The transcriptomic-based disease network reveals synergistic therapeutic effect of total alkaloids from Coptis chinensis and total ginsenosides from Panax ginseng on type 2 diabetes mellitus

Abstract

Coptis chinensis Franch. and Panax ginseng C. A. Mey. are traditional herbal medicines with millennia of documented use and broad therapeutic applications, including anti-diabetic properties. However, the synergistic effect of total alkaloids from Coptis chinensis and total ginsenosides from Panax ginseng on type 2 diabetes mellitus (T2DM) and its underlying mechanism remain unclear. The research demonstrated that the optimal ratio of total alkaloids from Coptis chinensis and total ginsenosides from Panax ginseng was 4∶1, exhibiting maximal efficacy in improving insulin resistance and gluconeogenesis in primary mouse hepatocytes. This combination demonstrated significant synergistic effects in improving glucose tolerance, reducing fasting blood glucose (FBG), the weight ratio of epididymal white adipose tissue (eWAT), and the homeostasis model assessment of insulin resistance (HOMA-IR) in leptin receptor-deficient (db/db) mice. Subsequently, a T2DM liver-specific network was constructed based on RNA sequencing (RNA-seq) experiments and public databases by integrating transcriptional properties of disease-associated proteins and protein-protein interactions (PPIs). The network recovery index (NRI) score of the combined treatment group with a 4∶1 ratio exceeded that of groups treated with individual components. The research identified that activated adenosine 5'-monophosphate-activated protein kinase (AMPK)/acetyl-CoA carboxylase (ACC) signaling in the liver played a crucial role in the synergistic treatment of T2DM, as verified by western blot experiment in db/db mice. These findings demonstrate that the 4∶1 combination of total alkaloids from Coptis chinensis and total ginsenosides from Panax ginseng significantly improves insulin resistance and glucose and lipid metabolism disorders in db/db mice, surpassing the efficacy of individual treatments. The synergistic mechanism correlates with enhanced AMPK/ACC signaling pathway activity.

Keywords

Total alkaloids from Coptis chinensis / Alkaloids / Total ginsenosides from Panax ginseng / Component compatibility / Network pharmacology / Type 2 diabetes mellitus

References

[1] Ogurtsova K, Guariguata L, Barengo NC, et al. IDF Diabetes Atlas: global estimates of undiagnosed diabetes in adults for 2021. Diabetes Res Clin Pract. 2022;183:109118. https://doi.org/10.1016/j.diabres.2021.109118.

[2] Dendup T, Feng X, Clingan S, et al. Environmental risk factors for developing type 2 diabetes mellitus: a systematic review. Int J Environ Res Public Health. 2018; 15(1):78. https://doi.org/10.3390/ijerph15010078.

[3] Wang L, Peng W, Zhao Z, et al.Prevalence and treatment of diabetes in China, 2013-2018. JAMA. 2021; 326(24):2498-2506. https://doi.org/10.1001/jama.2021.22208.

[4] Chan JCN, Lim LL, Wareham NJ, et al. The Lancet Commission on Diabetes: using data to transform diabetes care and patient lives. Lancet. 2021; 396(10267):2019-2082. https://doi.org/10.1016/S0140-6736(20)32374-6.

[5] Saeedi P, Salpea P, Karuranga S, et al. Mortality attributable to diabetes in 20-79 years old adults, 2019 estimates: results from the International Diabetes Federation Diabetes Atlas, 9^th edition. Diabetes Res Clin Pract. 2020;162:108086. https://doi.org/10.1016/j.diabres.2020.108086.

[6] Bianchi C, Daniele G, Dardano A, et al. Early combination therapy with oral glucose-lowering agents in type 2 diabetes. Drugs. 2017; 77(3):247-264. https://doi.org/10.1007/s40265-017-0694-4.

[7] Del Prato S. Rational combination therapy for type 2 diabetes. Lancet Diabetes Endocrinol. 2019; 7(5):328-329. https://doi.org/10.1016/S2213-8587(19)30069-5.

[8] Hu RF, Sun XB. Design of new traditional Chinese medicine herbal formulae for treatment of type 2 diabetes mellitus based on network pharmacology. Chin J Nat Med. 2017; 15(6):436-441. https://doi.org/10.1016/s1875-5364(17)30065-1.

[9] Wang H, Shi S, Wang S. Can highly cited herbs in ancient traditional Chinese medicine formulas and modern publications predict therapeutic targets for diabetes mellitus? J Ethnopharmacol. 2018; 213:101-110. https://doi.org/10.1016/j.jep.2017.10.032.

[10] Li JC, Shen XF, Meng XL. A traditional Chinese medicine JiuHuangLian (Rhizoma Coptidis steamed with rice wine) reduces oxidative stress injury in type 2 diabetic rats. Food Chem Toxicol. 2013; 59:222-229. https://doi.org/10.1016/j.fct.2013.06.005.

[11] Li JC, Shen XF, Shao JA, et al. The total alkaloids from Coptis chinensis Franch improve cognitive deficits in type 2 diabetic rats. Drug Des Devel Ther. 2018; 12:2695-2706. https://doi.org/10.2147/DDDT.S171025.

[12] Zhang H, Wei J, Xue R, et al. Berberine lowers blood glucose in type 2 diabetes mellitus patients through increasing insulin receptor expression. Metabolism. 2010; 59(2):285-292. https://doi.org/10.1016/j.metabol.2009.07.029.

[13] Zhang Y, Gu Y, Ren H, et al. Gut microbiome-related effects of berberine and probiotics on type 2 diabetes (the PREMOTE study). Nat Commun. 2020; 11(1):5015. https://doi.org/10.1038/s41467-020-18414-8.

[14] Sun Y, Xia M, Yan H, et al.Berberine attenuates hepatic steatosis and enhances energy expenditure in mice by inducing autophagy and fibroblast growth factor 21. Br J Pharmacol. 2018; 175(2):374-387. https://doi.org/10.1111/bph.14079.

[15] Zhao MM, Lu J, Li S, et al. Berberine is an insulin secretagogue targeting the KCNH6 potassium channel. Nat Commun. 2021; 12(1):5616. https://doi.org/10.1038/s41467-021-25952-2.

[16] Liu C, Zhang M, Hu MY, et al. Increased glucagon-like peptide-1 secretion may be involved in antidiabetic effects of ginsenosides. J Endocrinol. 2013; 217(2):185-196. https://doi.org/10.1530/JOE-12-0502.

[17] Liu Q, Zhang FG, Zhang WS, et al. Ginsenoside Rg1 inhibits glucagon-induced hepatic gluconeogenesis through Akt-FoxO1 interaction. Theranostics. 2017; 7(16):4001-4012. https://doi.org/10.7150/thno.18788.

[18] Zhou R, He D, Zhang H, et al. Ginsenoside Rb 1 protects against diabetes-associated metabolic disorders in Kkay mice by reshaping gut microbiota and fecal metabolic profiles. J Ethnopharmacol. 2023;303:115997. https://doi.org/10.1016/j.jep.2022.115997.

[19] Demir S, Nawroth PP, Herzig S, et al. Emerging targets in type 2 diabetes and diabetic complications. Adv Sci (Weinh). 2021; 8(18):e2100275. https://doi.org/10.1002/advs.202100275.

[20] Ma LL, Yuan YY, Zhao M, et al. Mori Cortex extract ameliorates nonalcoholic fatty liver disease (NAFLD) and insulin resistance in high-fat-diet/streptozotocin-induced type 2 diabetes in rats. Chin J Nat Med. 2018; 16(6):411-417. https://doi.org/10.1016/S1875-5364(18)30074-8.

[21] Wojcikowski K, Gobe G. Animal studies on medicinal herbs: predictability, dose conversion and potential value. Phytother Res. 2014; 28(1):22-27. https://doi.org/10.1002/ptr.4966.

[22] Shen W, Chuang CC, Martinez K, et al. Conjugated linoleic acid reduces adiposity and increases markers of browning and inflammation in white adipose tissue of mice. J Lipid Res. 2013; 54(4):909-922. https://doi.org/10.1194/jlr.M030924.

[23] Liao J, Hao C, Huang W, et al. Network pharmacology study reveals energy metabolism and apoptosis pathways-mediated cardioprotective effects of Shenqi Fuzheng. J Ethnopharmacol. 2018; 227:155-165. https://doi.org/10.1016/j.jep.2018.08.029.

[24] Fang S, Dong L, Liu L, et al. HERB: a high-throughput experiment- and reference-guided database of traditional Chinese medicine. Nucleic Acids Res. 2021; 49(D1):D1197-D1206. https://doi.org/10.1093/nar/gkaa1063.

[25] Kong X, Liu C, Zhang Z, et al. BATMAN-TCM 2.0:an enhanced integrative database for known and predicted interactions between traditional Chinese medicine ingredients and target proteins. Nucleic Acids Res. 2024; 52(D1):D1110-D1120. https://doi.org/10.1093/nar/gkad926.

[26] Ngo LP, Owiti NA, Swartz C, et al. Sensitive CometChip assay for screening potentially carcinogenic DNA adducts by trapping DNA repair intermediates. Nucleic Acids Res. 2020; 48(3):e13. https://doi.org/10.1093/nar/gkz1077.

[27] Yan J, Wang C, Jin Y, et al. Catalpol ameliorates hepatic insulin resistance in type 2 diabetes through acting on AMPK/NOX4/PI3K/AKT pathway. Pharmacol Res. 2018; 130:466-480. https://doi.org/10.1016/j.phrs.2017.12.026.

[28] Liu L, Wang ZB, Song Y, et al.Simultaneous determination of eight alkaloids in rat plasma by UHPLC-MS/MS after oral administration of Coptis deltoidea C. Y. Cheng et Hsiao and Coptis chinensis Franch. Molecules. 2016; 21(7):913. https://doi.org/10.3390/molecules21070913.

[29] Feng X, Wang K, Hu X, et al. Systematic screening and characterization of absorbed constituents and in vivo metabolites in rats after oral administration of Rhizoma Coptidis using UPLC-Q-TOF/MS. Biomed Chromatogr. 2020; 34(10):e4919. https://doi.org/10.1002/bmc.4919.

[30] Liu Y, Zhang Y, Meng Q, et al. Metabolic profile of alkaloids in Rhizoma Coptidis in rat plasma, urine and feces after oral administration using ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. Rapid Commun Mass Spectrom. 2020; 34(9):e8763. https://doi.org/10.1002/rcm.8763.

[31] Feng X, Wang K, Cao S, et al. Pharmacokinetics of five alkaloids and their metabolites in normal and diabetic rats after oral administration of Rhizoma Coptidis. Planta Med. 2022; 88(11):921-932. https://doi.org/10.1055/a-1506-1627.

[32] Dong WW, Han XZ, Zhao J, et al. Metabolite profiling of ginsenosides in rat plasma, urine and feces by LC-MS/MS and its application to a pharmacokinetic study after oral administration of Panax ginseng extract. Biomed Chromatogr. 2018; 32(3):e4105. https://doi.org/10.1002/bmc.4105.

[33] Liu C, Yang T, Zhao Z, et al. Effects of particle size reduction combined with β-cyclodextrin on the in vitro dissolution and in vivo relative bioavailability of ginsenosides in Panax ginseng. Food Funct. 2022; 13(21):10882-10894. https://doi.org/10.1039/D2FO01098D.

[34] Chen Q, Xiao S, Li Z, et al. Chemical and metabolic profiling of Si-Ni Decoction analogous formulae by high performance liquid chromatography-mass spectrometry. Sci Rep. 2015;5:11638. https://doi.org/10.1038/srep11638.

[35] Miao WJ, Hu Y, Jia L, et al. Profiling and identification of chemical components of Shenshao Tablet and its absorbed components in rats by comprehensive HPLC/DAD/ESI-MSⁿ analysis. Chin J Nat Med. 2018; 16(10):791-800. https://doi.org/10.1016/S1875-5364(18)30119-5.

[36] Perng W, Conway R, Mayer-Davis E, et al. Youth-onset type 2 diabetes: the epidemiology of an awakening epidemic. Diabetes Care. 2023; 46(3):490-499. https://doi.org/10.2337/dci22-0046.

[37] Ma H, He K, Zhu J, et al. The anti-hyperglycemia effects of Rhizoma Coptidis alkaloids: a systematic review of modern pharmacological studies of the traditional herbal medicine. Fitoterapia. 2019; 134:210-220. https://doi.org/10.1016/j.fitote.2019.03.003.

[38] Abdelazim A, Khater S, Ali H, et al. Panax ginseng improves glucose metabolism in streptozotocin-induced diabetic rats through 5' adenosine monophosphate kinase up-regulation. Saudi J Biol Sci. 2019; 26(7):1436-1441. https://doi.org/10.1016/j.sjbs.2018.06.001.

[39] Li X, Liu Z, Liao J, et al. Network pharmacology approaches for research of traditional Chinese medicines. Chin J Nat Med. 2023; 21(5):323-332. https://doi.org/10.1016/S1875-5364(23)60429-7.

[40] Li M, Chi X, Wang Y, et al. Trends in insulin resistance: insights into mechanisms and therapeutic strategy. Signal Transduct Target Ther. 2022; 7(1):216. https://doi.org/10.1038/s41392-022-01073-0.

[41] Petersen MC, Vatner DF, Shulman GI. Regulation of hepatic glucose metabolism in health and disease. Nat Rev Endocrinol. 2017; 13(10):572-587. https://doi.org/10.1038/nrendo.2017.80.

[42] Rines AK, Sharabi K, Tavares CDJ, et al. Targeting hepatic glucose metabolism in the treatment of type 2 diabetes. Nat Rev Drug Discov. 2016; 15(11):786-804. https://doi.org/10.1038/nrd.2016.151.

[43] Perry RJ, Samuel VT, Petersen KF, et al. The role of hepatic lipids in hepatic insulin resistance and type 2 diabetes. Nature. 2014; 510(7503):84-91. https://doi.org/10.1038/nature13478.

[44] Martinez B, Ortiz RM. Thyroid hormone regulation and insulin resistance: insights from animals naturally adapted to fasting. Physiology (Bethesda). 2017; 32(2):141-151. https://doi.org/10.1152/physiol.00018.2016.

[45] Gauthier BR, Sola-García A, Cáliz-Molina M, et al. Thyroid hormones in diabetes, cancer, and aging. Aging Cell. 2020; 19(11):e13260. https://doi.org/10.1111/acel.13260.

[46] He F, Huang Y, Song Z, et al. Mitophagy-mediated adipose inflammation contributes to type 2 diabetes with hepatic insulin resistance. J Exp Med. 2021; 218(3):e20201416. https://doi.org/10.1084/jem.20201416.

[47] Mahboobifard F, Pourgholami MH, Jorjani M, et al. Estrogen as a key regulator of energy homeostasis and metabolic health. Biomed Pharmacother. 2022;156:113808. https://doi.org/10.1016/j.biopha.2022.113808.

[48] Yan H, Yang W, Zhou F, et al.Estrogen improves insulin sensitivity and suppresses gluconeogenesis via the transcription factor Foxo1. Diabetes. 2019; 68(2):291-304. https://doi.org/10.2337/db18-0638.

[49] De Paoli M, Zakharia A, Werstuck GH. The role of estrogen in insulin resistance: a review of clinical and preclinical data. Am J Pathol. 2021; 191(9):1490-1498. https://doi.org/10.1016/j.ajpath.2021.05.011.

[50] Joshi MB, Ahamed R, Hegde M, et al. Glucose induces metabolic reprogramming in neutrophils during type 2 diabetes to form constitutive extracellular traps and decreased responsiveness to lipopolysaccharides. Biochim Biophys Acta Mol Basis Dis. 2020; 1866(12):165940. https://doi.org/10.1016/j.bbadis.2020.165940.

[51] Yang S, Wang S, Chen L, et al. Neutrophil extracellular traps delay diabetic wound healing by inducing endothelial-to-mesenchymal transition via the Hippo pathway. Int J Biol Sci. 2023; 19(1):347-361. https://doi.org/10.7150/ijbs.78046.

[52] Lawan A, Bennett AM. Mitogen-activated protein kinase regulation in hepatic metabolism. Trends Endocrinol Metab. 2017; 28(12):868-878. https://doi.org/10.1016/j.tem.2017.10.007.

[53] Steinberg GR, Hardie DG. New insights into activation and function of the AMPK. Nat Rev Mol Cell Biol. 2023; 24(4):255-272. https://doi.org/10.1038/s41580-022-00547-x.

[54] Steneberg P, Lindahl E, Dahl U, et al. PAN-AMPK activator O304 improves glucose homeostasis and microvascular perfusion in mice and type 2 diabetes patients. JCI Insight. 2018; 3(12):e99114. https://doi.org/10.1172/jci.insight.99114.