结构;免疫荧光染色检测白蛋白(ALB);细胞角蛋白 19(CK19)蛋白荧光表达水平;RT-qPCR 法检测 Alb、Ck19 的 mRNA 表达;Western blot 法检测 PI3K、磷酸化 PI3K(p-PI3K)、AKT、磷酸化 AKT(p-AKT)蛋白表达.结果:与正常对照组相比,模型对照组肝干细胞形态明显皱缩,排列紊乱,细胞存活率显著降低(P<0.01),ALB、CK19 蛋白荧光表达水平降低,Alb、Ck19 mRNA 表达明显下调(P<0.05 或 P<0.01),p-PI3K/PI3K 和 p-AKT/AKT 比值显著降低(P<0.01);与模型对照组相比,解毒化瘀颗粒 0.1、0.2、0.4 mg/mL 含药 IBRP 组的肝干细胞形态更加规整,细胞存活率显著升高(P<0.01),ALB、CK19 蛋白荧光表达水平升高,Alb mRNA 表达显著上调(P<0.01),p-PI3K/PI3K 和 p-AKT/AKT比值显著升高(P<0.01).结论:经解毒化瘀颗粒干预后的肠道微生态可以明显促进肝干细胞的增殖分化,并显示出定向转化为成熟肝细胞的趋势,其机制与激活 PI3K/AKT 信号通路有关.
Objective:This paper aims to study the effect and mechanism of Jieduhuayu Granules(解毒化瘀颗粒)regulating gut microbiota to promote hepatic stem cell proliferation via the phosphoinositide 3-kinase(PI3K)/protein ki-nase B(AKT)signaling pathway.Methods:Rat models of acute liver failure were established and divided into a normal control group,a model group,and a Jieduhuayu Granule group.After 48 hours of intervention,rats'intestinal content suspensions from each group were collected to prepare intestinal bacteria-rich protein(IBRP)for subsequent intervention of hepatic stem cells.WB-F344 cells were cultured in vitro and divided into a normal control group,a model group,and Jieduhuayu Granule(0.1 mg/mL,0.2 mg/mL,and 0.4 mg/mL)-containing IBRP groups.Cell viabilities at different intervention times were detected by the CCK-8 assay.The morphological structure of hepatic stem cells was observed un-der an inverted biological microscope.The fluorescence expression levels of albumin(ALB)and cytokeratin 19(CK19)were detected by immunofluorescence staining.The mRNA expressions of Alb and Ck19 were determined by real-time quantitative polymerase chain reaction(RT-qPCR),while the protein expressions of PI3K,phosphorylated-PI3K(p-PI3K),AKT,and p-AKT were detected by Western blot.Results:Compared with those in the normal control group,cells in the model control group showed significant shrinkage and disordered arrangement,with a marked decrease in cell via-bility(P<0.01).The fluorescence expression levels of ALB and CK19 were decreased,and the mRNA expressions of Alb and Ck19 were downregulated(P<0.05 or P<0.01).The ratios of PI3K/PI3K and p-AKT/AKT were significantly reduced(P<0.01).Compared with that of the model control group,the morphology of hepatic stem cells was more reg-ular in the Jieduhuayu Granule(0.1 mg/mL,0.2 mg/mL,and 0.4 mg/mL)-containing IBRP groups,and the cell via-bility was significantly increased(P<0.01).The fluorescence expression levels of ALB and CK19 were increased,and the mRNA expression of Alb was upregulated(P<0.01).The ratios of p-PI3K/PI3K and p-AKT/AKT were significant-ly increased(P<0.01).Conclusion:After intervention with Jieduhuayu Granules,the gut microbiota can obviously pro-mote hepatic stem cell proliferation and differentiation.It shows the trend towards transformation into mature hepatocytes,and its mechanism may be related to activating the PI3K/AKT signaling pathway.
陈研焰;蓝艳梅;王明刚;张荣臻;毛德文
湖南中医药大学,长沙 410208||湖南中医药高等专科学校,株洲 412012广西中医药大学第一附属医院,南宁 530023广西中医药大学第一附属医院,南宁 530023广西中医药大学第一附属医院,南宁 530023广西中医药大学第一附属医院,南宁 530023
解毒化瘀颗粒急性肝衰竭肠道微生态磷脂酰肌醇3-激酶/蛋白激酶 B肝干细胞
Jieduhuayu GranulesAcute liver failureGut microbiotaPhosphoinositide 3-kinase/protein kinase B(PI3K/AKT)Hepatic stem cell
《中药药理与临床》 2026 (7)
68-74,7
广西重点研发计划项目(编号:桂科AB25069020)广西自然科学基金(编号:2025GXNSFDA069037、2023GXNSFAA026176)国家自然科学基金(编号:82274434、82360912、82460901)国家中医药传承创新中心建设项目(编号:2022366).
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