Zibo Dong

Liensinine is mainly from lotus seed extraction and separation of alkaloids, a plant-derived isoquinoline alkaloid, modern pharmacology study shows that it has an anti-inflammatory, antioxidant and antidepressant effect, and so on. This study aims to study liensinine in the role of sepsis heart damage model and clarify the therapeutic mechanism of liensinine. In our study, the mice were randomly divided into control group (C), sepsis group (LPS), and liensinine group (L, M, H). Mice in the liensinine group were intraperitoneally injected with 10, 20, and 40 mg/kg liensinine for 7 days, and LPS was injected at 10mg/kg on the 7th day. The animals were euthanized 6 hours after LPS injection, and the heart tissue and serum were removed for subsequent experiments. Our results suggest that liensinine decreases the LPS-induced rise in LDH and CK activity while improving heart shape. In addition, Liensinine pretreatment results in significant downregulation of mRNA levels of LPS-induced pro-inflammatory factors and protein levels of INOS, COX-2, and P-NF-κB p65. Liensinine also triggers the Keap 1/Nrf2 signaling pathway and increases the activity of antioxidant enzymes in the heart. We also found that liensinine reduces the expression of apoptosis key proteins and TUNEL-tagged apoptotic cells. In addition, Liensinine inhibits protein expression of P62 and enhances protein expression of ATG5 and LC3-II/LC3-I ratio, suggesting activation of autophagy in this process. We first show that liensinine has a beneficial effect on LPS-induced cardiac injury and can protect the myocardial structure, inhibit inflammation, oxidative stress, apoptosis, and promote autophagy.

概要 本研究基于脂多糖 (LPS) 诱导的小鼠脓毒症模型, 探讨莲心碱 (LIE) 对脓毒症中脾损伤的潜在保护作用. 脓毒症常伴有炎症、 氧化应激和细胞凋亡, 并将导致身体器官功能障碍, 对脾脏损伤尤甚. 将 30 只小鼠随机分为 5 组 (n=6): 对照组、 LPS (10 mg/kg)、 LIE (10 mg/kg)+LPS、 LIE (20 mg/kg)+LPS 和 LIE (40 mg/kg)+LPS. 脾脏损伤通过苏木精–伊红染色 (H&E) 确定; 采用定量聚合酶链反应 (qPCR) 检测脾脏组织中肿瘤坏死因子 α (TNF-α)、 白细胞介素 6 (IL-6)、 IL-10、 IL-1β 和一氧化氮合酶 (iNOS) 的转录水平. 同时对包括丙二醛 (MDA)、 过氧化氢酶 (CAT)、 超氧化物歧化酶 (SOD) 和谷胱甘肽过氧化物酶 (GSH-Px) 在内的氧化应激指标进行测定; 采用 TUNEL 法检测细胞凋亡水平. 结果显示, LIE 可减轻脾脏组织病理学损伤并抑制细胞凋亡, 显著降低促炎因子 TNF-α、 IL-6、 IL-1β 和 iNOS 的转录水平, 并增加抗炎因子 IL-10 的表达. 此外, LIE 预处理后可降低 MDA 脂质过氧化指标, 增强 CAT、 SOD和 GSH-Px 的抗氧化活性. 综上所述, LIE 具有潜在的抗氧化和抗炎能力, 可减轻脾脏在脓毒症中的损伤.

Aim The method of D-limonene of Fructus Aurantii Immaturus volatile oil clathrates (clathrates) was established and the solubility of clathrates was determined. Methods: The detection method of clathrates was HPLC, and the solubility of clathrates was determined by equilibrium method. Results Welch UItimate XB-C18 chromatographic column (4.6 mm×250 mm, 5 μm); The detection wavelength: 205 nm; column temperature: 35℃; sample size: 10 μL; mobile phase: methyl cyanide (A)-water (B); current speed: 1 mL/min, gradient elution program: 0~10 min 65%~75% ( A); 10~28 min 75%~78% (A); 28~32 min 78%~65% (A); 32~42 min 65% (A). The solubility of the clathrates in water is 12.75 times that of the volatile oil. Conclusion HPLC method of D-limonene content and inclusion rate is accurate and reliable, it can be used as the method to measuring the rate of clathrates. And this experimental method is stable and reliable, can prepare the clathrates with good solubility and stable properties.