背景:沙棘(HRL)的干成熟果实,胡桃科,具有健脾和改善脾虚的传统功能。中医(TCM)诊所已被证明HRL有利于糖尿病的治疗。现代药理研究表明,沙棘总黄酮(TFH)是HRL发挥抗炎和抗糖尿病功能的主要物质。然而,化学特征,HRL的活性成分和抗糖尿病的药理机制仍不清楚。
目的:基于糖尿病并发症的AGE-RAGE信号通路,探讨了TFH抗II型糖尿病(T2DM)的关键靶点和代谢产物。从全面的角度阐述了TFH的抗T2DM机制,包括目标预测,代谢物,潜在的代谢途径,等等。
方法:在本研究中,基于超高效液相色谱-四极杆飞行时间质谱(UPLC-Q-TOF-MS)对HRL的化学成分进行了定性测试。通过网络药理学方法预测HRL的抗T2DM靶点和途径。采用高脂高糖饮食联合链脲佐菌素(STZ)诱导T2DM大鼠模型。通过空腹血糖水平评估T2DM模型,体重,血清生化指标,胰岛素水平和胰岛素抵抗的稳态模型评估。通过代谢物与关键靶标之间的相关性筛选关键代谢途径。最后,通过实验验证了关键靶标和代谢物的定量分析。
结果:TFH干预后,高脂高糖饮食联合链脲佐菌素(STZ)诱导的T2DM大鼠空腹血糖水平显著下调,虽然体重,血清液体水平,胰岛素水平和胰岛素抵抗稳态模型评估(HOMA-IR)得到改善.根据ELISA,蛋白质印迹(WB)和逆转录聚合酶链反应(RT-PCR),TFH显著下调甘油二酯(DAG)激活的蛋白激酶C(PRKCA)的表达水平,丝裂原活化蛋白激酶10(MAPK10),STZ诱导大鼠胰腺中人核因子κB亚基p65(NF-κBp65)和肿瘤坏死因子α(TNF-α)。
结论:TFH下调PRKCA的表达,MAPK10和p65TNF-α以及DAG/PRKCA/MAPK10/TNF-α/p65途径中关键代谢物DA的水平,改善脂质代谢紊乱,抑制炎症反应,从而缓解T2DM的症状。
BACKGROUND: Dry mature fruits of Hippophae rhamnoides L. (HRL), Elaeagnaceae, have traditional functions of invigorating spleen and improving spleen insufficiency. Traditional Chinese medicine (TCM) clinics have been proved that HRL is in favor of diabetes treatment. Modern pharmacological studies demonstrated that total flavones of Hippophae rhamnoides (TFH) are the main substance for HRL to develop anti-inflammation and anti-diabetes functions. However, chemical features, active ingredients and anti-diabetes pharmacological mechanism of HRL still remain unclear.
OBJECTIVE: Key targets and metabolites in anti-type-II diabetes mellitus (T2DM) of TFH have been explored based on AGE-RAGE signaling pathway in diabetic complications. The anti-T2DM mechanism of TFH has been elaborated from comprehensive perspectives, including target prediction, metabolites, potential metabolic pathways, and so on.
METHODS: In this study, a qualitative test of chemical composition of HRL was carried out based on ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). The anti-T2DM targets and pathways of HRL were predicted through network pharmacological approach. The T2DM rat model was induced by high-fat and high-glucose diet combined with streptozotocin (STZ). The T2DM model was evaluated through fasting blood glucose level, body weight, serum biochemical indicators, insulin levels and homeostatic model assessment of insulin resistance. The key metabolic pathways were screened through the correlation between metabolites and key targets. Finally, the quantitative analysis of key targets and metabolites was verified through experiments.
RESULTS: After TFH intervention, the fasting blood-glucose level of T2DM rats induced by high-fat and high-glucose diet combined with streptozotocin (STZ) was downregulated significantly, while body weight, serum liquid level, insulin levels and homeostatic model assessment of insulin resistance (HOMA-IR) were improved. According to ELISA, Western blotting (WB) and reverse transcriptase polymerase chain reaction (RT-PCR), TFH significantly downregulates expression levels of diglyceride (DAG)-activated protein kinase C (PRKCA), mitogen activated protein kinase 10 (MAPK10), human nuclear factor κB subunit p65 (NF-κB p65) and tumor necrosis factor-α (TNF-α) in pancreas of STZ-induced rats.
CONCLUSIONS: TFH downregulates expressions of PRKCA, MAPK10 and p65 TNF-α as well as level of the key metabolite DA in the DAG/PRKCA/MAPK10/TNF-α/p65 pathways, improves lipid metabolism disorder, inhibits inflammatory response and thereby relieves symptoms of T2DM.