Ipomoea Parasitica AERIAL-PART SOXHLET FRACTIONS: ANTIDIABETIC EFFICACY IN STREPTOZOTOCIN-INDUCED RATS AND REVERSAL OF PALMITATE-DRIVEN INSULIN RESISTANCE IN L6 MYOTUBES AND HEPG2 HEPATOCYTES
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Abstract
Standardized Soxhlet fractions of Ipomoea parasitica (Convolvulaceae) aerial parts hexane (HE), ethyl acetate (EAE),
ethanol (EE), and hydroalcoholic (HAE) were evaluated for acute oral toxicity and antidiabetic activity. Acute toxicity
screening by OECD 423 Acute Toxic Class method returned no mortality at 2000 mg/kg p.o. for EAE, EE, and HAE in
Swiss albino mice, establishing LD50 > 2000 mg/kg (GHS Category 5). A 28-day in vivo study in streptozotocin-induced
(55 mg/kg i.p.) Wistar rats (n = 6/group) dosed all three fractions at 400 mg/kg p.o. daily against metformin 150 mg/kg
p.o. as comparator. Day-28 fasting blood glucose reductions versus the diabetic control: EE −57.7% (p < 0.001), EAE
−45.5% (p < 0.001), HAE −35.5% (p < 0.01). EE and metformin (−61.2%) were not statistically separable by Tukey
HSD at n = 6. Body weight tracked the glucose response: EE gained +9.9% against a diabetic-control loss of −16.4%.
In vitro, MTT cytotoxicity established ≤100 µg/mL as the safe working ceiling (≥80% viability) for EE and EAE in both
L6 myotubes and HepG2 hepatocytes, and ≤50 µg/mL for HAE. Palmitate-induced insulin-resistant L6 myotubes (300
µM, 16 h) recovered 2-NBDG uptake to 167 ± 7% of unchallenged basal with EE 100 µg/mL (versus metformin 1 mM:
178 ± 6%; insulin-resistant baseline: 96 ± 5%). In HepG2 hepatocytes (250 µM palmitate, 24 h), EE 100 µg/mL reached
152 ± 7% versus metformin 158 ± 6%. EE EC50 values were 22 µg/mL in L6 and 31 µg/mL in HepG2. The cellular
reversal ranking EE > EAE > HAE mirrored the polyphenol-load hierarchy established in the companion phytochemistry
study. These findings constitute the first paired in vivo–in vitro antidiabetic record for I. parasitica at the aerial-part
level and position EE as a lead fraction warranting mechanistic extension.
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References
Aden, D. P., Fogel, A., Plotkin, S., Damjanov, I., & Knowles, B. B. (1979). Controlled synthesis of HBsAg in a differentiated human liver carcinoma-derived cell line. Nature, 282(5739), 615–616.
https://doi.org/10.1038/282615a0
Bailey, C. J., & Turner, R. C. (1996). Metformin. New England Journal of Medicine, 334(9), 574–579.
https://doi.org/10.1056/NEJM199602293340906
Chavez, J. A., & Summers, S. A. (2003). Characterizing the effects of saturated fatty acids on insulin signaling and ceramide and diacylglycerol accumulation in 3T3-L1 adipocytes and C2C12 myotubes. Archives of Biochemistry and Biophysics, 419(2), 101–109.
https://doi.org/10.1016/j.abb.2003.08.020
CPCSEA. (2003). CPCSEA guidelines for laboratory animal facility. Indian Journal of Pharmacology, 35(4), 257–274.
Dhanya, R., Arun, K. B., Syama, H. P., & Nisha, P. (2017). Rutin and quercetin enhance glucose uptake in L6 myotubes. Frontiers in Pharmacology, 8, 336.
https://doi.org/10.3389/fphar.2017.00336
Dunnett, C. W. (1955). A multiple comparison procedure for comparing several treatments with a control. Journal of the American Statistical Association, 50(272), 1096–1121.
Eid, H. M., & Haddad, P. S. (2017). The antidiabetic mechanisms of quercetin. Current Medicinal Chemistry, 24(4), 355–364.
https://doi.org/10.2174/0929867323666160909153707
Furman, B. L. (2021). Streptozotocin-induced diabetic models in mice and rats. Current Protocols, 1(4), e78. https://doi.org/10.1002/cpz1.78
Goyal, S. N., Reddy, N. M., Patil, K. R., et al. (2016). Challenges and issues with streptozotocin-induced diabetes. Chemico-Biological Interactions, 244, 49–63.
https://doi.org/10.1016/j.cbi.2015.11.032
Hardie, D. G., Schaffer, B. E., & Brunet, A. (2016). AMPK: An energy-sensing pathway with multiple inputs and outputs. Trends in Cell Biology, 26(3), 190–201.
https://doi.org/10.1016/j.tcb.2015.10.013
Hirosumi, J., Tuncman, G., Chang, L., et al. (2002). A central role for JNK in obesity and insulin resistance. Nature, 420(6913), 333–336.
https://doi.org/10.1038/nature01137
Hsu, C. Y., Shih, H. Y., Chia, Y. C., et al. (2014). Rutin potentiates insulin receptor kinase to enhance insulin-dependent glucose transport activity. Molecular Nutrition & Food Research, 58(6), 1168–1176. https://doi.org/10.1002/mnfr.201300691
Huang, D. W., Chang, W. C., Wu, J. S. B., Shih, R. W., & Shen, S. C. (2016). Gallic acid ameliorates hyperglycemia and improves hepatic carbohydrate metabolism in rats fed a high-fructose diet. Nutrition Research, 36(2), 150–160. https://doi.org/10.1016/j.nutres.2015.10.001
Ishii, Y., Maeda, A., Tani, K., & Akagawa, M. (2015). Palmitate induces insulin resistance in human HepG2 hepatocytes by enhancing ubiquitination and proteasomal degradation of key insulin signaling molecules. Archives of Biochemistry and Biophysics, 566, 26–35.
https://doi.org/10.1016/j.abb.2014.12.009
Jiang, S., Young, J. L., Wang, K., Qian, Y., & Ai, L. (2019). Quercetin-induced AMPK activation: A potential treatment strategy for type 2 diabetes. Scientific Reports, 9, 38989.
https://doi.org/10.1038/s41598-019-38989-7
Kamalakkannan, N., & Prince, P. S. M. (2006). Antihyperglycaemic and antioxidant effect of rutin in streptozotocin-induced diabetic Wistar rats. Basic & Clinical Pharmacology & Toxicology, 98(1), 97–103.
https://doi.org/10.1111/j.17427843.2006.pto
Kjøbsted, R., Hingst, J. R., Fentz, J., et al. (2018). AMPK in skeletal muscle function and metabolism. FASEB Journal, 32(4), 1741–1777.
https://doi.org/10.1096/fj.201700442R
Knowles, B. B., Howe, C. C., & Aden, D. P. (1980). Human hepatocellular carcinoma cell lines secrete the major plasma proteins and hepatitis B surface antigen. Science, 209(4455), 497–499.
https://doi.org/10.1126/science.6248960
Lenzen, S. (2008). The mechanisms of alloxan- and streptozotocin-induced diabetes. Diabetologia, 51(2), 216–226. https://doi.org/10.1007/s00125-007-0886-7
Listenberger, L. L., Han, X., Lewis, S. E., et al. (2003). Triglyceride accumulation protects against fatty acid-induced lipotoxicity. PNAS, 100(6), 3077–3082. https://doi.org/10.1073/pnas.0630588100
Malalavidhane, T. S., Wickramasinghe, S. M., & Jansz, E. R. (2000). Oral hypoglycaemic activity of Ipomoea aquatica. Journal of Ethnopharmacology, 72(1–2), 293–298.
https://doi.org/10.1016/S0378-8741(00)00207-X
Malalavidhane, T. S., Wickramasinghe, S. M., Perera, M. S., & Jansz, E. R. (2003). Oral hypoglycaemic activity of Ipomoea aquatica in streptozotocin-induced, diabetic Wistar rats and type II diabetics. Phytotherapy Research, 17(9), 1098–1100.
https://doi.org/10.1002/ptr.1345
Mitsumoto, Y., & Klip, A. (1992). Development regulation of GLUT1 and GLUT4 glucose transporters during myogenesis of L6 muscle cells. Journal of Biological Chemistry, 267(7), 4957–4962.
Mosmann, T. (1983). Rapid colorimetric assay for cellular growth and survival. Journal of Immunological Methods, 65(1–2), 55–63.
https://doi.org/10.1016/0022-1759(83)90303-4
Nakamura, S., Takamura, T., Matsuzawa-Nagata, N., et al. (2009). Palmitate induces insulin resistance in H4IIEC3 hepatocytes through reactive oxygen species produced by mitochondria. Journal of Biological Chemistry, 284(22), 14809–14818.
https://doi.org/10.1074/jbc.M901488200
OECD. (2002). Test No. 423: Acute oral toxicity Acute toxic class method. OECD Publishing.
https://doi.org/10.1787/9789264071001-en
Percie du Sert, N., Hurst, V., Ahluwalia, A., et al. (2020). The ARRIVE guidelines 2.0. PLOS Biology, 18(7), e3000410.
https://doi.org/10.1371/journal.pbio.3000410
Powell, D. J., Turban, S., Gray, A., Hajduch, E., & Hundal, H. S. (2004). Intracellular ceramide synthesis and protein kinase Cζ activation play an essential role in palmitate-induced insulin resistance in rat L6 skeletal muscle cells. Biochemical Journal, 382, 619–629.
https://doi.org/10.1042/BJ20040139
Reaven, G. M. (1988). Role of insulin resistance in human disease. Diabetes, 37(12), 1595–1607. https://doi.org/10.2337/diab.37.12.1595
Rena, G., Hardie, D. G., & Pearson, E. R. (2017). The mechanisms of action of metformin. Diabetologia, 60(9), 1577–1585.
https://doi.org/10.1007/s00125-017-4342-z
Sánchez-Rangel, J. C., Benavides, J., Heredia, J. B., Cisneros-Zevallos, L., & Jacobo-Velázquez, D. A. (2013). The Folin–Ciocalteu assay revisited. Analytical Methods, 5(21), 5990–5999.
https://doi.org/10.1039/C3AY41125G
Sajak, A. A. B., Abas, F., & Ismail, A. (2017). Ipomoea aquatica ameliorates glucose metabolism. Phytomedicine, 33, 1–8. https://doi.org/10.1016/j.phymed.2017.10.011
Tukey, J. W. (1949). Comparing individual means in the analysis of variance. Biometrics, 5(2), 99–114.
Vijayakumar, S., Presannakumar, G., & Vijayalakshmi, N. R. (2012). Antidiabetic activity of Ipomoea aquatica in alloxan-induced diabetic rats. Journal of Ethnopharmacology, 139(2), 396–400.
https://doi.org/10.1016/j.jep.2012.02.005
Wilcox, G. (2005). Insulin and insulin resistance. Clinical Biochemist Reviews, 26(2), 19–39.
Xiao, J., & Högger, P. (2015). Dietary polyphenols and type 2 diabetes. Current Medicinal Chemistry, 22(1), 23–38.
https://doi.org/10.2174/0929867321666140706130807
Yaffe, D. (1968). Retention of differentiation potentialities during prolonged cultivation of myogenic cells. PNAS, 61(2), 477–483.
https://doi.org/10.1073/pnas.61.2.477
Yamada, K., Saito, M., Matsuoka, H., & Inagaki, N. (2007). A real-time method of imaging glucose uptake in single, living mammalian cells. Nature Protocols, 2(3), 753–762.
https://doi.org/10.1038/nprot.2007.76
Yuan, M., Konstantopoulos, N., Lee, J., et al. (2001). Reversal of obesity- and diet-induced insulin resistance with salicylates or targeted disruption of Ikkβ. Science, 293(5535), 1673–1677.