Abstract
Diabetes is a major global health concern. This study aimed to investigate the correlation between differentially expressed lncRNAs in mice with type 2 diabetes mellitus (T2DM) and alterations in the intestinal flora and intestinal pathology. A T2DM mouse model was constructed by feeding mice a high-fat diet. Serum fat metabolism-related indices and insulin levels were biochemically detected. Serum inflammatory factors (IL-1β, IL-6, TNF-α, IL-10) and endotoxin (LPS) were measured by ELISA. Histopathological changes in the small intestines of mice were observed by HE. The short-chain fatty acid (SCFA) content was analyzed using GC-MS. Analysis of altered intestinal flora in T2DM mice was performed using a 16sRNA sequencing assay. Differences in lncRNA expression profiles in small intestinal tissues were analyzed using RNA-seq assays. Spearman’s correlation analysis was used to correlate the expression of candidate lncRNAs with changes in differential gut flora. Spearman’s correlation analysis was used to analyze the correlation between the expression of candidate differentially expressed lncRNAs, small intestinal permeability, and glucose absorption. We found that serum levels of LPS, BUN, Scr, TC, TG, LDL-C, IL-1β, IL-6, and TNF-α were elevated and levels of HDL-C, insulin, and IL-10 were decreased in T2DM mice. The ileal enterochromes of T2DM mice were disorganized and broken, the number of enterochromes was reduced, the local epithelial cells were necrotic, and the plasma membrane layer was locally absent. In addition, the protein expression of ZO-1 and occludin was decreased, and the protein expression of SGLT-1 and GLUT-2 was elevated in the model group compared to the control group. The levels of Acetic acid, Propionic acid and Butyric acid were decreased and the levels of Isobutyric acid and Isovaleric acid were increased, the abundance of beneficial bacteria was decreased and the abundance of harmful bacteria was increased in the feces of T2DM mice. RNA-seq identified nine differentially expressed lncRNAs (LINC00675, Gm33838, Gm11655, LOC6613926, LOC6613788, LOC6613791, LOC6613795, Arhgap27os3, and A330023F24Rik). In addition, we found significant correlations between differentially expressed lncRNAs and a variety of intestinal flora, as well as between small intestinal permeability and glucose absorption. A significant correlation was observed between differentially expressed lncRNAs in the intestinal tissues of T2DM mice and intestinal flora imbalance, small intestinal permeability, and glucose absorption.
Similar content being viewed by others
Data Availability
The data used to support the findings of this study have been included in this article.
References
Yang, Q., Vijayakumar, A., & Kahn, B. B. (2018). Metabolites as regulators of insulin sensitivity and metabolism. Nature Reviews Molecular Cell Biology, 19(10), 654–672.
Chatterjee, S., Khunti, K., & Davies, M. J. (2017). Type 2 diabetes. Lancet, 389(10085), 2239–2251.
Gilbert, J. A., Blaser, M. J., Caporaso, J. G., Jansson, J. K., Lynch, S. V., & Knight, R. (2018). Current understanding of the human microbiome. Nature Medicine, 24(4), 392–400.
Burcelin, R., Serino, M., Chabo, C., Blasco-Baque, V., & Amar, J. (2011). Gut microbiota and diabetes: From pathogenesis to therapeutic perspective. Acta Diabetologica, 48(4), 257–273.
Ley, R. E., Hamady, M., Lozupone, C., Turnbaugh, P. J., Ramey, R. R., Bircher, J. S., Schlegel, M. L., Tucker, T. A., Schrenzel, M. D., Knight, R., & Gordon, J. I. (2008). Evolution of mammals and their gut microbes. Science, 320(5883), 1647–1651.
Zhang, C., Zhang, M., Wang, S., Han, R., Cao, Y., Hua, W., Mao, Y., Zhang, X., Pang, X., Wei, C., Zhao, G., Chen, Y., & Zhao, L. (2010). Interactions between gut microbiota, host genetics and diet relevant to development of metabolic syndromes in mice. The ISME Journal, 4(2), 232–241.
Qin, J., Li, Y., Cai, Z., Li, S., Zhu, J., Zhang, F., Liang, S., Zhang, W., Guan, Y., Shen, D., Peng, Y., Zhang, D., Jie, Z., Wu, W., Qin, Y., Xue, W., Li, J., Han, L., Lu, D., Wu, P., Dai, Y., Sun, X., Li, Z., Tang, A., Zhong, S., Li, X., Chen, W., Xu, R., Wang, M., Feng, Q., Gong, M., Yu, J., Zhang, Y., Zhang, M., Hansen, T., Sanchez, G., Raes, J., Falony, G., Okuda, S., Almeida, M., LeChatelier, E., Renault, P., Pons, N., Batto, J. M., Zhang, Z., Chen, H., Yang, R., Zheng, W., Li, S., Yang, H., Wang, J., Ehrlich, S. D., Nielsen, R., Pedersen, O., Kristiansen, K., Wang, J. (2012). A metagenome-wide association study of gut microbiota in type 2 diabetes. Nature, 490(7418), 55-60
Yano, J. M., Yu, K., Donaldson, G. P., Shastri, G. G., Ann, P., Ma, L., Nagler, C. R., Ismagilov, R. F., Mazmanian, S. K., & Hsiao, E. Y. (2015). Indigenous bacteria from the gut microbiota regulate host serotonin biosynthesis. Cell, 161(2), 264–276.
Ma, Q., Li, Y., Li, P., Wang, M., Wang, J., Tang, Z., Wang, T., Luo, L., Wang, C., Wang, T., & Zhao, B. (2019). Research progress in the relationship between type 2 diabetes mellitus and intestinal flora. Biomedicine & Pharmacotherapy, 117, 109138.
Gurung, M., Li, Z., You, H., Rodrigues, R., Jump, D. B., Morgun, A., & Shulzhenko, N. (2020). Role of gut microbiota in type 2 diabetes pathophysiology. EBioMedicine., 51, 102590.
Harsch, I. A., Konturek, P. C. (2018). The role of gut microbiota in obesity and type 2 and type 1 diabetes mellitus: New insights into “old” diseases. Medical Sciences (Basel), 6(2), 32.
Yang, F., Chen, Y., Xue, Z., Lv, Y., Shen, L., Li, K., Zheng, P., Pan, P., Feng, T., Jin, L., & Yao, Y. (2020). High-throughput sequencing and exploration of the lncRNA-circRNA-miRNA-mRNA network in type 2 diabetes mellitus. BioMed Research International, 2020, 8162524.
Yang, Y. L., Xue, M., Jia, Y. J., Hu, F., Zheng, Z. J., Wang, L., Si, Z. K., & Xue, Y. M. (2020). Long noncoding RNA NEAT1 is involved in the protective effect of Klotho on renal tubular epithelial cells in diabetic kidney disease through the ERK1/2 signaling pathway. Experimental & Molecular Medicine, 52(2), 266–280.
Yu, C., Yang, K., Meng, X., Cao, B., & Wang, F. (2020). Downregulation of long noncoding RNA MIAT in the retina of diabetic rats with tail-vein injection of human umbilical-cord mesenchymal stem cells. International Journal of Medical Sciences, 17(5), 591–598.
Zhang, L., & Wang, Y. M. (2019). Expression and function of lncRNA ANRIL in a mouse model of acute myocardial infarction combined with type 2 diabetes mellitus. Journal of the Chinese Medical Association, 82(9), 685–692.
Zhang, W., Zheng, J., Hu, X., & Chen, L. (2019). Dysregulated expression of long noncoding RNAs serves as diagnostic biomarkers of type 2 diabetes mellitus. Endocrine, 65(3), 494–503.
Brunkwall, L., & Orho-Melander, M. (2017). The gut microbiome as a target for prevention and treatment of hyperglycaemia in type 2 diabetes: From current human evidence to future possibilities. Diabetologia, 60(6), 943–951.
Bäckhed, F. (2012). Host responses to the human microbiome. Nutrition Reviews, 70(Suppl 1), S14-17.
Zhang, Q., & Hu, N. (2020). Effects of metformin on the gut microbiota in obesity and type 2 diabetes mellitus. Diabetes, Metabolic Syndrome and Obesity: Targets and Therapy, 13, 5003–5014.
Candela, M., Biagi, E., Soverini, M., Consolandi, C., Quercia, S., Severgnini, M., Peano, C., Turroni, S., Rampelli, S., Pozzilli, P., Pianesi, M., Fallucca, F., & Brigidi, P. (2016). Modulation of gut microbiota dysbioses in type 2 diabetic patients by macrobiotic Ma-Pi 2 diet. British Journal of Nutrition, 116(1), 80–93.
Zhang, X., Shen, D., Fang, Z., Jie, Z., Qiu, X., Zhang, C., Chen, Y., & Ji, L. (2013). Human gut microbiota changes reveal the progression of glucose intolerance. PLoS One, 8(8), e71108.
Lippert, K., Kedenko, L., Antonielli, L., Kedenko, I., Gemeier, C., Leitner, M., Kautzky-Willer, A., Paulweber, B., & Hackl, E. (2017). Gut microbiota dysbiosis associated with glucose metabolism disorders and the metabolic syndrome in older adults. Beneficial Microbes, 8(4), 545–556.
Aw, W., & Fukuda, S. (2018). Understanding the role of the gut ecosystem in diabetes mellitus. Journal of Diabetes Investigation, 9(1), 5–12.
Yan, F., Li, N., Shi, J., Li, H., Yue, Y., Jiao, W., Wang, N., Song, Y., Huo, G., & Li, B. (2019). Lactobacillus acidophilus alleviates type 2 diabetes by regulating hepatic glucose, lipid metabolism and gut microbiota in mice. Food & Function, 10(9), 5804–5815.
Qamar, H., Hussain, K., Soni, A., Khan, A., Hussain, T., Chénais, B. (2021). Cyanobacteria as natural therapeutics and pharmaceutical potential: Role in antitumor activity and as nanovectors. Molecules, 26(1), 247.
Plovier, H., Everard, A., Druart, C., Depommier, C., Van Hul. M., Geurts, L., Chilloux, J., Ottman, N., Duparc, T., Lichtenstein, L., Myridakis, A., Delzenne, N. M., Klievink, J,, Bhattacharjee, A., van der Ark, K. C., Aalvink, S., Martinez, L. O., Dumas, M. E., Maiter, D., Loumaye, A., Hermans, M. P., Thissen, J. P., Belzer, C., de Vos W. M., Cani, P. D. (2017). A purified membrane protein from Akkermansia muciniphila or the pasteurized bacterium improves metabolism in obese and diabetic mice. Nature Medicine, 23(1), 107-113
Shen, Z., Zhu, C., Quan, Y., Yang, J., Yuan, W., Yang, Z., Wu, S., Luo, W., Tan, B., & Wang, X. (2018). Insights into Roseburia intestinalis which alleviates experimental colitis pathology by inducing anti-inflammatory responses. Journal of Gastroenterology and Hepatology, 33(10), 1751–1760.
Rachdaoui, N. (2020). Insulin: The friend and the foe in the development of type 2 diabetes mellitus. International Journal of Molecular Sciences, 21(5), 1770.
Cani, P. D., Amar, J., Iglesias, M. A., Poggi, M., Knauf, C., Bastelica, D., Neyrinck, A. M., Fava, F., Tuohy, K. M., Chabo, C., Waget, A., Delmée, E., Cousin, B., Sulpice, T., Chamontin, B., Ferrières, J., Tanti, J. F., Gibson, G. R., Casteilla, L. ,…, Burcelin, R. (2007). Metabolic endotoxemia initiates obesity and insulin resistance. Diabetes, 56(7), 1761–1772.
Kim, C. H. (2018). Microbiota or short-chain fatty acids: Which regulates diabetes? Cellular & Molecular Immunology, 15(2), 88–91.
Puddu, A., Sanguineti, R., Montecucco, F., & Viviani, G. L. (2014). Evidence for the gut microbiota short-chain fatty acids as key pathophysiological molecules improving diabetes. Mediators of Inflammation, 2014, 162021.
Lassenius, M. I., Pietiläinen, K. H., Kaartinen, K., Pussinen, P. J., Syrjänen, J., Forsblom, C., Pörsti, I., Rissanen, A., Kaprio, J., Mustonen, J., Groop, P. H., & Lehto, M. (2011). Bacterial endotoxin activity in human serum is associated with dyslipidemia, insulin resistance, obesity, and chronic inflammation. Diabetes Care, 34(8), 1809–1815.
Pussinen, P. J., Havulinna, A. S., Lehto, M., Sundvall, J., & Salomaa, V. (2011). Endotoxemia is associated with an increased risk of incident diabetes. Diabetes Care, 34(2), 392–397.
Yoshida, N., Emoto, T., Yamashita, T., Watanabe, H., Hayashi, T., Tabata, T., Hoshi, N., Hatano, N., Ozawa, G., Sasaki, N., Mizoguchi, T., Amin, H. Z., Hirota, Y., Ogawa, W., Yamada, T., & Hirata, K. I. (2018). Bacteroides vulgatus and Bacteroides dorei reduce gut microbial lipopolysaccharide production and inhibit atherosclerosis. Circulation, 138(22), 2486–2498.
Ruan, Y., Lin, N., Ma, Q., Chen, R., Zhang, Z., Wen, W., Chen, H., & Sun, J. (2018). Circulating lncRNAs analysis in patients with type 2 diabetes reveals novel genes influencing glucose metabolism and islet β-cell function. Cellular Physiology and Biochemistry, 46(1), 335–350.
Li, L., Li, C., Lv, M., Hu, Q., Guo, L., & Xiong, D. (2020). Correlation between alterations of gut microbiota and miR-122-5p expression in patients with type 2 diabetes mellitus. Annals of Translational Medicine, 8(22), 1481.
Funding
This project was supported by the Natural Science Foundation of China (grant no. 81500427) and the Angel Nutritech Nutrition Fund (AF2017002).
Author information
Authors and Affiliations
Contributions
Shufang-Xu prepared the manuscript and provided the experimental guidance. Guidance on writing experimental research papers was provided by Heng-Zhang and Hui-Zou. The experimental research and data collection were conducted by Zhitao-Chen, Ting-Jiang, and Mengjun-Huang.
Corresponding author
Ethics declarations
Ethical Approval
The experiment was approved by the Animal Ethics Committee of Wuhan Myhalic Biotechnology Co., Ltd (HLK-20210621-001).
Consent to Participate
All authors participated in the study.
Consent for Publication
All authors agree to publish this article.
Competing Interests
The authors declare no competing interests.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Xu, S., Zhang, H., Zou, H. et al. Correlation of Differentially Expressed lncRNAs with Intestinal Flora Imbalance, Small Intestinal Permeability, and Glucose Uptake in T2DM Mice. Appl Biochem Biotechnol (2024). https://doi.org/10.1007/s12010-024-04935-1
Accepted:
Published:
DOI: https://doi.org/10.1007/s12010-024-04935-1