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    Home > Food News > Food Articles > Effects of J. Future Foods Rhodiola rosea extract on insulin resistance in Caenorhabditis elegans

    Effects of J. Future Foods Rhodiola rosea extract on insulin resistance in Caenorhabditis elegans

    • Last Update: 2022-11-14
    • Source: Internet
    • Author: User
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    Insulin resistance refers to a pathological condition
    in which the body's sensitivity to insulin decreases, leading to glucose uptake and utilization affected.
    Rhodiola rosea L.
    is a perennial herb widely distributed in Tibet, Qinghai, Yunnan and other alpine regions of China, and Rhodiola rosea and salidroside are likely to reduce blood sugar levels through the regulation of the AMPK signaling pathway, thereby improving diabetes and its complications
    Caenorhabditis elegans is a powerful model organism in diabetes research, characterized by its ease of use, relatively short lifespan, high operability and low cost

    In this paper, Professor Teng Hui and Chen Lei of the College of Food Science and Technology of Guangdong Ocean University used Rhodiola rosea as a model to explore the effects of Rhodiola rosea extract intervention on oxidase, reactive oxygen species accumulation level and apoptosis in high-sugar cultured Caenorhabditis elegans, so as to evaluate the insulin resistance effect
    of Rhodiola rosea extract EAE on Caenorhabditis elegans 。 The effect of Rhodiola rosea extract on the expression levels of DAF-2, DAF-16, AKT-1, SKN-1, AAK-2 and SOD-3 genes was determined by q-PCR technology, and the molecular mechanism
    of Rhodiola rosea extract on insulin resistance in Caenorhabditis elegans was determined.

    Results and discussion

    Insulin signaling analysis

    1 Adult rate of Caenorhabditis elegans after intervention

    DR1564 adult worm and dauer worm appearance difference is obvious, as shown in Figure 1, the adult body is fatter, the head and tail are rounded, and the eggs can be seen under the microscope; The dauer stage nematodes have a slender body and fast swimming speed, which can be quickly distinguished under the microscope

    After 120 h of rescue treatment, there was no significant difference in the adult rate between the blank group and the high-glucose model group, indicating that the high glucose culture did not affect the effect
    of improving insulin strength.
    Compared with the blank group, the worm rates of metformin and sodium vanadate in the low- and high-dose EAE group were significantly different from those in the blank group, and the treatment effect was best in the high-dose EAEH group, and the adult worm rate increased by 52.
    compared with the blank group.
    This indicates that after 120 hours of drug intervention, more than half of Caenorhabditis elegans were rescued from the dauer stage, and the intervention effect of the drug was positively correlated with
    the dose administered.

    Glucose level measurement

    2 Glucose content in Caenorhabditis elegans

    Caenorhabditis elegans significantly increased glucose levels in vitro under NGM medium with a glucose concentration of 40 mmol/L, indicating that glucose addition greatly increased glucose content
    in nematodes.
    After the administration intervention, it can be clearly observed that each group shows different degrees of hypoglycemic effects, the effect of low-dose EAEL group is basically the same as that of metformin group, and sodium vanadate also shows a good hypoglycemic effect, but they fail to restore the glucose content to normal levels
    The high-dose EAEH group had the strongest ability to reduce glucose and restore glucose levels to normal levels

    Reactive oxygen species (ROS) level determination

    3 Effects on ROS levels in Caenorhabditis elegans in a high-glucose environment led to the accumulation of reactive oxygen species levels in nematodes, and the ROS level in the model group was 1.
    71 times
    that of the blank group.
    After treatment with EAEL, EAEH, metformin, and sodium vanadate, ROS levels decreased significantly, but did not show dose-dependence

    Antioxidant enzyme (SOD, CAT) levels are measured

    4 Effects on the levels of SOD(A), CAT(B), GSH-Px(C), GSH(D) in Caenorhabditis elegans

    The activities of superoxide dismutase (SOD) and catalase (CAT) in Caenorhabditis elegans cultured in high sugar environment were significantly reduced in model group, EAEL group, EAEH group, metformin group and sodium vanadate group, and did not play a good free radical scavenging effect
    From this, it can be inferred that one of the possible ways in which ethyl Rhodiola acetate extract reduces ROS levels in nematodes is by increased superoxide
    dismutase activity and catalase activity.

    GSH-Px enzyme and GSH content determination

    Glutathione (GSH) is a small molecule peptide composed of three amino acid molecules with detoxification effect, which can bind to free radicals, turn harmful substances into harmless and excrete them from the body
    Glutathione peroxidase (GSH-Px), as a marker of oxidative stress, can catalyze the conversion of GSH to GSSH, and is an important peroxidase
    widely found in the body.
    In Figure 6, the activity of GSH-Px decreased significantly under high sugar environment, and the GSH-Px level after administration intervention was significantly increased compared with the model group, and the reduced GSH content increased
    The decrease of GSH-Px enzyme activity leads to GSH can not be catalyzed in time, and its content shows a clear upward trend, and the decrease of glutathione peroxidase activity and the increase of glutathione content are also important factors
    leading to the accumulation of reactive oxygen species in nematodes.

    elegans cell apoptosis

    Apoptosis is an evolutionarily conserved cellular self-destruction program that requires ensuring functional organ structure and maintaining tissue
    In Caenorhabditis elegans, acridine orange stain binds to DNA through cell molds and fluoresces
    green under the microscope.
    In apoptotic cells, DNA is broken up and divided into fragments of different sizes, also called apoptotic bodies, and acridine orange can make these fragments appear dense yellow-green fluorescence under a fluorescence
    Compared with the blank group, the relative fluorescence intensity of each group was significantly different, indicating that oxidative stress caused by high glucose culture greatly promoted apoptosis
    Compared with the model group, apoptosis was delayed to a certain extent after 120 h of administration, but it was not obvious, and only the high-dose EAE group had significant differences, showing a certain protective effect
    of apoptosis.

    5 Effect on apoptosis in Caenorhabditis elegans cells

    DAF-2 and DAF-16 gene expression levels

    6 Effects on gene expression in Caenorhabditis elegans DAF-2(A) and DAF-16(B).

    The insulin/IGF-1 signaling pathway (IIS) pathway controls aging in nematodes, which play a vital role
    in their lifespan.
    In the IIS pathway, DAF-2 and DAF-16 are widely recognized as key genes
    that affect the lifespan of nematodes.
    Compared with the model group, the expression of DAF-2 and DAF-16 genes was upregulated
    to a certain extent.
    Among them, EAEH, metformin and sodium vanadate showed obvious upregulation
    of DAF-2 gene expression.
    There was no clear difference in
    DAF-16 expression after dosing intervention compared to the model group.
    Only the EAEH and metformin groups showed a non-significant upward adjustment trend
    It can be speculated that Rhodiola rosea may have the effect
    of prolonging life by upregulating the expression of DAF-2 and DAF-16 genes.
    And it plays a significant dose-dependent role in the regulation of DAF-2

    AKT-1 and SKN-1 gene expression levels

    SKN-1 transcription factor is an evolutionarily conserved exogenous stress regulator and longevity factor that alleviates oxidative stress damage by regulating the expression of antioxidant enzyme-related genes
    AKT is an important factor affecting the function of islet β cells and insulin signaling, which is closely related
    to glucose uptake and metabolism.
    In the gene expression of AKT-1 and SKN-1, there was no difference between the model group and the blank group, the low-dose EAEL group showed significant downregulation, and high-dose EAE, metformin, and sodium vanadate were significantly upregulated
    Therefore, it is speculated that Rhodiola rosea can improve insulin signaling and promote energy metabolism by upregulating SKN-1 and AKT-1 expressions, so as to improve insulin resistance

    7 Effects on the expression of AKT-1(A) and SKN-1(B) genes in Caenorhabdititis elegans

    AAK-2 and SOD-3 gene expression levels

    8 Effects on AAK-2 and SOD-3 gene expression in Caenorhabditis elegans

    AAK-2 is a Caenorhabditis elegans homolog of the subunit α AMP-activated protein kinase (AMPK) that regulates cellular energy homeostasis and plays a role
    in fighting stress and prolonging life.
    SOD-3, as a member of the superoxide dismutase family in C.
    elegans, is associated
    with oxidative damage in nematodes.
    EAEL, EAEH, and metformin significantly upregulated the expression
    of AAK-2.
    High sugar induced oxidative stress led to a general increase in SOD-3 expression, and SOD-3 was significantly downregulated
    in the low-dose EAEL group and sodium vanadate group after the administration intervention.
    This suggests that Rhodiola rosea does not improve insulin resistance by upregulating SOD-3 expression, but may play a protective role
    against insulin resistance by increasing AAK-2 gene expression.


    Rhodiola rosea extract had a certain effect on the T2DM model of DR1564 nematode, increasing its insulin signaling and significantly reducing the glucose level in the nematode to a close to the fasting blood glucose value
    of a person.
    After 120 h of administration of Rhodiola rosea extract, it can significantly reduce the ROS level in nematodes, significantly increase the activity of GSH-Px, and reduce the activity of GSH-Px enzyme, resulting in the failure of GSH to be converted in time and a large amount of accumulation
    in nematodes 。 In addition, Rhodiola rosea extract can reduce the apoptosis rate of nematode cells under a high-sugar diet, and can also activate related pathways to strengthen insulin signaling, promote nutrient absorption and metabolism, enhance antioxidant capacity by upregulating the expression of DAF-2, DAF-16, AKT-1, SKN-1, and AAK-2 genes, and ultimately improve the insulin resistance of C.

    About the first author

    Teng Hui, female, Ph.
    in Food Science and Biotechnology, Kyungpook University, South Korea, associate professor and master supervisor
    of Guangdong Ocean University.
    Main research contents: efficient extraction of natural antioxidants in food and research and development of functional foods; Control of hazards and changes in nutritional composition during food processing; Rapid qualitative quantification, separation and purification of bioactive ingredients in food, and the use of cell models and animal models to evaluate their bioavailability (hypoglycemia, hypolipidemia, anti-inflammation, prevention of oxidative damage, etc.
    ), to establish the structure-activity relationship
    between active ingredients and bioavailability.
    He has published more than 30 SCI papers as the first author or corresponding author

    About the corresponding author

    Chen Lei, male, Ph.
    in Agricultural Life Sciences, Kyungpook University, South Korea, professor and doctoral supervisor of Guangdong Ocean University, high-level introduced talents of Guangdong Ocean University in 2021, winner of Fujian Agriculture and Forestry University's
    "School Outstanding Youth" 。 Associate Editor of eFood, Editorial Board Member of Frontiers in Pharmacology and Food Science and Human Wellness, Oxidative Medicine and Cellular Longevity, Frontiers in Pharmacology and Canadian Journal of Gastroenterology and Guest Editor of the special issue of Hepatology, Secretary of the International Association of dietetic nutrition and safety (IADNS).

    。 He has presided over 5 projects such as the National Natural Science Foundation of China, the Youth Fund, and the China Postdoctoral Program, and has published more than 70 SCI/EI papers as the first or corresponding author, including 30 papers in the first region, 10 papers in IF>10, 6 highly cited papers, 1 hot paper, and the total number of papers cited > 1500 times (Elsevier), H-index=25
    His main research areas include phytochemicals, bioavailability of functional nutrients

    Effects of Rhodiola rosea and its major compounds on insulin resistance in Caenorhabditis elegans

    Hui Tenga, Hongting Denga, Yanzi Wub, Chang Zhanga, Chao Aia, Hui Caoa, Jianbo Xiaoa, Lei Chena,*

    a Guangdong Provincial Key Laboratory of Aquatic Product Processing and Safety, Guangdong Province Engineering Laboratory for Marine Biological Products, Guangdong Provincial Engineering Technology Research Center of Seafood, Key Laboratory of Advanced Processing of Aquatic Product of Guangdong Higher Education Institution, College of Food Science and Technology, Guangdong Ocean University, Zhanjiang 524088, China

    b College of Food Science, Fujian Agriculture and Forestry University, Zhuzhou 350002, China

    *Corresponding author.

    E-mail address: chenlei841114@hotmail.


    By establishing insulin resistance model of Caenorhabditis elegans, the effects of Rhodiola rosea extract and its compounds on glucose level, insulin signal intensity, antioxidant enzymes and cell apoptosis of C.
    elegans were investigated in present study.
    The mRNA expression of DAF-2, DAF-16, AkT-1, SKN-1, AAK-2 and SOD-3 were detected by qRT-PCR.
    The results revealed that R.
    rosea extract contain salidroside, kaempferol, chlorogenic acid, kaemphenol-7-O-glucoside, caffeic acid, and quercetin, etc.
    At the concentration of 200 μg/mL, R.
    rosea extract treated C.
    elegans showed increased insulin signal intensity by 52.
    46% compared with the normal group, while, glucose content decreased by 72.
    13% and reactive oxygen species (ROS) level decreased by 36.
    Compared with high glucose model group, the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) increased by 10.
    9, 14.
    2 and 27.
    9, respectively.
    After R.
    rosea extract intervention, the expression levels of DAF-2, AkT-1, AAK-2, SKN-1 and other genes significantly increased.
    These results also indicated that R.
    rosea extract could activate insulin receptor substrate (IRS) and adenosine 5′-monophosphate (AMP)-activated protein kinase (AMPK) pathways to enhance insulin signaling, promote nutrient metabolism, and achieve the effect for improving insulin resistance of C.


    TENG H, DENG H T, WU Y Z, et al.
    Effects of Rhodiola rosea and its major compounds on insulin resistance in Caenorhabditis elegans[J].
    Journal of Future Foods, 2022, 2(4): 365-371.

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