基于Wnt/β-catenin信号通路探讨黄芪多糖对脾虚湿困大鼠小肠黏膜损伤修复作用机制

杨彬彬; 崔宁; 王世军

doi:10.14148/j.issn.1672-0482.2022.0077

基于Wnt/β-catenin信号通路探讨黄芪多糖对脾虚湿困大鼠小肠黏膜损伤修复作用机制

Research on the Mechanism of Astragalus Polysaccharides on the Repair of Small Intestinal Mucosal Injury in the Rats with Dampness Stagnancy due to Spleen Deficiency Based on Wnt/β-catenin Signaling Pathway

摘要

摘要:
目的探讨黄芪多糖通过Wnt/β-catenin信号通路修复脾虚湿困大鼠小肠黏膜损伤的机制。
方法 60只Wistar大鼠随机分为对照组、模型组、黄芪多糖高剂量组(900 mg·kg^-1·d^-1)、黄芪多糖中剂量组(600 mg·kg^-1·d^-1)、黄芪多糖低剂量组(300 mg·kg^-1·d^-1)、参苓白术散组(2.5 g·kg^-1·d^-1)。除对照组外, 其余组给予高脂低蛋白饲料加力竭游泳, 连续8周, 建立脾虚湿困大鼠模型。造模成功后, 黄芪多糖及参苓白术散灌胃给药, 持续2周。每周称体质量; 测量各组大鼠小肠推进率; ELISA法检测各组大鼠D-乳酸(D-LA)、二胺氧化酶(DAO)水平; qPCR法检测小肠组织Wnt1、β-catenin、CyclinD1、C-MYC mRNA的表达, Western blot法检测小肠组织Wnt1、β-catenin、CyclinD1、C-MYC蛋白的表达。
结果与对照组比较，模型组体质量显著低于对照组(P < 0.01), 血清D-LA、DAO水平显著升高(P < 0.01), 小肠推进率显著降低(P < 0.01), Wnt1、β-catenin、CyclinD1、C-MYC mRNA及蛋白表达显著升高(P < 0.01);与模型组相比较，黄芪多糖高、中剂量组体质量显著高于模型组(P < 0.05)。黄芪多糖高、中、低剂量组D-LA、DAO显著低于模型组(P < 0.05，P < 0.01), 黄芪多糖高、低剂量组小肠推进率显著升高(P < 0.05，P < 0.01), 黄芪多糖高、中、低剂量组Wnt1、β-catenin、C-MYC蛋白水平显著降低(P < 0.01), 黄芪多糖高、中剂量组CyclinD1蛋白水平显著降低(P < 0.05，P < 0.01), 黄芪多糖高、中、低剂量组Wnt1、β-catenin mRNA表达显著降低(P < 0.01)，黄芪多糖高、中剂量组CyclinD1、C-MYC mRNA水平显著降低(P < 0.01)。
结论黄芪多糖可能通过下调脾虚湿困模型大鼠小肠组织Wnt/β-catenin通路蛋白表达, 修复肠道黏膜损伤, 发挥益气健脾利湿功效的作用。

Abstract:
OBJECTIVE To explore the mechanism of Astragalus polysaccharides (APS) on the intestinal damage in rats with dampness stagnancy due to spleen deficieny, based on Wnt/β-catenin signaling pathway.
METHODS Sixty Wistar rats were randomly divided into six groups: control group, model group, APS high dose group, APS medium dose group, APS low dose group, Shenling Baizhu San (positive drug) group. Except for the control group, the other groups were given high-fat and low-protein feed and exhausted swimming for 8 consecutive weeks to establish a rat model of dampness stagnancy due to spleen deficiency. After successful modeling, APS groups and Shenling Baizhu group were administered the corresponding drugs, respectively, by gavage for 2 weeks. The rats were weighed every week. The propulsion rate of the small intestine of each group were measured. The levels of D-LA, DAO in serum were detected by ELISA. The protein and mRNA expressions of Wnt1, β-catenin, CyclinD1, C-MYC were detected by Western blot and qPCR.
RESULTS Compared with the control group, the rat body weight decreased significantly (P < 0.01), the levels of D-LA, DAO in the serum of the model group significantly increased (P < 0.01), the propulsion rate of small intestine decreased significantly (P < 0.01), the expressions of Wnt1, β-catenin, CyclinD1, C-MYC significantly increased (P < 0.01). After administration intervention, the body weight of APS high and medium dose groups was significantly higher than that of model group (P < 0.05), the levels of D-LA and DAO in APS high, medium and low dose groups were significantly lower than those of model group (P < 0.05, P < 0.01). Compared with the model group, the propotory rate of small intestine significantly increased in APS high, low dose groups (P < 0.05, P < 0.01). The protein and mRNA expressions of Wnt1, β-catenin, CyclinD1, C-MYC in APS high, medium dose groups significantly decreased (P < 0.05, P < 0.01).
CONCLUSION APS can repair intestinal mucosal damage by down-regulating the expressions of proteins related to the Wnt/β-catenin pathway in the small intestine tissue, which may be an important mechanism for its effects of invigorating qi, invigorating the spleen and promoting dampness.

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