AbstractTamarix tetragyna is a plant grows in Mediterranean area and some Arab countries. It possesses numerous medicinal values. Purpose of our study is to explore biological activity of tamarix tetragyna extracts of both leaves and stem with investigating their phytochemical composition. The investigated extracts’ phyto-constituent composition was determined using gas chromatographic-mass spectrometric method.

Tamarix tetragyna是一种生长在地中海地区和一些阿拉伯国家的植物。它具有许多药用价值。本研究的目的是探索柽柳叶和茎提取物的生物活性，并研究其植物化学成分。使用气相色谱-质谱法测定所研究提取物的植物成分组成。

In addition, in vitro cytoxicity activity versus cancer cell lines such MCF-7, HepG-2, HCT-116, and A-549 was examined by MTT assay method, together with exploring its apoptosis effect by flow cytometry and western blot analysis techniques. Moreover, some phytochemical compounds were identified, and in-silico evaluated against anticancer molecular targets.

此外，通过MTT测定法检测其对MCF-7，HepG-2，HCT-116和A-549等癌细胞系的体外细胞毒性活性，并通过流式细胞术和蛋白质印迹分析技术探讨其凋亡作用。此外，鉴定了一些植物化学化合物，并针对抗癌分子靶标进行了计算机评估。

Plant extracts showed good cytotoxic activity against both A-549 and HCT-116 cancer cell lines. With an IC50 value of 23.90 µg/ml that led to apoptosis and G2/M-phase arrest in A-549 cells, cytotoxicity data demonstrate leaves’ extract effectiveness against these cells. Upon GC-MS analysis, it revealed presence of some bioactive components such as Stigmast-5-en-3-ol and 2-methoxy-4-vinyl phenol, which are known for their cytotoxic activity.

植物提取物对A-549和HCT-116癌细胞系均显示出良好的细胞毒活性。IC50值为23.90µg/ml，导致A-549细胞凋亡和G2/M期阻滞，细胞毒性数据表明叶片提取物对这些细胞的有效性。经GC-MS分析，发现存在一些生物活性成分，如豆甾-5-烯-3-醇和2-甲氧基-4-乙烯基苯酚，它们以其细胞毒活性而闻名。

Our findings suggest that methanolic extracts of Tamarix tetragyna parts may have potential therapeutic uses as anticancer against A-549 cells, which opens up further avenues for investigation into its industrial applications..

我们的研究结果表明，柽柳部分的甲醇提取物可能具有抗A-549细胞的潜在治疗用途，这为研究其工业应用开辟了进一步的途径。。

IntroductionUse of plants as a source of medicine has been inherited and is an important component of the health care system. Numerous medicinal plants have been utilized for ages in traditional medicine to cure a variety of diseases1. A wide variety of chemicals in medicinal plants can be used to treat both chronic and infectious disorders2.

。许多药用植物在传统医学中已被用于治疗多种疾病1。药用植物中的多种化学物质可用于治疗慢性和传染性疾病2。

Several plant extracts have been shown to have a high therapeutic activity. These extracts were made utilizing a variety of organic solvents, including ethanol, methanol, and acetone3.More than 60 species of halophyte plants belonging to the genus Tamarix of the Tamaricaceae family, often known as “Tamarisk” and “salt cedar,” are cultivated all over the world.

几种植物提取物已被证明具有很高的治疗活性。这些提取物是利用多种有机溶剂制成的，包括乙醇，甲醇和丙酮3。属于柽柳科柽柳属的60多种盐生植物，通常被称为“柽柳”和“盐雪松”，在世界各地种植。

These plants have salt-coated needle-like leaves that are produced by salt glands and are covered in salt4. Although tamarisk species are best recognized for flourishing in hot, arid areas, they can also be found there5. Tamarix species are grown in dry regions to aid in stabilizing sand dunes6, but their development in moist climates is prohibited because they act as invasive plants that hinder the development of other species7.

这些植物有盐渍的针状叶子，由盐腺产生，被盐覆盖。虽然柽柳品种在炎热干旱地区最为常见，但它们也可以在那里找到5。柽柳物种生长在干旱地区，以帮助稳定沙丘6，但禁止在潮湿气候下生长，因为它们是阻碍其他物种发育的入侵植物7。

Tamarix tetragyna is a perennial shrub with height of about 2.76 m. Its flowering period is between March and October every year8.The strong antioxidant system found in Tamarix species is a result of particular phenolic chemicals, terpenoids (carotenoids and essential oils), and vitamins. These elements are required for plants to regularly grow, develop, and guard against damage and infection9.

柽柳（Tamarix tetragyna）是一种多年生灌木，高度约为2.76米。它的花期是每年的3月至10月8。柽柳中发现的强抗氧化系统是特定酚类化学物质，萜类（类胡萝卜素和精油）和维生素的结果。这些元素是植物正常生长，发育和防止损害和感染所必需的9。

Along with their many therapeutic advantages, these compounds have the ability to function as antioxidants and antibacterial agents, as well as hepato-protective and chemo-preventive properties, anti-inflammatory, anti-allergic, antithrombotic, cardio-protecti.

这些化合物除了具有许多治疗优势外，还具有抗氧化剂和抗菌剂的功能，以及保肝和化学预防特性，抗炎，抗过敏，抗血栓形成，心脏保护作用。

T. tetragyna methanolic extract of leaves induced G2/M phase cell-cycle arrest in A-549 cellsAs leaves extract’s cytotoxic activity was shown to be highest against the A-549 cancer cell lines, we aimed to characterize bioactivity of this extract by showing its impact on the progression of cell cycle.

T、 叶的四倍体甲醇提取物诱导A-549细胞G2/M期细胞周期停滞。叶提取物对A-549癌细胞系的细胞毒活性最高，我们旨在通过显示其对细胞周期进程的影响来表征该提取物的生物活性。

A-549 cancer cells treated with a methanolic extract of leaves showed an increase in the percentage of cells in the G2/M phase (18.62% compared to 12.81% in untreated cells), as shown in Fig. 4. These findings indicate that percent of A-549 cancer cells that arrested in G2/M phase has been increased in presence of TML which reveals inability for cell division.

用叶的甲醇提取物处理的A-549癌细胞显示G2/M期细胞百分比增加（18.62%，而未处理细胞为12.81%），如图4所示。这些发现表明，在存在TML的情况下，停滞在G2/M期的A-549癌细胞的百分比增加，这表明无法进行细胞分裂。

Hence we can conclude that TML has induced apoptosis in A-549 cells. Thus, cell cycle analysis revealed G2/M-phase arrest and activation of apoptosis in A-549 cells.Fig. 4Cell-cycle histograms showing effect of TML extract on cell-cycle progression in A549 cancer cells after 72 h of treatment at IC50 concentration (B) compared to cell-cycle histogram of blank A-549 cells (A).Full size image.

因此，我们可以得出结论，TML已诱导A-549细胞凋亡。因此，细胞周期分析显示A-549细胞中G2/M期阻滞和凋亡激活。图4细胞周期直方图显示，与空白A-549细胞的细胞周期直方图（A）相比，在IC50浓度（B）处理72小时后，TML提取物对A549癌细胞中细胞周期进程的影响。全尺寸图像。

T. tetragyna methanolic extract of leaves activated caspase-dependent and bax and p53 mediated apoptosis in A-549 cellsAnalysis of the cell cycle revealed that methanolic leaf extract led to apoptosis in A-549 cells. As a result, an annexin V/propidium iodide (PI) apoptosis assay was performed using leaves’ extract on A-549 cell lines to confirm that.As cells in late apoptosis or necrosis (both annexin and PI positive) are represented by Q2 quadrant.

T、 叶的tetragyna甲醇提取物激活了A-549细胞中caspase依赖性和bax和p53介导的细胞凋亡细胞周期分析表明，甲醇叶提取物导致A-549细胞凋亡。结果，在a-549细胞系上使用叶子提取物进行膜联蛋白V/碘化丙啶（PI）凋亡测定以证实这一点。由于晚期凋亡或坏死的细胞（膜联蛋白和PI阳性）由Q2象限表示。

Hence, leaves’ extract induced late apoptosis (8.66% in Q2 compared to 0.27% only in untreated cells). As cells in early apoptosis or cell apoptosis (annexin positive and PI negative) are represented by Q4 quadrant, therefore, leaves’ extract induced early apoptosis (19.53% in Q4 compared to 0.43% only in untreated cells).

因此，叶片提取物诱导晚期细胞凋亡（Q2为8.66%，而未处理细胞仅为0.27%）。由于早期凋亡或细胞凋亡（膜联蛋白阳性和PI阴性）的细胞由Q4象限表示，因此，叶片提取物诱导早期凋亡（Q4为19.53%，而未处理细胞为0.43%）。

In addition, TML extract caused more necrosis that is confirmed by its high percentage (4.32%) compared to that of untreated cells (1.58%) in Q1 quadrant, as shown in both Table 4 and Fig. 5.Fig. 5Apoptosis quadrant plots showing apoptotic effects of TML extract on A549 cancer cells upon treatment at IC50 concentration for 72 h.Full size imageTherefore, we may conclude that the methanolic extract of the leaves caused the death of cancer cells, primarily via inducing apoptosis with a small amount of necrosis.Table 4 Apoptotic assay of TML extract on lung cancer cell lines (A-549).Full size tableProteins called caspases are primarily in charge of apoptosis.

此外，TML提取物引起更多的坏死，与Q1象限中未处理细胞（1.58%）相比，其高百分比（4.32%）证实了这一点，如表4和图5所示。图5凋亡象限图显示了TML提取物在IC50浓度处理72小时后对A549癌细胞的凋亡作用。全尺寸图像因此，我们可以得出结论，叶子的甲醇提取物主要通过诱导细胞凋亡和少量坏死而导致癌细胞死亡。表4 TML提取物对肺癌细胞系（A-549）的凋亡测定。称为半胱天冬酶的全尺寸表蛋白主要负责细胞凋亡。

Caspase-3 is one of caspases that can be considered as a frequently activated death protease. As shown in caspase-cascade system, activation and function of caspases are organized by different types of molecules, such as Bcl-2 family proteins, calpain, apoptosis protein inhibitor and Ca2+48. One of the proteins involved in tumor suppression and subsequen.

半胱天冬酶-3是一种半胱天冬酶，可被认为是一种频繁激活的死亡蛋白酶。如半胱天冬酶级联系统所示，半胱天冬酶的激活和功能由不同类型的分子组织，例如Bcl-2家族蛋白，钙蛋白酶，凋亡蛋白抑制剂和Ca2+48。参与肿瘤抑制和随后的蛋白质之一。

Data availability

数据可用性

All data generated or analysed during this study are included in this published article.

本研究期间生成或分析的所有数据均包含在本文中。

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Download referencesFundingThis work was supported by Scientific Research Deanship at University of Ha’il – Saudi Arabia, through project number (RG-23 001).Author informationAuthors and AffiliationsDepartment of Pharmaceutical Chemistry, College of Pharmacy, University of Hail, Hail, 81442, Saudi ArabiaBader Huwaimel, Kareem M.

下载参考文献资助这项工作得到了沙特阿拉伯哈伊尔大学科学研究院长的支持，项目编号为（RG-23 001）。作者信息作者和附属机构Hail大学药学院药物化学系，Hail，81442，沙特阿拉伯Huwaimel，Kareem M。

Younes & Amr S. AbouziedMedical and Diagnostic Research Centre, University of Ha’il, Hail, 55473, Saudi ArabiaBader HuwaimelDepartment of Biochemistry, Faculty of Pharmacy, University of Sadat City, Cairo, EgyptAkram M. ElkashlanDepartment of pharmaceutics, College of Pharmacy, University of Ha’il, Hail, Saudi ArabiaFawaz N.

Younes＆Amr S.AbouziedMedical and Diagnostic Research Centre，哈伊尔大学，Hail，55473，沙特阿拉伯阿卜杜勒·胡瓦伊梅尔（Huwaimel）Sadat City大学药学院生物化学系，开罗，埃及阿克兰M.ElkashlandDepartment of pharmaceutics，哈伊尔大学药学院，沙特阿拉伯哈伊尔N。

Alheibshy & Ahmed AlobaidaDepartment of Pharmaceutics, Faculty of Pharmacy, Aden University, 6075, Aden, YemenFawaz N. AlheibshyCollege of Pharmacy, University of Ha’il, Hail, 81442, Saudi ArabiaAbdullah Turki, Saleh A. Alquwaiay, Naif Alqahatani & Sulaiman A. AlsuwayaghAuthorsBader HuwaimelView author publicationsYou can also search for this author in.

亚丁大学药学院药剂学系Alheibsy＆Ahmed AlobaidaDepartment of Pharmaceutics，亚丁大学药学院，6075，YemenFawaz N.哈伊勒大学药学院，海尔，81442，沙特阿拉伯阿卜杜拉图尔基，萨利赫A.阿尔库瓦伊，奈夫·阿尔卡哈塔尼和苏莱曼A.阿尔苏瓦亚·加乌托尔斯巴德·胡瓦伊梅尔维尤作者出版物你也可以在中搜索这位作者。

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PubMed Google ScholarContributionsBader Huwaimel: Formal analysis, Investigation Kareem M. Younes: Conceptualization, Writing – original draft. Amr S. Abouzied: review & editing, docking study. Akram M. Elkashlan: Investigation, Methodology, Funding acquisition. Fawaz N. Alheibshy: review & editing.

PubMed Google ScholarContributionsBader Huwaimel：形式分析，调查Kareem M.Younes：概念化，写作-原稿。Amr S.Abouzied：评论与编辑，对接研究。Akram M.Elkashlan：调查，方法，资金获取。Fawaz N.Alheibsy：评论与编辑。

Ahmed Alobaida: Formal analysis, Investigation, Abdullah Turki: Methodology. Saleh A. Alquwaiay: Methodology. Naif Alqahtani: Methodology. Sulaiman A. Alsuwayagh: Methodology.Corresponding authorCorrespondence to.

艾哈迈德·阿洛拜达：形式分析，调查，阿卜杜拉·图尔基：方法论。萨利赫·阿尔库瓦伊：方法论。Naif Alqahtani：方法论。Sulaiman A.Alsuwayagh：方法论。对应作者对应。

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Reprints and permissionsAbout this articleCite this articleHuwaimel, B., Younes, K.M., Abouzied, A.S. et al. Phytochemical composition, in vitro cytotoxicity, and in silico docking properties of Tamarix tetragyna L..

转载和许可本文引用本文Huwaimel，B.，Younes，K.M.，Abouzied，A.S。等人。柽柳的植物化学成分，体外细胞毒性和计算机对接特性。。

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