AbstractAcrylamide (ACR) with its extensive industrial applications is a classified occupational hazard toxin and carcinogenic compound. Its formation in fried potatoes, red meat and coffee during high-temperature cooking is a cause for consideration. The fabrication of chitosan-coated probiotic nanoparticles (CSP NPs) aims to enhance the bioavailability of probiotics in the gut, thereby improving their efficacy against ACR-induced toxicity in Drosophila melanogaster.

摘要丙烯酰胺（ACR）具有广泛的工业应用，是一种分类为职业危害毒素和致癌化合物。在高温烹饪过程中，它在炸土豆、红肉和咖啡中的形成是一个值得考虑的原因。壳聚糖包被的益生菌纳米颗粒（CSP-NPs）的制备旨在提高益生菌在肠道中的生物利用度，从而提高其对果蝇ACR诱导的毒性的功效。

Nanoencapsulation, a vital domain of the medical nanotechnology field plays a key role in targeted drug delivery, bioavailability, multi-drug load delivery systems and synergistic treatment options. Our study exploited the nanoencapsulation technology to coat Lactobacillus fermentum (probiotic) with chitosan (prebiotic), both with substantial immunomodulatory effects, to ensure the stability and sustained release of microbial load and its secondary metabolites in the gut.

纳米封装是医学纳米技术领域的一个重要领域，在靶向药物递送，生物利用度，多药物负载递送系统和协同治疗选择中起着关键作用。我们的研究利用纳米封装技术用壳聚糖（益生元）包被发酵乳杆菌（益生菌），两者都具有显着的免疫调节作用，以确保肠道中微生物负荷及其次级代谢产物的稳定性和持续释放。

The combination of pre-and probiotic components, called synbiotic formulations establishes the correlation between the gut microbiota and the overall well-being of an organism. Our study aimed to develop a potent synbiotic to alleviate the impacts of heat-processed dietary toxins that significantly influence behaviour, development, and survival.

pre和益生菌成分（称为合生元制剂）的组合建立了肠道微生物群与生物体整体健康之间的相关性。我们的研究旨在开发一种有效的合生元，以减轻热处理饮食毒素的影响，这些毒素会显着影响行为，发育和生存。

Our synbiotic co-treatment with ACR in fruit flies normalised neuro-behavioural, survival, redox status, and restored ovarian mitochondrial activity, contrasting with several physiological deficits observed in the ACR-treated model..

我们与ACR在果蝇中的合生元共同治疗使神经行为，存活，氧化还原状态正常化，并恢复了卵巢线粒体活性，与ACR治疗模型中观察到的几种生理缺陷形成对比。。

IntroductionGut health has gained considerable attention recently and has become an essential part of most fitness regimes. The gut microbiota, consisting of trillions of microorganisms residing in the gastrointestinal tract, plays a crucial role in maintaining human health and well-being. This microbial community is not just a passive bystander but actively contributes to numerous physiological processes essential for human health including mental fitness, immunological and metabolic mechanisms.

引言肠道健康最近受到了相当大的关注，并已成为大多数健身制度的重要组成部分。肠道微生物群由居住在胃肠道中的数万亿微生物组成，在维持人类健康和福祉方面起着至关重要的作用。这种微生物群落不仅是一个被动的旁观者，而且积极地促进了许多对人类健康至关重要的生理过程，包括心理健康，免疫和代谢机制。

However, this niche of vital microbes gets easily disrupted due to several factors including the consumption of foods lacking fibre content, lack of exercise, stress, excessive intake of antibiotics, proton pump inhibitors (PPIs), and non-steroidal anti-inflammatory drugs (NSAIDs), etc. Among these factors affecting the gut microbiome, the food we consume plays a major role in regulating our health1.The healthy food items or supplements we consume undergo several physical and chemical changes during digestion in our gastrointestinal tract before the nutrients from them get absorbed into our body.

然而，由于多种因素，包括食用缺乏纤维含量的食物，缺乏运动，压力，过量摄入抗生素，质子泵抑制剂（PPIs）和非甾体抗炎药（NSAIDs）等，这种重要微生物的生态位很容易被破坏。在这些影响肠道微生物组的因素中，我们食用的食物在调节我们的健康方面起着重要作用1。我们食用的健康食品或补充剂在消化过程中会发生一些物理和化学变化，然后营养物质就会被吸收到我们的体内。

These nutrients or bioactives present in our vegetables, herbs, spices, or fruits serve as cofactors, vitamins, or essential factors for better functioning of our body. However, their absorption in the small intestine is often affected by several factors including chemical form, matrix, and metabolic pathways2.

这些存在于我们的蔬菜、草药、香料或水果中的营养素或生物活性物质可以作为辅助因子、维生素或人体更好功能的必需因素。然而，它们在小肠中的吸收通常受到多种因素的影响，包括化学形式，基质和代谢途径2。

Therefore, it is essential to develop a suitable delivery matrix to ensure the bioavailability of essential bioactives for a healthy population. Advances in nanotechnology offer us vast opportunities to manipulate the chemical, physical and biological properties of various compounds to suit our goals.

因此，必须开发合适的递送基质，以确保健康人群必需生物活性的生物利用度。纳米技术的进步为我们提供了巨大的机会来操纵各种化合物的化学，物理和生物学特性，以满足我们的目标。

One such development is the synthesis of nano-delivery systems made with bioco.

一个这样的发展是用bioco制成的纳米递送系统的合成。

Male flies post-exposure to respective treatment groups were homogenised in suitable buffers to estimate redox stress parameters. Following the behaviour analysis, baseline control groups (CS, CS NPs and probiotics) along with control, ACR and CSPA groups were tested for fluctuations in the ROS levels.

将暴露于各个处理组后的雄性苍蝇在合适的缓冲液中匀浆以估计氧化还原应激参数。在行为分析之后，测试基线对照组（CS，CS NPs和益生菌）以及对照组，ACR和CSPA组的ROS水平波动。

As illustrated in Fig. 6A, there were no significant variations in ROS levels between the control and baseline control groups. The observed ROS levels are commonly attributed to normal cellular activity, however, a marked increase in ROS intensity was noticed in flies treated with ACR thereby indicating the presence of xenobiotic-induced oxidative stress and activation of antioxidant mechanisms.

如图6A所示，对照组和基线对照组之间的ROS水平没有显着变化。观察到的ROS水平通常归因于正常的细胞活性，然而，在用ACR处理的果蝇中注意到ROS强度显着增加，从而表明存在异生素诱导的氧化应激和抗氧化机制的激活。

Owing to the non-toxic nature of the baseline control groups (CS, CS NPs and probiotics) as evidenced by long survival period, high locomotor function at larval and fly stages and no significant induction of ROS levels, further experiments were performed with control, ACR, CSP NPs and CSPA treatment groups.Fig.

由于基线对照组（CS，CS-NPs和益生菌）的无毒性，如存活期长，幼虫和苍蝇阶段的高运动功能以及ROS水平没有显着诱导所证明的，进一步的实验用对照，ACR，CSP-NPs和CSPA治疗组进行。图。

6Biochemical Factors of D.mel. The effects of CSP NP were tested against ACR-induced ROS levels (A), antioxidant enzyme activity (SOD—B and GST—C) and protein levels (D) using the fruit fly model.Full size imageACR-treated flies exhibited increased ROS, SOD and GST activities as depicted in Fig. 6A–C respectively.

6 D.mel的生物化学因子。使用果蝇模型测试CSP-NP对ACR诱导的ROS水平（A），抗氧化酶活性（SOD-B和GST-C）和蛋白质水平（D）的影响。。

Furthermore, ACR exposure indicated a negative influence on protein levels (Fig. 6D). On the other hand, the CSPA-treated group demonstrated a significant recovery in ROS and antioxidant enzyme activity levels. Comparing the data with the control group revealed the prominent effect of CSPA treatment in restoring the alterations in vital biochemical factors and thereby curbing the toxic influence of ACR within the fruit fly model.CSP NPs reverse ACR-induced mitocho.

此外，ACR暴露表明对蛋白质水平有负面影响（图6D）。另一方面，CSPA治疗组表现出ROS和抗氧化酶活性水平的显着恢复。将数据与对照组进行比较，发现CSPA治疗在恢复重要生化因子的改变方面具有显着效果，从而抑制了果蝇模型中ACR的毒性影响。CSP-NPs逆转ACR诱导的线粒体。

Drosophila rearing and maintenanceThe Drosophila Oregon K wild-type strain was maintained at 25 ± 2 °C and 60% humidity under a 12 h dark–light cycle. Flies were cultured on a standard cornmeal agar medium consisting of cornflour, agar–agar type 1, D-glucose, sugar, and yeast extract. To prevent microbial contamination, the medium was autoclaved and supplemented with antifungal agents including propionic acid, Tego (methyl para hydroxy benzoate dissolved in ethanol), and orthophosphoric acid at 55 °C.

果蝇饲养和维护果蝇俄勒冈州K野生型菌株在12小时的黑暗-光照循环下保持在25±2°C和60%的湿度。。为了防止微生物污染，将培养基高压灭菌并在55℃下补充抗真菌剂，包括丙酸，Tego（溶于乙醇的对羟基苯甲酸甲酯）和正磷酸。

As baseline control groups—0.5% chitosan (CS), 10 µg/ml CS NPs and 3 ml of L. fermentum culture (8 log CFU/ml) treatment groups were analysed to comprehend the impact on behaviour and ROS levels. Additionally, 2 mM acrylamide (ACR), 10 µg/ml CSP NPs, and 10 µg/ml CSP NPs + ACR (CSPA) treatment groups were prepared by the stoichiometric addition of respective compounds to the media at 50–55 °C.Survival and behavioural assayThe lifespan of flies was estimated by transferring twenty-five healthy newborn adult male flies to freshly prepared treatment media.

作为基线对照组，分析了0.5%壳聚糖（CS），10µg/ml CS NPs和3 ml发酵乳杆菌培养物（8 log CFU/ml）处理组，以了解对行为和ROS水平的影响。此外，通过在50-55°C下向培养基中化学计量添加相应化合物，制备了2 mM丙烯酰胺（ACR），10µg/ml CSP NPs和10µg/ml CSP NPs+ ACR（CSPA）处理组。存活和行为测定通过将25只健康的新生成年雄性苍蝇转移到新鲜制备的治疗培养基中来估计苍蝇的寿命。

The flies were then constantly monitored and the mortality rate was calculated by tallying the number of dead flies every 24 h. To assess the locomotor functions at the larval stage, male and female flies were added to the respective media for 5 days. The parent flies were then discarded and the vials were carefully maintained till the development of third instar larvae.

然后不断监测苍蝇，并通过每24小时统计死蝇数量来计算死亡率。为了评估幼虫阶段的运动功能，将雄性和雌性苍蝇添加到各自的培养基中5天。。

These larvae were then transferred to an agar plate placed on top of a graph paper and allowed to crawl for one minute. The number of 1 mm grids traversed per minute was represented as the crawling activity for statistical analysis. Similarly, the adult climbing activity was determined by exposing thirty male flies to the respective treatment media for 5 da.

然后将这些幼虫转移到放在方格纸上的琼脂平板上，让其爬行一分钟。每分钟遍历的1毫米网格数表示为用于统计分析的爬行活动。类似地，通过将30只雄性苍蝇暴露于各自的处理介质中5 da来确定成年攀爬活动。

Data availability

数据可用性

All data sets used and/or generated in this work are obtainable from the corresponding author upon reasonable request.

在这项工作中使用和/或生成的所有数据集都可以根据合理的要求从相应的作者那里获得。

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Download referencesAcknowledgementsThe authors acknowledge the SRM SCIF and DBT platform for Advanced Life Science’s support in TEM, fluorescence imaging studies and Zeta analysis. The authors also recognize the support provided by the Department of Science and Technology, Ministry of Science and Technology, Government of India through the INSPIRE Fellowship program.Author informationAuthors and AffiliationsDevelopmental Biology Lab, Department of Biotechnology, School of Bioengineering, SRM Institute of Science and Technology, Kattankulathur, 603203, Tamil Nadu, IndiaSwetha Senthil Kumar & Sahabudeen Sheik MohideenAuthorsSwetha Senthil KumarView author publicationsYou can also search for this author in.

下载参考文献致谢作者感谢SRM SCIF和DBT平台在TEM，荧光成像研究和Zeta分析方面对高级生命科学的支持。作者还感谢印度政府科学技术部科学技术司通过INSPIRE奖学金计划提供的支持。作者信息作者和附属机构SRM科学与技术研究所生物工程学院生物技术系发展生物学实验室，Kattankulathur，603203，泰米尔纳德邦，IndiaSwetha Senthil Kumar＆Sahabudeen Sheik MohideenAuthorsSwetha Senthil KumarView作者出版物您也可以在中搜索这位作者。

PubMed Google ScholarSahabudeen Sheik MohideenView author publicationsYou can also search for this author in

PubMed谷歌学者Sahabudeen Sheik MohideenView作者出版物您也可以在

PubMed Google ScholarContributionsS.S.—Conceptualization, Methodology, Data curation, Writing—original draft, Writing—review and editing, Software, Validation; S.S.M.—Conceptualization, Methodology, Validation, Writing—review and editing and Supervision.Corresponding authorCorrespondence to.

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Reprints and permissionsAbout this articleCite this articleSenthil Kumar, S., Sheik Mohideen, S. Chitosan-coated probiotic nanoparticles mitigate acrylamide-induced toxicity in the Drosophila model.

转载和许可本文引用本文Senthil Kumar，S.，Sheik Mohideen，S。壳聚糖包被的益生菌纳米粒子减轻了果蝇模型中丙烯酰胺诱导的毒性。

Sci Rep 14, 21182 (2024). https://doi.org/10.1038/s41598-024-72200-wDownload citationReceived: 22 July 2024Accepted: 04 September 2024Published: 11 September 2024DOI: https://doi.org/10.1038/s41598-024-72200-wShare this articleAnyone you share the following link with will be able to read this content:Get shareable linkSorry, a shareable link is not currently available for this article.Copy to clipboard.

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KeywordsGut microbiotaNanoencapsulationAcrylamideBioactivesChitosan

关键词：微生物群无包膜丙烯酰胺生物壳聚糖

Lactobacillus

乳酸杆菌

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