Citrus aurantium L. extract alleviate depression by inhibiting gut microbiota-mediated inflammation in mice

Gut microbiota plays a crucial role in the pathophysiology of depression.This study aimed to explore the antidepressant effect of mature whole Citrus aurantium fruit extract(FEMC)in the chronic unpredictable mild stress(CUMS)model.The behavioral tests were applied to assess antidepressant effect and 16S rRNA sequencing was used to analyze the changes of gut microbiota.The results showed that the major components of FEMC were naringin and neohesperidin and significantly increased the sucrose preference index of the mice.FEMC also could reduce the feeding latency in an open field test and the rest time in a novelty suppressed feeding test.In addition,FEMC could increase CUMS-induced reduction in the levels of BDNF,PSD95,and SYN in the hippocampus.Moreover,FEMC intervention slightly decreased the ratio of Firmicutes to Bacteroidota.Meanwhile,FEMC reduced the abundance of the Prevotellaceae_Ga6A1_group,[Ruminococcus]_torques_group,which have been reported to be closely related to inflammation.Bioinformatics analysis revealed that mitogen-activated protein kinase(MAPK)signaling pathway and lipopolysaccharide biosynthesis were involved in the anti-inflammatory effect of FEMC in the CUMS animal model.Finally,the ELISA results showed that FEMC could significantly reduce the expression of pro-inflammatory cytokines IL-6 and TNF-αin the serum of depressive mice.Our results suggest FEMC can am eliorate depressive behavior by i nhibiting gut microbiota-mediated inflammation in mice.

Effect of <i>Citrus aurantium</i>L. Essential Oil and Its Interaction with Fluoxetine on Anxiety in Male Mice

Anxiety is a very common mental disorder among neurological diseases. Some herbs have soothing effects and play an important role in reducing anxiety. The purpose of this study is to investigate the effect of Citrus aurantium L. essential oil on anxiety and its interference with serotonergic pathway. Sixty male mice were assigned into control, sham (saline and olive oil), and experimental groups. Intraperitoneal injection of Citrus aurantium L. essential oil was applied at doses of 0.5, 2.5, and 5 percent for 5 days. In another set of experiments, after intraperitoneal injection of Citrus aurantium L. essential oil at doses of 0.5, 2.5, and 5 percent for 5 days, on the 5th day, 30 minutes before applying essential oil, fluoxetine (2 mg/kg) was injected. Then, the anxiety-related behavior was assessed using elevated plus maze test. The results revealed that injection of essential oil of Citrus aurantium L. alone or along with fluoxetine led to increasing the number of entries into the open arms and the time spent in open arms that was significantly different compared with control and sham groups (P?< 0.001). Besides, further effects revealed when fluoxetine added to essential oils, however no more effects obtained when compared to fluoxetine alone. It is concluded that Citrus aurantium L. essential oil can reduce the anxiety in male mice and due to fluoxetin potentiation and maximum response observed, the herb may express its anxiolytic effects in part, via serotonergic system.

A Study of the Effect of <i>Citrus aurantium</i>L. Essential Oil on Anxiety and Its Interaction with GABAergic Pathways in Male Mice

Anxiety is among the most common disorders affecting a great number of people. Different neurotransmitter systems, such as GABAergic system, play an important role in emergence of anxiety-related behaviors. In this study, we have investigated the effect of Citrus aurantium L. essential oil on anxiety and its interaction with GABAergic pathways. Male mice were assigned into control, sham, and experimental groups. Intraperitoneal injection of Citrus aurantium L. essential oil was applied at doses of 0.5, 2.5, and 5 percent for 5 days. Diazepam (0.1 mg/kg) was injected on the fifth day, thirty minutes before applying Citrus aurantium L. essential oil. The anxiety-related behavior of mice was then assessed using elevated plus-maze test. In groups receiving Citrus aurantium L. essential oil at doses of 2.5 and 5 percent, the percent time spent in the open arms increased significantly (P Citrus aurantium L. essential oil resulted in increasing in the number of entries to the open arms and the percent time spent in the open arms (P Citrus aurantium L. essential oil can reduce anxiety-related behaviors in male mice thah may act via GABAergic system.

Effect of <i>Citrus aurantium</i>L. Essential Oil and Haloperidol on Anxiety in Male Mice

Relationship between sociability and the amount of brain’s dopamine is very well known. In this study, we have examined the effect of Citrus aurantium L. essential oil on anxiety and its interaction with dopaminergic pathways. 70 male mice were assigned into experimental, control, and sham groups. Essential oil of Citrus aurantium L. was injected intraperitonealy at doses of 0.5%, 2.5% and 5% for 5 days. Subcutaneous injection of haloperidol was administered on the fifth day, 30 minutes before the injection of the essential oil. The anxiety-related behavior of mice was then assessed by elevated plus-maze test. The result of this study showed that the injection of Citrus aurantium L. essential oil at doses of 2.5% and 5% increased significantly the time spent in the open arms (OAT) (p < 0.001), also there was a significant increase in the number of entries into the open arms (OAE). Injection of different doses of the essential oil along with haloperidol significantly increased OAT (p < 0.001(. The results demonstrate that the essential oil of Citrus aurantium L. along with haloperidol medication reduces anxiety-related behaviors.

Interaction of Flomazenil with Anxiolytic Effects of <i>Citrus aurantium</i>L. Essential Oil on Male Mice

Due to our previous findings about the role of GABAegic neurotransmission in anxiolytic effects of Citrus aurantium L. essential oil, we are now presenting flomazenil interaction with this herb, as an antagonist of benzodiazepines at GABA receptor. The study was performed on 84 male albino mice assigned to 14 groups of six. The animals were injected intraperitoneally with the Citrus aurantium L. essential oil for 5 days. On the fifth day, either normal saline or flomazenil (0.1 mg/kg) was injected to the experimental groups. Thirty minutes after the injection, all the groups were assessed for anxiety-related behavior by elevated plus-maze test. In groups receiving Citrus aurantium L. essential oil at doses of 2.5 and 5 percent, the time spent in the open arms increased significantly (P aurantium L. essential oil regarding the increment in the number of entries to the open arms and the time spent in the open arms (P aurantium L. essential oil induces its anxiolytic effects like benzodiazepines, in the same site at GABA receptor.

Protective Effect of Immaturue Bitter Orange(Citrus aurantium L.)Flavonoids Extracts on PC12 Cell Injury Induced by 6-Hydroxydopamine

To study the neuro protective effect of flavonoids extracts from immature bitter orange(Citrus aurantium L.),the PC12 cells treated with 6-hydroxydopamine(6-OHDA)were used as the Parkinson’s disease(PD)model.To determine the optimal dose of 6-OHDA for constructing a PD model,PC12 cells were incubated with different concentrations of 6-OHDA for 24 h.After 24 h incubation,PC12 cells of drug groups were added 6-OHDA and different concentrations of flavonoids extracts were measured cell viability by CCK8 for selecting effective concentration of flavonoids extracts;the ROS level was determined using flow cytometry;the levels of MDA,CAT,SOD and GSH-Px were assayed by Colorimetric kit for oxidative stress investigation.Compared with the model group,PC12 cell viability was significantly enhanced(P<0.05),the levels of ROS and MDA were reduced significantly(P<0.05),and the activities of SOD,CAT and GSH-Px were significantly enhanced(P<0.05)in drug groups.In conclusion,immature bitter orange flavonoids extracts could protect PC12 cells against 6-OHDA-induced oxidative stress.

枳壳药材水提液HPLC指纹图谱研究

目的建立枳壳水提液中黄酮类成分特征指纹图谱方法。方法采用回流方法制备枳壳供试品溶液,以乙腈-0.1%甲酸水溶液梯度洗脱,检测波长254nm,建立枳壳水提液的HPLC指纹图谱,对在江西省不同时间采收的枳壳药材进行相似度评价分析。结果从15批不同供试品指纹图谱中确定了12个共有峰,通过与对照品比对,指认了其中的3个特征峰,分别为柚皮苷、新橙皮苷、异柚皮苷。以新橙皮苷峰为参照峰,15批样品指纹图谱中12个共有峰的相对保留时间的相对标准偏差(RSD)均<0.805%。12个共有峰的相对峰面积变化较大,RSD值为196.7%。结论该方法具有较好的重现性和稳定性,不同样品中化学成分的含量变化较大,可对枳壳临床用药及采收期提供指导。

枳壳道地产区主流品种遗传多样性的ISSR分析

采用ISSR分子标记技术对江西、湖南、四川3个枳壳道地产区的主流品种进行遗传多样性分析,为筛选优良品种提供参考。15个引物共扩增114个位点,其中多态性位点108个,多态位点百分率为94.73%;观测等位基因数(Na)1.9467,有效等位基因数(Ne)1.5740;Ne i’s基因多样性(He)33.15%;Shannon’s信息指数(I)0.490 7;UPGMA聚类分析可将所有样品分为4类。枳壳具有较高的遗传多样性,其遗传变异主要存在于不同的品种间;聚类结果与传统的外观形态鉴别分类具有一致性;在进行优良品种选育时,应加强不同品种之间的基因交流,以提高种质的质量。

枳壳活性成分与生态因子的相关性

目的:探讨生态环境对枳壳活性成分形成和累积的影响。方法:采取高效液相色谱法分析了江西樟树、新干,重庆綦江、奉节、江津,湖南沅江、汉寿,浙江衢州等地区枳壳中12种活性成分的含量,利用世界气候数据库及中国土壤数据集获得气温、降水量、海拔等8个生态因子数据,采用典型相关分析等方法探讨生态环境因子对枳壳活性成分的影响。结果:通过对枳壳活性成分含量与生态因子进行典型相关分析,发现采摘期辐射与枳壳黄酮类成分橙皮苷、橘皮素和柚皮苷含量呈显著相关,说明在一定范围内,辐射影响枳壳黄酮类成分累积;而生长期降水量与枳壳香豆素类成分水合橘皮内酯含量呈显著相关,即适宜的降雨有利于水合橘皮内酯含量累积。结论:枳壳种植过程中通过对生长期降水量和采收期辐射的控制可调控枳壳活性成分含量。

柑橘类黄酮成分的同步HPLC检测(英文)

目的:建立检测柑橘类黄酮(柚皮苷、橙皮苷、新橙皮苷、橙黄酮、川陈皮素)的中药材质量控制用快速HPLC方法。方法:用仅含甲醇和水的无盐流动相梯度洗脱,Krom asil反相分析柱(100-5C18-250A,4.6 mm×250 mm,5μm),结合二级阵列管紫外扫描做峰识别。结果:对于要检测的5种柑橘类黄酮均得到满意的分辨率和线性范围,其加样回收率:95%-102%;重现性:RSD1.88%-2.93%:分析了6种酸橙类柑橘中药材黄酮含量。结论:建立了多成分柑橘类黄酮快速HPLC检测方法并可用于LC-MS。

枳壳EST-SSR标记的开发

GenBank上已公布的枳壳EST序列为开发新的SSR标记提供了宝贵的数据资源。本研究利用在线SSR鉴定软件SSRIT分析来自枳壳EST数据库的11029条Unigene序列。分析结果共发现327条EST序列含有348个SSR位点,占总数的2.96%。其中,三核苷酸重复的SSR类型最多,共有161个,占检索总数的46.26%。Primer 3.0设计合成58对EST-SSR引物,其中36对能扩增出产物,6对引物产生多态性分离,分别占所设计引物总数的62.07%和10.34%。本文研究成果为今后枳壳遗传多样性分析、遗传图谱构建及比较基因组等研究方面奠定了基础。

枳实提取液对豚鼠心室肌细胞L型钙电流的影响

目的 :探讨枳实提取液 (CAE)对豚鼠心室肌细胞膜L型钙通道电流 (ICa L)的影响。方法 :用酶解法分离单个心室肌细胞 ,采用全细胞膜片钳技术 ,观察枳实提取液对豚鼠心室肌细胞膜ICa L的影响。结果 :①CAE(4×10 3 ,2× 10 2 ,4× 10 2 ,1× 10 1)g/ml增大ICa L,增加率分别为 15 .89%,2 9.0 1%,5 2 .0 3%和 5 9.76 %(P <0 .0 5 )。②CAE 2× 10 1g/ml和 4× 10 1g/ml分别可使峰值ICa L减少 17.33%和 30 .0 9%(P <0 .0 5 )。CAE只改变电流幅度 ,不改变I V曲线形状。结论 :低浓度CAE ,可浓度依赖性的增大豚鼠心室肌细胞L型钙电流 ,有促进钙通道开放的作用。高浓度CAE ,可抑制心室肌细胞L型钙电流 ,有抑制钙通道开放的作用。

枳壳基因组DNA提取方法的比较研究

以枳壳(酸橙)叶片为试验材料,采用改良CTAB法、CTAB-硅珠法、SDS法、高盐低pH法和周世良法5种方法提取枳壳基因组DNA,并通过紫外分光光度计、琼脂糖凝胶电泳及SSR-PCR扩增3种检测方法比较DNA的提取效果,以期找到枳壳DNA的最佳提取方法。结果表明,5种方法中SDS法提取的DNA浓度最高(平均为900ng/μl),纯度最好(OD260/OD280平均值为1.90),且以SDS法提取的DNA为模板进行SSR-PCR扩增检测,目的条带清晰、亮度均一、稳定性和重复性好。因此,SDS法是枳壳DNA最佳提取方法,可用于分子检测试验。

江枳壳优良类型选择研究

目的 筛选地道药材“江枳壳”优良类型。方法 对江枳壳各类型形态特征、生长发育适应性和结果能力、药材主要药用成分含量比较研究 ,并开展优树选择探讨。结果 江枳壳 6种类型有较明显形态差异 ,生长适应性臭橙较强 ,结实能力香橙较强 ,黄酮类和生物碱成分臭橙较高 ,挥发油成分香橙较高 ,江枳壳单株间药用成分有明显变异 ,选择优树可选育优良品系。

枳壳类中药材干燥果实DNA 提取方法的优化实验设计

以学科前沿为引导,设计了“枳壳类中药材干燥果实DNA提取方法的优化”研究型实验。实验内容包括枳壳类干燥果实DNA的提取、DNA浓度及纯度测定、DNA电泳、PCR产物电泳、结果分析讨论。教学实践表明,该实验有助于学生深入理解干燥中药材DNA提取的全过程,有助于激发学生的自主创新热情,提高学生的科研能力和综合素质。

利用信息熵计算指纹图谱相似度识别江枳壳道地性

目的:以枳壳为模型药物,从信息熵的角度计算指纹图谱的相似度以研究中药的道地性。方法:采用高效液相色谱法分析不同来源枳壳药材指纹图谱;利用信息熵计算指纹图谱相似度识别枳壳道地性并与传统方法进行比较。结果:与传统方法相比较,利用信息熵计算指纹图谱相似度的方法能明显提高对"江枳壳"药材道地性的识别效果。结论:该法为枳壳道地性识别提供了科学的参考依据。

酸橙花和甜橙花的UPLC-Q-TOF-MS/MS定性分析及HPLC含量测定

目的 对酸橙花和甜橙花的化学成分进行定性分析与差异性比较,并对不同产地酸橙花和甜橙花中黄酮类成分进行定量分析。方法 采用UPLC-Q-TOF-MS/MS对酸橙花和甜橙花负离子模式质谱信息进行采集,Peakview质谱分析软件分析数据,通过数据库匹配、质谱峰的裂解规律并结合文献对其成分进行定性分析。质谱数据经MarkerView 1.2.1归一化处理后进行主成分分析与正交偏最小二乘法-判别分析,筛选得到差异性成分。运用HPLC法,采用聚类分析,比较27个不同产地酸橙花和甜橙花中芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷黄酮类成分含量的差异。结果 不同产地酸橙花与甜橙花样品的化学组成差异明显,共鉴定出72个化学成分,47个共有成分中共筛选出9个差异性成分,其中5个黄酮类差异性成分的响应信号表现酸橙花明显高于甜橙花。聚类分析可将27批酸橙花与甜橙花样品按不同产地聚成五类。结论 该研究比较了酸橙花与甜橙花的化学成分组成和指标成分的含量,为江西道地植物酸橙和甜橙资源的综合开发利用提供参考依据。

Box-Behnken响应面法优选枳壳饮片炮制工艺研究

目的:优选枳壳饮片的最佳炮制工艺。方法:以黄酮类成分芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷的含量及总评"归一值"(OD值)为评价指标,采用Box-Behnken响应面法对润透温度、润透时间、干燥温度、干燥时间进行考察,建立枳壳饮片炮制工艺流程标准。结果:最佳炮制工艺为润透温度56℃,润透时间20min,干燥温度49℃,干燥时间4h,筛去瓤核及碎屑。结论:优选的炮制工艺切实可行。

容器规格和缓释肥用量对枳壳扦插育苗的影响

为筛选出对枳壳扦插育苗具有良好效应的容器规格和缓释肥用量,以6月龄的江枳壳秋季嫩枝为插穗,对其进行了A1(5 cm×8 cm)、A2(5 cm×12 cm)和A3(5 cm×16 cm)3种容器规格及B1(2.0 kg缓释肥/m3基质)、B2(2.5 kg缓释肥/m^(3)基质)和B3(3.0 kg缓释肥/m^(3)基质)3个用量处理,通过测定90 d后扦插苗的生根率、生根数、生根力指数、单株根长和叶片数比较了其效果优劣。结果表明:等量缓释肥施用下,容器大小对枳壳扦插苗的观测指标存在显著影响,小容器将限制扦插苗的正常生长,以A2(5 cm×12 cm)为最佳。同一容器大小中,扦插苗的观测指标随着施肥量的增加而提高,但提高幅度变小,以B2(2.5 kg缓释肥/m^(3)基质)最经济。因此,在枳壳扦插育苗生产中,宜采用A2(5 cm×12 cm)白色无纺布网袋作容器和每立方米基质施用2.5 kg缓释肥。