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.

Expressing p20 hairpin RNA of Citrus tristeza virus confers Citrus aurantium with tolerance/resistance against stem pitting and seedling yellow CTV strains

The Citrus tristeza virus (CTV) uses 3 silencing suppressor genes, p20, p23 and p25, to resist the attacks from its Citrus hosts. Inactivating these genes is therefore obviously a potential defensive option in addition to the current control strat-egies including aphid management and the use of mild strain cross protection. In this study, we cloned partial DNA frag-ments from the three genes, and used them to construct vectors for expressing hairpin RNAs (hpRNAs). To facilitate the formation of hpRNAs, the constructs were introduced in a loop structure. Fol owing transformation of sour orange (Citrus aurantium) with these constructs, 8 p20 hpRNA (hp20) and 1 p25 hpRNA (hp25) expressing lines were obtained. The 7 hp20 transgenic lines were further characterized. Their reactions to CTV were tested fol owing inoculation with CT14A and/or TR-L514, both of which are severe strains. Results showed that 3 lines (hp20-5, hp20-6 and hp20-8) were completely resistant to TR-L514 under greenhouse conditions for no detectable viral load was found in their leaves by PCR. However, they exhibited only partial suppression of TR-L514 under screen house conditions since the virus was detected in their leaves, though 2 months later compared to non-transgenic controls. Further tests showed that hp20-5 was tolerant also to CT14A under screen house conditions. The growth of hp20-5 was much better than others including the controls that were concurrently chal enged with CT14A. These results showed that expressing p20 hpRNA was sufifcient to confer sour orange with CTV resistance/tolerance.

Protective effects of a polymethoxy flavonoids-rich Citrus aurantium peel extract on liver fibrosis induced by bile duct ligation in mice

Objective: To evaluate the possible protective effect of Citrus aurantium peel extract(CAE) against apoptosis in cholestatic liver fibrosis induced by bile duct ligation in mice. Methods: Male ICR mice were divided to 5 groups: 1) Control group(Sham-operated mice), 2) Cholestatic liver injury group induced by bile duct ligation(BDL), 3) BDL mice treated with silymarin(200 mg/kg) for 4 weeks, 4) BDL mice treated with 50 mg/kg CAE for 4 weeks, 5) BDL mice treated with 200 mg/kg CAE for 4 weeks. Mice were sacrificed and liver fibrosis was evaluated by serum and hepatic tissue biochemistry tests and liver histopathological examination. Effects of CAE on inflammation and apoptosis gene regulation were investigated through real-time PCR. CAE effect on lipid metabolism related signaling was determined by western blot analysis. Results: In BDL mice, administration of CAE for 4 weeks markedly attenuated liver fibrosis based on histopathological alteration. Serum and hepatic tissue biochemistry results revealed that CAE(50 and 200 mg/kg) decreased the levels of alanine transaminase, aspartate transaminase, gamma glutamyl transferase, total bilirubin, nitric oxide, and thiobarbituric acid reactive substances. Real-time PCR and western blot analysis showed that CAE regulated inflammation, apoptosis, and lipid metabolism factors increased by BDL. Interleukin family, tumor necrosis factor α, and related apoptosis factors m RNA levels were increased by BDL treatment. However, these increases were suppressed by CAE administration. In addition, CAE effectively increased phosphorylation of AMPactivated protein kinase, nuclear factor E2-related factor 2, and related cytoprotective proteins. Conclusions: CAE can efficiently regulate BDL-induced liver injury with antioxidant, antiinflammatory, and antiapoptotic activities.

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.

酸橙夏季光合作用特性研究

[目的]研究酸橙夏季光合作用特性。[方法]采用Li-6400便携式光合作用测定系统测定酸橙不同位置叶片净光合速率、酸橙光合日变化、光响应曲线和CO_(2)响应曲线。[结果]酸橙不同方位和不同部位叶片净光合速率存在一定差异;酸橙夏季净光合速率日变化呈双峰型,存在“午睡”现象,其光合峰值分别出现在10:00和14:30;气孔导度的日变化与净光合速率日变化相似。从光响应曲线看,酸橙光饱和点高,光强为0~2000μmol/(m^(2)·s),净光合速率不断上升,最大为13.44μmol/(m^(2)·s),光补偿点为28.33μmol/(m^(2)·s),表观光量子效率为0.036。从CO_(2)响应曲线看,酸橙CO_(2)饱和点在1500μmol/mol,CO_(2)补偿点为60.65μmol/mol,羧化效率为0.079。相关及回归分析结果表明,夏季酸橙净光合速率与气孔导度、空气相对湿度、胞间CO_(2)浓度、蒸腾速率、水汽压亏缺等生理生态因子关系密切。[结论]酸橙是一种喜阳的C_(3)型植物,且对弱光有较强的适应及利用能力,对CO_(2)具有较强的同化能力,同时在夏季高温低湿环境下,酸橙光合作用受各种生理生态因子影响,此期光合有效辐射不是影响净光合速率的关键因子,空气相对湿度与净光合速率呈显著正相关,因此通过环境选择或增加相对湿度的措施,可缓解酸橙夏季“午睡”现象,提高净光合效率。

酸橙新品种“蜀枳一号”多点品种比较试验

“代代”酸橙(Citrus aurantium‘Daidai’)是传统中药材枳壳(干燥未成熟果实)、枳实(干燥幼果)极其重要的来源。“蜀枳一号”新品种是由广安市邻水县收集到的“代代”优良资源选育而来,于2023年通过四川省农作物品种审定委员会认定。在四川省资阳市、泸州市、广安市和重庆市潼南区开展的品种比较试验表明,“蜀枳一号”有良好的丰产性能,单位面积的枳壳、枳实和代代花(干燥花蕾)产量在所有参试品系和对照品系(广安地方种质)中最高,2020年度分别比对照品系平均增产5.78%、5.44%和5.50%,2021年度分别比对照品系平均增产6.12%、5.60%和5.58%;枳壳的柚皮苷、新橙皮苷含量和枳实的辛弗林、浸出物含量均最高,2020年度分别比对照品系高56.92%、60.46%、35.42%和49.68%,2021年度分别比对照品系高55.05%、86.28%、56.41%和27.99%;枳壳和枳实的水分和总灰分与对照品系无显著差异。“蜀枳一号”产量高,品质突出,在川渝两地具有推广价值。

超声辅助酶法降解枳实果胶及其衍生物抗氧化性能

采用超声辅助酶法对枳实果胶进行降解,获得枳实果胶降解衍生物。以抗氧化活性为指标,考察了超声预处理时间、酶种类、酶添加量等因素对枳实果胶降解衍生物抗氧化活性的影响。利用滴定法、凝胶色谱法和红外光谱等技术分析了枳实果胶降解衍生物中半乳糖醛酸含量、分子量分布以及结构特性,以验证果胶降解衍生物抗氧化构效关系。结果表明:当果胶酶作为降解酶,酶用量为6000 U/g,超声预处理时间为20 min时,制备的枳实果胶降解衍生物的抗氧化活性最佳,此条件下其质量浓度为3 mg/mL时,对DPPH自由基的清除率为62.94%,对ABTS自由基的清除率为52.67%,还原能力为0.9627,与未降解的枳实果胶相比,抗氧化活性显著提高。枳实果胶降解衍生物中半乳糖醛酸含量随降解工艺的不同存在明显差异,且半乳糖醛酸含量与抗氧化活性呈正相关。此外,枳实果胶降解衍生物分子量更低,且甲氧基含量也明显降低,但降解过程并没有改变果胶的吡喃糖结构。研究结果为枳实果胶的高值化利用提供理论依据和参考。

常山胡柚果皮精油提取工艺模型筛选优化、化学成分及抗氧化活性研究

研究常山胡柚果皮(Citrus×aurantium Changshan-huyou pericarp,CP)精油化学成分,并对提取条件进行优化。采用水蒸气蒸馏法提取CP精油,利用气相色谱-质谱法进行分析,通过单因素试验考察不同提取条件对提取率的影响,并利用响应面、人工神经网络进行工艺优化。结果表明,CP精油中主要成分为D-柠檬烯;最佳工艺条件:粒径24目、蒸馏水浸泡1 h、液固比34.86 mL/g、提取温度300.00℃、提取时间166.21 min,在此条件下,CP精油提取率为0.82%,响应面与人工神经网络相结合,可有效地利用已有数据对目标寻优。同时CP精油对DPPH自由基有明显的清除作用(IC_(50)值为65.01 mg/mL),表明CP精油具有一定的抗氧化活性。

不同采收期的酸橙叶片形态及药材质量比较

研究不同采收期酸橙(Citrus aurantium L.)药材干燥后枳实和枳壳的性状特点,比较水分、灰分、浸出物含量和主要化学成分的变化规律,确定酸橙药材枳实和枳壳的最佳采收期,并为充分利用酸橙资源、完善质量控制提供科学依据。采用甲苯法测定水分、热浸法测定浸出物含量,按《中华人民共和国药典(2020年版四部)》规定的方法测定灰分,采用高效液相色谱法测定枳实、枳壳中辛弗林和3种黄酮苷的含量。结果表明,酸橙中果、大果性状稳定性较优,与药典规定规格相近。随着采收期推后,酸橙药材柚皮苷含量呈先升高后降低的趋势,新橙皮苷、橙皮苷、辛弗林含量均呈下降趋势。综合考虑劳动力、时间成本、药材性状、药材有效化学成分等,确定中果酸橙的枳实质量为佳,枳实6月中旬采摘为佳,枳壳7月上中旬果皮尚绿时采收为佳。

枳实麸炒过程中化学成分变化及机理研究

枳实是临床常用的理气药,多用于治疗胃肠道疾病,也可用于治疗咳嗽、气喘等疾病。随着研究的深入,其减肥、升压、兴奋神经的功效逐渐被提及,应用范围从中医药扩展到食品养生、运动保健等多个领域,取得了丰硕的成果。枳实中主要含有黄酮、挥发油、生物碱等成分,而这几种成分大多都具有显著的生理活性,是枳实具有治疗作用的物质基础。枳实经过麸炒后,所含的有效成分发生复杂的变化。文章旨在对枳实炮制前后的化学成分研究现状进行综述,并探讨背后的变化机理,为其他炮制研究提供思路。

创新药枳实总黄酮片的元素杂质研究及控制策略

目的以枳实总黄酮片研发、生产全产业链的元素杂质研究为例,论述中药元素杂质控制策略。方法采用电感耦合等离子体质谱(ICP-MS)法测定枳实总黄酮片的7个元素杂质,经风险评估后,采用《中华人民共和国药典》的重金属检查法测定中间体、饮片的重金属。结果ICP-MS法的专属性、定量限、线性、精密度、准确度等均符合验证要求。测得四批成品的各元素杂质残留量均小于限度的30%;四批枳实提取物的重金属结果均小于10×10^(-6) mg/kg;三批枳实饮片的重金属结果均小于30×10^(-6) mg/kg。结论建立了一套适合中药研发、生产全产业链的元素杂质控制策略。

加味枳实消痞汤联合莫沙必利治疗糖尿病胃轻瘫的临床效果

目的探讨加味枳实消痞汤联合莫沙必利治疗糖尿病胃轻瘫的临床效果。方法选取2022年1月—2023年12月本院收治的糖尿病胃轻瘫患者50例为研究对象,采用双盲抽签法将其分为对照组和观察组,各25例。前者单纯实施莫沙必利治疗,后者则在前者的治疗基础上加用加味枳实消痞汤,观察并对比两组患者治疗前后的胃肠激素变化情况以及临床总有效率。结果治疗前,两组患者胃肠激素各项指标相比,无显著差异(P>0.05);治疗后,观察组胃动素、胃泌素显著低于对照组,生长抑素显著高于对照组(P<0.05)。与对照组相比,观察组临床总有效率显著较高(P<0.05)。结论与单纯莫沙必利治疗相比,联合应用加味枳实消痞汤具有辅助调节胃肠功能的作用,可以提高疗效,改善患者的症状和提高生活质量。

常山胡柚果实不同部位中主要功能性成分分析

本研究针对常山胡柚全果油胞层、白皮层、囊衣、汁胞和籽5个不同部位,连续提取其中的多酚类物质和果胶多糖,采用超高效液相色谱-串联质谱法测定不同部位中的30种柑橘多酚类物质,比较果胶多糖的组成及分子量,全面分析常山胡柚果实不同部位中主要功能性成分的组成。结果表明:常山胡柚各部位黄酮类化合物主要以黄烷酮类形式存在,白皮层中新橙皮苷、新北美圣草苷质量分数较高,分别可达13700.72μg/g(按干质量计,下同)和6270.24μg/g;不同部位酚酸类组成中肉桂酸类含量高于苯甲酸类,且以油胞层中肉桂酸类含量最高;淀粉酶辅助乙二胺四乙酸溶液提取到的白皮层果胶多糖得率最高(27.46%),氢氧化钠(NaOH)溶液提取到的油胞层果胶多糖得率最高(11.30%),且2种提取方法得到的油胞层果胶多糖重均分子量均最大,分别为6.888×10^(5)g/mol和2.343×10^(5)g/mol。常山胡柚油胞层、白皮层和囊衣可作为黄酮类化合物和果胶的加工来源,其丰富的新北美圣草苷可作为胡柚加工副产物新的开发方向。

HPLC测定枳实导滞丸中枳实所含6种成分及其药材基原分析

目的建立枳实导滞丸中芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷、水合橙皮内酯、马尔敏的含量测定方法,同时鉴别其中枳实来源。方法采用Waters Symmetry C 18柱(250 mm×4.6 mm,5μm)色谱柱分析,以乙腈(A)-乙酸水(B)为流动相,梯度洗脱,柱温30℃,流速:1.0 mL·min^(-1),检测波长:285 nm、320 nm,进样量10μL。结果6个成分在各自线性范围内均符合要求,相关系数r均≥0.9993;各成分平均回收率范围在98.68%~101.16%,RSD均≤2.5%。12批枳实导滞丸中芸香柚皮苷质量分数在0.37~5.65 mg·g^(-1),柚皮苷在0~12.16 mg·g^(-1),橙皮苷在0~53.90 mg·g^(-1),新橙皮苷在0~18.90 mg·g^(-1),水合橙皮内酯在0~0.23 mg·g^(-1),马尔敏在0~0.13 mg·g^(-1)。12批次枳实导滞丸样品分别呈现与酸橙或甜橙药材一致的黄酮类成分含量情况。香豆素类成分主要存在于酸橙制备的丸剂中且含量偏低。结论该方法简便、准确、可靠,可同时测定枳实导滞丸中6种有效成分的含量并鉴别丸剂中枳实的基原,可为枳实导滞丸质量评价提供依据。

酸橙花总黄酮的分离纯化及其对抗运动疲劳的影响

从酸橙花中提取总黄酮并对其进行纯化,研究酸橙花总黄酮对抗运动疲劳的影响。结果表明:上样液在总黄酮质量浓度1.5 mg/mL、体积70 mL、pH 5.1、流速2.0 mL/min条件下上样至AB-8大孔树脂后,采用110 mL体积分数72.5%乙醇以1.0 mL/min流速洗脱,所得产物的总黄酮含量由22.12%±1.15%提高至66.42%±1.47%,酸橙花总黄酮回收率为71.9%。抗运动疲劳试验表明,酸橙花总黄酮能够延长小鼠的游泳力竭时间,减少乳酸与尿素氮在小鼠血清中的累积,改善肌酸激酶与超氧化物歧化酶的活力。

药食同源代代花药理作用及应用研究进展

代代花是药食同源目录公布的第一批中药材,含有挥发油类、黄酮类、多糖类、生物碱类、香豆素类等化学成分,具有抗氧化、抗炎、抗菌、抗病毒、抗肿瘤、降血脂等药理作用,在食品、医药、日化等行业有一定应用。作为保健产品组分或中药配方组分,其可起调节机体功能或治疗病症的作用,在大健康领域彰显独特的价值。近年来,代代花的关注度进一步提升,针对其药理作用及应用方面的研究逐渐增加。该研究系统梳理国内外相关研究,从代代花的药理作用以及应用现状等方面进行分析和总结,期望推进深入研究和开发代代花的发展进程,为未来研究和综合开发利用代代花提供理论依据。