川陈皮素通过激活SIRT1/FOXO3a信号来减轻肾缺血再灌注损伤

目的：本研究旨在探究川陈皮素（NOB）保护小鼠肾缺血再灌注损伤（RIRI）的可能分子机制。方法：将Balb/c小鼠分为5组（n=6）：假手术组、模型组、NOB组（50 mg/kg）、组蛋白去乙酰化酶沉默信息调节因子1（SIRT1）抑制剂EX527组（5 mg/kg）、NOB+EX527组（50 mg/kg的NOB+5 mg/kg EX527）。在建模前24 h对小鼠进行药物处理。通过阻断小鼠左肾动静脉血流建立RIRI模型。建模24 h后检测肾组织中氧化应激标志物含量。通过苏木精-伊红（HE）染色和天狼星红染色评价肾组织病变和纤维化。通过TUNEL染色检测肾细胞凋亡。通过蛋白质免疫印迹法检测肾组织中SIRT1、叉头框蛋白O3a（FOXO3a）、细胞凋亡和自噬相关蛋白表达。通过实时荧光定量PCR检测肾组织中SIRT1和FOXO3a mRNA的水平。结果：与模型组比较,NOB组小鼠肾脏病变程度减轻,肾脏纤维化面积降低（均P<0.05）。与模型组比较,NOB组肾组织抗氧化作用升高（P<0.05）。与模型组比较,NOB组肾组织中细胞凋亡减少（P<0.05）。与模型组比较,NOB组肾组织中SIRT1和FOXO3a的mRNA和蛋白相对表达量升高,微管相关蛋白轻链(LC)3Ⅱ/LC3Ⅰ蛋白相对表达量升高,而p62降低（均P<0.05）。此外,EX527逆转了NOB对肾脏的保护作用（P<0.05）。结论：NOB通过激活SIRT-1/FOXO3a信号通路介导的自噬来减轻RIRI。     

Nobiletin and its derivatives overcome multidrug resistance (MDR) in cancer: total synthesis and discovery of potent MDR reversal agents

Our recent studies demonstrated that the natural product nobiletin (NOB) served as a promising multidrug resistance (MDR) reversal agent and improved the effectiveness of cancer chemotherapy in vitro. However, low aqueous solubility and difficulty in total synthesis limited its application as a therapeutic agent. To tackle these challenges, NOB was synthesized in a high yield by a concise route of six steps and fourteen derivatives were synthesized with remarkable solubility and efficacy. All the compounds showed improved sensitivity to paclitaxel (PTX) in P-glycoprotein (P-gp) overexpressing MDR cancer cells. Among them, compound 29d exhibited water solubility 280-fold higher than NOB. A drug-resistance A549/T xenograft model showed that 29d, at a dose of 50 mg/kg co-administered with PTX (15 mg/kg), inhibited tumor growth more effective than NOB and remarkably increased PTX concentration in the tumors via P-gp inhibition. Moreover, Western blot experiments revealed that 29d inhibited expression of NRF2, phosphorylated ERK and AKT in MDR cancer cells, thus implying 29d of multiple mechanisms to reverse MDR in lung cancer.     

川陈皮素抗肿瘤机制研究进展

作为我国传统中药之一的陈皮，在许多的肿瘤治疗中具有重要的辅助作用。桔皮多甲氧基黄酮类化合物的一种成分川陈皮素，具有抗癌、抗血栓、抗癫痫、抗血凝、抗真菌、抗过敏、抗炎、抗胆碱酯酶等作用。川陈皮素抗肿瘤研究的人越来越多，但其机制仍在研究中，据报道，川陈皮素抗肿瘤机制可能是：首先，抑制肿瘤细胞的生长和增殖；第二，诱导肿瘤细胞凋亡；第三，抑制肿瘤细胞的迁移和侵袭；四是调节癌细胞周期和蛋白表达以及与化疗药物的联合增强抗肿瘤作用有关。本文通过对国内外近10几年来发表的川陈皮素文献进行分析，发现其对多种肿瘤细胞都有抑制作用，将来可能发展为全能型抗肿瘤药物，为川陈皮素在临床抗肿瘤应用中提供参考依据，现就川陈皮素抗肿瘤的相关研究进展进行以下综述。     

Nobiletin对慢性低灌注损伤后小鼠脑海马神经血管单元的影响

目的观察川陈皮素(NOB)对小鼠慢性脑低灌注(CCH)模型海马区神经血管单元各主要组分的保护作用。方法昆明小鼠随机分为对照组、模型组和NOB高、低剂量处理组各10只,模型组和NOB处理组进行双侧颈总动脉结扎,对照组不结扎。术后2 w进行旷场实验及Y迷宫检测学习记忆功能,Western印迹法测定小鼠海马区神经元生长相关蛋白(GAP)-43、突触素(SYN)、水通道蛋白(AQP)4表达,检测海马神经细胞可塑性及血脑屏障功能。结果与模型组相比,NOB处理组自由活动无显著差异,其中高剂量组探索能力显著增强,逃避潜伏期显著减短(P<0.05);NOB处理组均显著增加海马区GAP-43、SYN、AQP4及Claudin-5表达(P<0.05)。结论NOB可通过对海马的神经血管单元各组分的保护作用,改善CCH所致小鼠学习记忆功能受损。     

川陈皮素对肺癌的增殖抑制作用及其机制

目的 观察川陈皮素对肺癌的体内外抑瘤作用并初步探讨其机制.方法 应用MTT法检测不同浓度川陈皮素作用对人肺腺癌细胞A549增殖的抑制作用.Western blot检测川陈皮素处理后A549细胞Bcl-2和Bax蛋白的表达情况.彗星电泳检测川陈皮素作用后A549细胞DNA损伤情况.通过C57BL/6小鼠Lewis肺癌模型进行体内抗瘤实验,TUNEL法观察药物作用后的肿瘤细胞凋亡情况,并用免疫组化法检测瘤组织中Bax、Bcl-2和Caspase-9蛋白表达情况.结果 体外实验显示,川陈皮素作用对A549细胞有明显的增殖抑制作用,并且抑制效应随浓度增加而增强;川陈皮素作用A549细胞24 h后,随着作用剂量的加大,Bcl-2蛋白表达下调,而Bax蛋白则表达上调;对细胞DNA具有损伤作用,当达到一定作用剂量和作用时间时可以使A549细胞发生凋亡.体内抑瘤实验表明,川陈皮素高(300 mg/kg)、中(200 mg/kg)、低(100 mg/kg)浓度组对小鼠Lewis肺癌的抑瘤率分别为43.70%、27.59%和20.14%,与对照组相比,差异具有统计学意义(P<0.01);川陈皮素能使Lewis肺癌细胞发生凋亡;使Lewis肺癌组织内Bax和Caspase-9表达上调,Bcl-2表达下调.结论 川陈皮素在体内外均有明显的抑制肿瘤细胞增殖的作用,其作用机制可能与改变Bcl-2/Bax的比值,启动Caspase级联反应相关.     

Nobiletin improves hyperglycemia and insulin resistance in obese diabetic ob/ob mice

Nobiletin is a polymethoxylated flavone found in certain citrus fruits that exhibits various pharmacological effects including anti-inflammatory, antitumor and neuroprotective properties. The present study investigated the effects of nobiletin on insulin sensitivity in obese diabetic ob/ob mice, and the possible mechanisms involved. The ob/ob mice were treated with nobiletin (200 mg/kg) for 5 weeks. Nobiletin significantly improved the plasma glucose levels, homeostasis model assessment index, glucose tolerance in an oral glucose tolerance test and plasma adiponectin levels. In white adipose tissue (WAT), nobiletin significantly decreased the mRNA expression levels of inflammatory adipokines such as interleukin (IL)-6 and monocyte chemoattractant protein (MCP)-1 and increased the mRNA expression levels of adiponectin, peroxisome proliferator-activated receptor (PPAR)-γ and its target genes. At the same time, nobiletin increased the glucose transporter (Glut) 4 expression levels in the whole plasma membrane, and Glut1 and phospho-Akt expression in the whole cell lysates in WAT and muscle. Nobiletin also increased Glut4 protein expression level in the whole cell lysates of the muscle. Taken together, the present results suggest that nobiletin improved the hyperglycemia and insulin resistance in obese diabetic ob/ob mice by regulating expression of Glut1 and Glut4 in WAT and muscle, and expression of adipokines in WAT.     

Efficacy of nobiletin in improving hypercholesterolemia and nonalcoholic fatty liver disease in high-cholesterol diet-fed mice

BACKGROUND/OBJECTIVESNobiletin (NOB), a citrus flavonoid, is reported to have beneficial effects on cardiovascular and metabolic health. However, there is limited research investigating the effect of long-term supplementation with low-dose NOB on high-cholesterol diet (HCD)-induced hypercholesterolemia and non-obese nonalcoholic fatty liver disease (NAFLD). Therefore, we investigated the influence of NOB on hypercholesterolemia and NAFLD in HCD-fed mice.SUBJECTS/METHODSC57BL/6J mice were fed a normal diet (ND) or HCD (35 kcal% fat, 1.25% cholesterol, 0.5% cholic acid) with or without NOB (0.02%) for 20 weeks.RESULTSHCD feeding markedly reduced the final body weight compared to ND feeding, with no apparent energy intake differences. NOB supplementation suppressed HCD-induced weight loss without altering energy intake. Moreover, NOB significantly decreased the total cholesterol (TC) levels and the low-density lipoprotein (LDL)/very-LDL-cholesterol to TC ratio, and increased the high-density lipoprotein-cholesterol/TC ratio in plasma, compared to those for HCD feeding alone. The plasma levels of inflammatory and atherosclerosis markers (C-reactive protein, oxidized LDL, interleukin [IL]-1β, IL-6, and plasminogen activator inhibitor-1) were significantly lower, whereas those of anti-atherogenic adiponectin and paraoxonase were higher in the NOB-supplemented group than in the HCD control group. Furthermore, NOB significantly decreased liver weight, hepatic cholesterol and triglyceride contents, and lipid droplet accumulation by inhibiting messenger RNA expression of hepatic genes and activity levels of cholesterol synthesis-, esterification-, and fatty acid synthesis-associated enzymes, concomitantly enhancing fatty acid oxidation-related gene expression and enzyme activities. Dietary NOB supplementation may protect against hypercholesterolemia and NAFLD via regulation of hepatic lipid metabolism in HCD-fed mice; these effects are associated with the amelioration of inflammation and reductions in the levels of atherosclerosis-associated cardiovascular markers.CONCLUSIONSThe present study suggests that NOB may serve as a potential therapeutic agent for the treatment of HCD-induced hypercholesterolemia and NAFLD.     

HPLC法同时测定定喘止咳糖浆中5种成分的含量

目的建立高效液相色谱法同时测定定喘止咳糖浆中苦杏仁苷、和厚朴酚、厚朴酚、橙皮苷和川陈皮素含量的方法。方法采用依利特C_(18)柱,流动相为乙腈(A)-0.2%冰醋酸溶液(B),梯度洗脱;流速:0.8 mL/min;柱温:30℃;检测波长:λ_1=210 nm(苦杏仁苷),λ_2=254 nm(和厚朴酚、厚朴酚),λ_3=283 nm(橙皮苷、川陈皮素)。结果苦杏仁苷、和厚朴酚、厚朴酚、橙皮苷、川陈皮素的线性范围分别为12.10~242.00μg/mL(r=0.999 8)、7.65~153.00μg/mL(r=0.999 3)、6.37~127.40μg/mL(r=0.999 7)、4.29~85.80μg/mL(r=0.999 9)、3.90~78.00μg/mL(r=0.999 2);5种成分的平均加样回收率(n=6)分别为98.62%、96.94%、99.17%、96.85%、97.84%,RSD分别为0.88%、1.12%、1.10%、1.04%、1.55%。结论定喘止咳糖浆中5种成分的含量测定方法准确、可靠。