背景:鱼苗(Lam。)Aschers花(HPF)以其高类黄酮含量而闻名,这有助于他们广泛作为中药缓解炎症。尽管它们具有公认的潜力,关于HPF的总黄酮(TF)及其在治疗慢性前列腺炎(CP)中的治疗应用的信息仍然未知。
目的:我们旨在研究提取优化,成分分析,TF对CP的缓解作用及其潜在机制。
方法:使用具有Box-Behnken设计模型的响应面方法探索了从HPF中优化TF的提取。基于UHPLC-MS方法鉴定了TF中的主要黄酮。TF的功效(25和100mg/kg,p.o.)在前列腺抗原乳剂诱导的自身免疫性CP大鼠模型中通过测量前列腺指数来评估CP,白细胞和卵磷脂的水平,以及组织病理学检查。蛋白质印迹法检测蛋白质表达量。此外,还在体外评估了TF的抗氧化(DPPH和ABTS)和抗炎(环氧合酶2,COX-2抑制作用)作用。
结果:确定TF提取的最佳条件为60%乙醇浓度,30mL/g液固比,提取时间30min,和90°C的提取温度,提取率为65.98±2.14%。通过与参考标准进行比较来表征TF中总共15种主要类黄酮。TF以剂量独立的方式改善了CP在大鼠中的功效,包括降低的前列腺指数和白细胞水平,卵磷脂身体水平升高,改善前列腺的组织病理学损伤,并抑制核因子κB(NF-κB)p65,NF-κBα(IκBα)抑制剂的磷酸化蛋白表达,c-Jun氨基末端激酶(JNK),p38丝裂原活化蛋白激酶(MAPK),细胞外信号调节激酶(Erk),只是另一种激酶1(JAK1),信号转导和转录激活因子3(STAT3),磷酸肌醇3-激酶(PI3K)和蛋白激酶B(Akt)。同时,TF对DPPH的IC50,ABTS自由基,COX-2分别为2.02、1.79和0.0838mg/mL,分别。
结论:我们首先证明了来自HPF的TF是通过抑制NF-κB来减轻CP的有希望的候选者,MAPK,JAK-STAT,和PI3K-Akt信号通路。
BACKGROUND: Hosta plantaginea (Lam.) Aschers flowers (HPF) are well-known for their high flavonoid content, which contribute to their widely as traditional Chinese medicine for alleviating inflammation. Despite their recognized potential, information regarding the total flavonoid (TF) of HPF and its therapeutic application in treating chronic prostatitis (CP) remains unknown.
OBJECTIVE: We aimed to investigate the extraction optimization, constituent analysis, and alleviating effect of TF on CP as well as its potential mechanism.
METHODS: The optimized extraction of TF from HPF was explored using response surface methodology with a Box-Behnken design model. The major flavonoids in TF were identified based on UHPLC-MS approach. Efficacy of TF (25 and 100 mg/kg, p.o.) on CP was evaluated in prostate antigen emulsion-induced autoimmune CP rat model by measuring prostatic index, the levels of leukocytes and lecithin bodies, as well as histopathological examination. The protein expression contents were detected by western blotting. Additionally, the antioxidant (DPPH and ABTS) and anti-inflammatory (cyclooxygenase 2, COX-2 inhibitory) effects of TF were also evaluated in vitro.
RESULTS: The optimized conditions for TF extraction were determined as 60% ethanol concentration, 30 mL/g liquid-to-solid ratio, 30 min extraction time, and 90 °C extraction temperature, and the extraction ratio is 65.98 ± 2.14%. A total of 15 major flavonoids in TF were characterized by comparison with reference standards. TF ameliorated the efficacy of CP in rats in a dose-independent manner, including reduced prostatic index and leukocytes levels, elevated lecithin body levels, ameliorated histopathological damage to prostate, and suppressed phosphorylated protein expressions of nuclear factor kappa-B (NF-κB) p65, inhibitor of NF-κB alpha (IκBα), c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinase (Erk), just another kinase 1 (JAK1), signal transducer and activator of transcription 3 (STAT3), phosphoinositide 3-kinase (PI3K) and protein kinase B (Akt). Simultaneously, the IC50 of TF to DPPH, ABTS radicals, and COX-2 were 2.02, 1.79, and 0.0838 mg/mL, respectively.
CONCLUSIONS: We first demonstrated that TF from HPF represents a promising candidate to alleviate CP through suppression of NF-κB, MAPKs, JAK-STAT, and PI3K-Akt signaling pathways.