槲皮素及其衍生物防治肝损伤作用及机制的研究进展

槲皮素是一种多羟基黄酮类化合物,广泛存在于多种药用植物中,具有抗氧化、抗病毒、抗菌、抗炎和抗肿瘤等多种药理作用。分析了近10年国内外有关槲皮素及衍生物防治肝损伤方面的文献,发现其对各种急、慢性肝损伤均具有良好的防治作用,其作用机制主要涉及抗氧化、抗炎、调节脂质代谢、抗凋亡及自噬、抗病毒等。主要对槲皮素及其衍生物防治不同类型肝损伤的作用及机制进行综述,以期为槲皮素保肝作用的深入研究提供较为全面的文献信息支持。     

基于p38 MAPK信号通路分析槲皮素保护骨性关节炎关节软骨的机制

目的:探讨槲皮素通过靶向干预p38丝裂原活化蛋白激酶(p38 MAPK)信号通路保护膝关节骨性关节炎关节软骨的作用机制。方法:笔者通过网络药理学技术,科学的对槲皮素保护骨性关节炎患者关节软骨的靶点因子、信号通路进行预测,并对其进行分析。选择一条与骨性关节炎相关密切的预测通路对其采用体外细胞实验进行验证,通过细胞计数试剂-8(CCK-8)方法筛选出药物最佳干预浓度,采用酶联免疫吸附测定(ELISA)对骨性关节炎及密切相关炎性因子白细胞介素(IL)-1β,肿瘤坏死因子-α(TNF-α)进行检测,采用实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白免疫印迹法(Western blot)检测槲皮素干预后p38 MAPK信号通路下的相关mRNA,蛋白的表达。结果:通过网络药理学预测MAPK信号通路与骨性关节炎患者(OA)存在十分密切的相关性,并选取该通路中最为密切的p38 MAPK信号通路进行研究,通过药物浓度筛查发现在100μmol·L^-1的槲皮素干预后相关炎性因子降低最为明显,并且,在该浓度的槲皮素干预下抑制p38 MAPK信号通路中p38,磷酸化(p)-p38,基质金属蛋白酶-13(MMP-13),解聚蛋白样金属蛋白酶-4(ADAMTS-4)等关键相关因子表达,促进Ⅱ型骨胶原蛋白(CollagenⅡ)的表达。结论:槲皮素在骨性关节炎疾病防治中具有降低软骨炎性因子表达的作用,可通过抑制p38MAPK通路相关因子表达而向好发展,所有结果显示,槲皮素具有降低OA炎性因子表达,保护OA患者关节软骨的作用。     

槲皮素对低钙高镁饮食大鼠血清矿量和骨代谢的改善作用

目的观察槲皮素对低钙高镁饮食大鼠血清矿量和骨代谢水平的改善作用,为骨质疏松临床预防提供参考。方法将60只清洁雄性SD成年黑色大鼠随机等分为空白对照组、模型对照组和槲皮素喂养组,每组20只。空白对照组实验过程中均正常喂养,模型对照组和槲皮素喂养组均适应性喂养2周后,进行低钙高镁(钙5 mg/L,镁60 g/L,蒸馏水溶液作为饮用水)喂养3个月,槲皮素喂养组给予低钙高镁喂养期间同时给予槲皮素(2.64 kg/d)。观察各组大鼠的日常饮食量、饮水量、体重、毛发、毛发色泽、活动等一般情况,分别于低钙高镁喂养前(实验前)和给药喂养3个月时(实验后)两个时刻点进行血清矿量(血钙、血镁、血磷)及骨代谢指标(BALP、CTX-1、BGP、t PINP)测定。结果实验期间,空白对照组、模型对照组和槲皮素喂养组大鼠的日常饮食量、饮水量、体重比较无显著差异(P0.05),但空白对照组、槲皮素喂养组实验大鼠更具活力,毛发更光润。模型对照组大鼠实验后的血钙量、血磷量、BALP均下降,血镁量、BGP、CTX1、t PINP均提升,与实验前比较差异有统计学意义(P0.05)。槲皮素喂养组大鼠实验后的血钙量、血磷量、BALP均显著高于模型对照组大鼠实验后(P0.05),槲皮素喂养组大鼠实验后的血镁量、BGP、CTX1、t PINP均显著低于模型对照组大鼠实验后(P0.05)。结论槲皮素可显著改善低钙高镁饮食大鼠血清矿量和骨代谢水平,可能在低钙高镁交互作用诱导骨质疏松症的预防中起到重要作用。     

基于Nrf2/Keap1/ARE通路研究槲皮素对小鼠年龄相关性黄斑变性的保护作用

目的：探讨槲皮素通过Nrf2/Keap1/ARE通路对小鼠年龄相关性黄斑变性的保护作用及机制研究。

方法：以昆明小鼠为研究对象,分为：对照组、模型组、槲皮素组; 眼底检查各组小鼠眼底是否出现黄白色似玻璃疣物质; OCT检查各组小鼠视网膜厚度; HE染色观察各组小鼠视网膜形态的改变; FFA观察各组小鼠眼底血管完整性; ELISA检测小鼠血清中SOD、GSH-Px、CAT活性和ROS、MDA含量变化; Western blot检测各组小鼠视网膜中Nrf2/Keap1/ARE相关蛋白的表达情况。

结果：槲皮素能使小鼠眼底的黄白色似玻璃膜疣物质减少且视网膜厚度增加(P<0.05),视网膜血管渗漏点明显减少; 与模型组比较,槲皮素组小鼠的a波振幅、b波振幅较模型组均明显上升(P<0.01); 槲皮素能使小鼠视网膜结构比较清晰,部分外核层发生坏死脱落,且使小鼠血清中ROS、MDA含量降低(均P<0.05),SOD、GSH-Px、CAT活性提高(均P<0.05); 与模型组比较,槲皮素组小鼠视网膜细胞质中Nrf2蛋白表达上调(P<0.05),细胞核中Nrf2蛋白表达下调(P<0.05),小鼠视网膜中Keap1、HO-1、NQO-1、GCL蛋白表达下调(均P<0.05)。

结论：槲皮素可能通过Nrf2/Keap1/ARE通路,改善视网膜光损伤后的氧化应激状态,保护视网膜功能,对年龄相关性黄斑变性具有保护作用。     

高效液相色谱法测定复方矮地茶止咳合剂中的槲皮素和山奈酚含量

目的建立高效液相色谱法测定复方矮地茶止咳合剂中槲皮素和山奈酚的含量。方法采用SUPELCO Discovery C18（4．6mm×250nm,5μm）色谱柱,流动相：甲醇-0．4％磷酸溶液（50：50）,流速：1．0mL·min^-1,检测波长：370nm,柱温：40℃,进样量：10μL。结果槲皮素和山奈酚分别在6．484-155．6μg·mL^-1、5．708～137．0μg·mL^-1与峰面积线性关系良好,低、中、高3个剂量组的平均回收率分别为93．6％、97．2％、98．2％与95．2％、94．9％、100．2％。结论所建立的方法准确可靠、灵敏度高、专属性强,可有效控制复方矮地茶止咳合剂的质量。     

HPLC同时测定芜菁子药材中槲皮素和山奈酚含量及其指纹图谱的研究

目的 建立芜菁子药材中槲皮素和山奈酚定量测定方法及芜菁子中黄酮类化合物的指纹图谱,为芜菁子质量控制提供依据。方法 采用HPLC测定芜菁子药材中槲皮素和山奈酚的含量及建立芜菁子指纹图谱,色谱条件为ODS C₁₈ (4.6 mm×250 mm,5 μm),流动相为甲醇(A)-0.4%磷酸水溶液(B),梯度洗脱,流速0.7 mL·min^-1,检测波长254 nm,柱温30 ℃。结果 样品中槲皮素和山奈酚的含量分别为0.013 1~0.035 6 mg·g^-1及0.022 0~0.048 0 mg·g^-1,平均回收率分别为102.8%和99.5%。芜菁子指纹图谱中有11个共有峰,对10批次芜菁子样品的指纹图谱进行相似度比较,相似度较高。结论 该方法重现性好,可行性强,可用于芜菁子的质量评价。     

Antinociceptive activity of Syzygium cumini leaves ethanol extract on orofacial nociception protocols in rodents

Context: Syzygium cumini (L.) Skeels (Myrtaceae) is a tree with dark purple fruits, popularly known as “jambolão” or “jambolan”. In folk medicine, this plant is used for the treatment of diabetes and inflammatory conditions.

Objective: We investigated the antinociceptive effect of ethanol extract (EE) from S. cumini leaves on orofacial nociception.

Material and methods: The antinociceptive effects of the EE obtained from the leaves of S. cumini were evaluated in mice using formalin- and glutamate-induced orofacial nociception.

Results: ESI-MS/MS analyses demonstrated that major constituents in the analyzed samples coincided with the mass of the phenolic acids and flavonoids. In pharmacological approach, pre-treatment with EE (100, 200, or 400?mg/kg, p.o.) significantly reduced (p?p?G-nitro-l-arginine (l-NOARG). The extract, all doses, also caused a marked inhibition (p?p?Conclusions: We can suggest that the antinociceptive effect of the EE is mediated by peripheral mechanisms, possibly involving K_ATP channels and the nitric oxide pathways. These effects appear to be related to the presence of flavonoids compounds, such as quercetin.     

Effects of quercetin and fish n‐3 fatty acids on testicular injury induced by ethanol in rats

The aim of this study was to investigate the effects of quercetin and fish n‐3 fatty acids on the changes in testis induced by ethanol. Forty‐five rats divided into five groups, control, ethanol, ethanol+quercetin, ethanol+fish n‐3 fatty acids and ethanol+quercetin+fish n‐3 fatty acids. At the end of 8 weeks, all the rats were sacrificed. Degenerative changes in histopathological analyses, the decreased body weight gain and seminiferous tubule diameters in ethanol group have been observed. TUNEL assay also showed an increase in apoptotic cell number. The activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH‐Px), xanthine oxidase (XO) and testosterone levels were decreased as well as the levels of malondialdehyde (MDA) and nitric oxide (NO) were increased in ethanol group. Histopathological changes caused by ethanol have been improved by quercetin and fish n‐3 fatty acids. It was also found that protection was provided by increasing SOD, CAT and GSH‐Px activities in groups administered quercetin, fish n‐3 fatty acids and quercetin+fish n‐3 fatty acids, and by decreasing the levels of MDA and NO in groups administered both quercetin and fish n‐3 fatty acids together. These results suggest that quercetin and fish n‐3 fatty acids are beneficial agents to reduce testicular injury induced by ethanol except for testosterone levels.     

Bioavailability of plant pigment phytochemicals in Angelica keiskei in older adults: A pilot absorption kinetic study

BACKGROUND/OBJECTIVES

Angelica keiskei is a green leafy vegetable rich in plant pigment phytochemicals such as flavonoids and carotenoids. This study examined bioavailability of flavonoids and carotenoids in Angelica keiskei and the alteration of the antioxidant performance in vivo.

SUBJECTS AND MATERIALS

Absorption kinetics of phytochemicals in Angelica keiskei were determined in healthy older adults (> 60 y, n = 5) and subjects with metabolic syndrome (n = 5). Subjects consumed 5 g dry Angelica keiskei powder encapsulated in gelatin capsules with a low flavonoid and carotenoid liquid meal. Plasma samples were collected at baseline, 0.5, 1, 2, 3, 4, 5, 6, 7, and 8 h. Samples were analyzed for flavonoids and carotenoids using HPLC systems with electrochemical and UV detection, respectively, and for total antioxidant performance by fluorometry.

RESULTS

After ingestion of Angelica keiskei increases in plasma quercetin concentrations were observed at 1-3 and 6-8 hr in the healthy group and at all time points in the metabolic syndrome group compared to baseline (P < 0.05). Plasma lutein concentrations were significantly elevated in both the healthy and metabolic syndrome groups at 8 hr (P < 0.05). Significant increases in total antioxidant performance were also observed in both the healthy and the metabolic syndrome groups compared to baseline (P < 0.05).

CONCLUSIONS

Findings of this study clearly demonstrate the bioavailability of phytonutrients of Angelica keiskei and their ability to increase antioxidant status in humans.     

黄酮类成分黄芩素、槲皮素、丹参素钠对吸烟致细胞毒性和DNA损伤的保护作用

目的评价吸烟致细胞毒性和DNA损伤以及黄酮类成分黄芩素、槲皮素、丹参素钠的保护作用。方法以自动吸烟机按照FTC协议吸烟产生的主流烟雾在线染毒B-16细胞和人颊黏膜细胞两种真核细胞,通过MTT比色法和单细胞凝胶电泳法检测吸烟所致的细胞毒性和DNA损伤,并考察黄芩素、槲皮素、丹参素钠的保护作用。结果吸烟可致体外培养的B-16细胞活力明显下降,两种细胞的DNA明显损伤。随着烟气作用时间的延长,表征细胞内DNA损伤程度的彗星尾矩、Olive尾矩都有增加;1 mmol/L槲皮素、黄芩素和丹参素均可明显缓解吸烟引起的细胞毒性和DNA损伤,对B-16细胞的活力提升50%左右,对人颊黏膜细胞的DNA保护效果超过60%。结论吸烟可致细胞毒性和细胞DNA损伤,但是黄酮类成分黄芩素、槲皮素、丹参素钠均对细胞和DNA具有保护作用。