R-type ginsenoside Rg3 attenuates cisplatin-induce intestinal injury via restoring autophagy flux blockade

Abstract

Cisplatin and its metabolites can cause severe gastrointestinal mucosal damage, leading to varying degrees of intestinal injury in nearly all patients following administration. Although the anti-tumor effectiveness of R-ginsenoside Rg3 (R-Rg3, a key chemical component of Shen Yi Capsule) as the first-line drug is widely recognized in the clinic, there is less concern about the improved effects of R-Rg3 against intestinal toxicity caused by concurrent cisplatin chemotherapy. This study aimed to investigate the protective effects of R-Rg3 against cisplatin-induced intestinal toxicity and to explore its potential molecular targets and mechanisms. Rats in the R-Rg3 treatment group were treated with R-Rg3 (7, 14 mg·kg⁻¹, p.o.) for 7 days, and a single dose of cisplatin (14 mg·kg⁻¹, i.p.) was administered via intraperitoneal injection to rats in the cisplatin group and R-Rg3 treatment group on the 7^th day. Intestinal epithelial cell line 6 (IEC-6) were pretreated with R-Rg3 (1.25, 2.5, 5 μmol·L⁻¹) for 24 h followed by cisplatin treatment (3 μmol·L⁻¹, 24 h). The in vivo results showed that, R-Rg3 treatment for 7 days markedly alleviated cisplatin-induced oxidative stress and mitochondrial dysfunction in vivo, while suppressing excessive autophagy and reducing intestinal damage. In IEC-6 cells, R-Rg3 pretreatment inhibited mitophagy and promoted the restoration of lysosomal function. Autophagy inhibitors 3-methyladenine (autophagosome formation inhibitor, 1 μmol·L⁻¹) and Bafilomycin A1 (proton pump inhibitor, 8 μmol·L⁻¹) were used to verify the mechanism of R-Rg3 action. Importantly, application of 3-methyladenine and Bafilomycin A1 verified that R-Rg3 could alleviate gastrointestinal dysfunction by restoring the cisplatin-induced autophagic flux blockade. In conclusion, this study identifies a previously underappreciated protective role of R-Rg3 against cisplatin-induced intestinal injury. Furthermore, it suggests that pharmacological modulation of the mitochondria-lysosome axis by R-Rg3 may effectively mitigate oxidative stress-mediated autophagic flux impairment caused by cisplatin.

Keywords

R-Ginsenoside Rg3 / Cisplatin / Lysosomal dysfunction / Autophagic flux blockade.

Funding

This work was supported by the National NaturalScience Foundation of China (No. 82474070), the National Key Research and Development Program (No. 2023YFD1601600) and the Science and Technology Research Project of Jilin Provincial Department of Education (No. JJKH20210375KJ).

Declaration of competing interest

The authors declare that there are no conflicts of interest.

References

[1] Dasari S, Tchounwou PB. Cisplatin in cancer therapy: molecularmechanisms of action. Eur J Pharmacol. 2014; 740:364-378. https://doi.org/10.1016/j.ejphar.2014.07.025.

[2] Qi LY, Luo Q, Zhang YY, et al. Advances in toxicological research of the anticancer drug cisplatin. Chem Res Toxicol. 2019; 32(8):1469-1486. https://doi.org/10.1021/acs.chemrestox.9b00204.

[3] Soodvilai S, Tipparos W, Rangsimawong W, et al. Effects of silymarin-loaded amphiphilic chitosan polymeric micelles on the renal toxicity and anticanceractivity of cisplatin. Pharm Dev Technol. 2019; 24(8):927-934. https://doi.org/10.1080/10837450.2018.1556690.

[4] Ghosh S. Cisplatin: the first metal based anticancer drug. Bioorganic Chem. 2019; 88:102925. https://doi.org/10.1016/j.bioorg.2019.102925.

[5] Shahid F, Farooqui Z, Khan F. Cisplatin-induced gastrointestinal toxicity: an update on possible mechanisms and on available gastroprotectivestrategies. Eur J Pharmacol. 2018; 827:49-57. https://doi.org/10.1016/j.ejphar.2018.03.009.

[6] Joyce K, Saxena S, Williams A, et al. Antimicrobial spectrum of the antitumor agent, cisplatin. J Antibiot. 2010; 63(8):530-532. https://doi.org/10.1038/ja.2010.64.

[7] Shim HS, Bae C, Wang J, et al. Peripheral and central oxidative stress in chemotherapy-induced neuropathic pain. Mol Pain. 2019; 15:1744806919840098. https://doi.org/10.1177/1744806919840098.

[8] Wang Z, Dong Y, Bai YY, et al. Platycodin D improves early atherosclerosis in type 2 diabetes mellitus by regulating endothelial inflammation and apoptosis. Am J Chin Med. 2025:1-26. https://doi.org/10.1142/S0192415X25500703.

[9] Suzuki M, Bandoski C, Bartlett JD. Fluoride induces oxidative damage and SIRT1/autophagy through ROS-mediated JNK signaling. Free Radic BiolMed. 2015; 89:369-378. https://doi.org/10.1016/j.freeradbiomed.2015.08.015.

[10] Zhang JJ, Wang JQ, Xu XY, et al. Red ginseng protects against cisplatin-induced intestinal toxicity by inhibiting apoptosis and autophagy via the PI3K/AKT and MAPK signaling pathways. Food Funct. 2020; 11(5):4236-4248. https://doi.org/10.1039/d0fo00469c.

[11] Cheng XY, Liang JH, Wu D, et al. Blunting ROS/TRPML1 pathway protects AFB1-induced porcine intestinal epithelial cells apoptosis by restoring impaired autophagic flux. Ecotox Environ Safe. 2023; 257:114942. https://doi.org/10.1016/j.ecoenv.2023.114942.

[12] Mireault M, Xiao Y, Barbeau B, et al. Cadmium affects autophagy in the human intestinal cells Caco-2 through ROS-mediated ERK activation. Cell Biol Toxicol. 2023; 39(3):945-966. https://doi.org/10.1007/s10565-021-09655-4.

[13] Scherz Shouval R, Elazar Z. Regulation of autophagy by ROS: physiologyand pathology. Trends Biochem Sci. 2011; 36(1):30-38. https://doi.org/10.1016/j.tibs.2010.07.007.

[14] Ballabio A, Bonifacino JS. Lysosomes as dynamic regulators of cell and organismal homeostasis. Nat Rev Mol Cell Biol. 2020; 21(2):101-118. https://doi.org/10.1038/s41580-019-0185-4.

[15] Rashid S, Nafees S, Siddiqi A, et al. Partial protection by 18β-glycrrhetinic acid against cisplatin induced oxidative intestinal damage in wistar rats: possiblerole of NF-κB and caspases. Pharmacol Rep. 2017; 69(5):1007-1013. https://doi.org/10.1016/j.pharep.2017.02.013.

[16] Shi ZY, Zeng JZ, Wong AS. Chemical structures and pharmacological profiles of Ginseng saponins. Molecules. 2019; 24(13):2443. https://doi.org/10.3390/molecules24132443.

[17] Li K, Wang YJ, Chen C, et al. Targeting pyroptosis: a novel strategy of ginseng for the treatment of diabetes and its chronic complications. Phytomedicine. 2025; 138:156430. https://doi.org/10.1016/j.phymed.2025.156430.

[18] Shin JH, Park YJ, Kim W, et al. Change of ginsenoside profiles in processed ginseng by drying, steaming, and puffing. J Microbiol Biotechnol. 2019; 29(2):222-229. https://doi.org/10.4014/jmb.1809.09056.

[19] Peng M, Li XN, Zhang T, et al. Stereoselective pharmacokinetic and metabolism studies of 20(S)- and 20(R) -ginsenoside Rg₃epimers in rat plasma by liquid chromatography-electrospray ionizationmass spectrometry. J Pharm Biomed Anal. 2016; 121:215-224. https://doi.org/10.1016/j.jpba.2016.01.020.

[20] Li W, Wang JQ, Zhou YD, et al. Rare ginsenoside 20(R)-Rg3 inhibits D-galactose-induced liver and kidney injury by regulating oxidative stress-induced apoptosis. Am J Chin Med. 2020; 48(5):1141-1157. https://doi.org/10.1142/s0192415x20500561.

[21] Sha JY, Zhou YD, Yang JY, et al. Maltol (3-hydroxy-2-methyl-4-pyrone) slows D-galactose-induced brain aging process by damping the Nrf2/HO-1-mediated oxidative stress in mice. J Agric Food Chem. 2019; 67(37):10342-10351. https://doi.org/10.1021/acs.jafc.9b04614.

[22] Zhang JJ, Zhou YD, Liu YB, et al. Protective effect of 20(R)-ginsenoside Rg3 against cisplatin-induced renal toxicity via PI3K/AKT and NF-κB signaling pathways based on the premise of ensuring anticancer effect. Am J Chin Med. 2021; 49(07):1739-1756. https://doi.org/10.1142/s0192415x21500828.

[23] Hu JN, Yang JY, Jiang S, et al. Panax quinquefolium saponins protect against cisplatin evoked intestinal injury via ROS-mediated multiple mechanisms. Phytomedicine. 2021; 82:153446. https://doi.org/10.1016/j.phymed.2020.153446.

[24] Xing JJ, Hou JG, Ma ZN, et al. Ginsenoside Rb3 provides protective effects against cisplatin-induced nephrotoxicity via regulation of AMPK-/mTOR-mediated autophagy and inhibition of apoptosis in vitro and in vivo. Cell Prolif. 2019; 52(4):e12627. https://doi.org/10.1111/cpr.12627.

[25] Zhang JJ, Chen KC, Zhou Y, et al. Evaluating the effects of mitochondrial autophagy flux on ginsenoside Rg2 for delaying D-galactose induced brain aging in mice. Phytomedicine. 2022; 104:154341. https://doi.org/10.1016/j.phymed.2022.154341.

[26] Zhang YB, Sun H, Pei RJ, et al. The SARS-CoV-2 protein ORF3a inhibits fusion of autophagosomes with lysosomes. Cell Discov. 2021; 7(1):31. https://doi.org/10.1038/s41421-021-00268-z.

[27] Saad AA, Youssef MI, El-Shennawy LK. Cisplatin induced damage in kidney genomic DNA and nephrotoxicity in male rats: the protective effect of grape seed proanthocyanidin extract. Food Chem Toxicol. 2009; 47(7):1499-1506. https://doi.org/10.1016/j.fct.2009.03.043.

[28] Wu CH, Ko JL, Liao JM, et al. D-methionine alleviates cisplatin-induced mucositis by restoring the gut microbiota structure and improving intestinal inflammation. Ther Adv Med Oncol. 2019; 11:1758835918821021. https://doi.org/10.1177/1758835918821021.

[29] Del Re M, Latiano T, Fidilio L, et al. Unusual gastrointestinal and cutaneous toxicities by bleomycin, etoposide, and cisplatin: a case report with pharmacogenetic analysis to personalize treatment. Epma J. 2017; 8(1):69-73. https://doi.org/10.1007/s13167-017-0080-z.

[30] Chang BJ, Nishikawa M, Sato E, et al. L-Carnitine inhibits cisplatin-induced injury of the kidney and small intestine. Arch Biochem Biophys. 2002; 405(1):55-64. https://doi.org/10.1016/s0003-9861(02)00342-9.

[31] Li BW, Wang WZ, Li Z, et al. Micro RNA-148a-3p enhances cisplatin cytotoxicity in gastric cancer through mitochondrial fission induction and cyto-protective autophagy suppression. Cancer Lett. 2017; 410:212-227. https://doi.org/10.1016/j.canlet.2017.09.035.

[32] Guerrero-Beltran CE, Calderon-Oliver M, Martinez-Abundis E, et al. Protective effect of sulforaphane against cisplatin-induced mitochondrial alterations and impairment in the activity of NAD(P)H: quinone oxidoreductase 1 and γ glutamyl cysteine ligase: studies in mitochondria isolated from rat kidney and in LLC-PK1 cells. Toxicol Lett. 2010; 199(1):80-92. https://doi.org/10.1016/j.toxlet.2010.08.009.

[33] Xiao L, Cui T, Liu S, et al. Vitamin A supplementation improves the intestinal mucosal barrier and facilitates the expression of tight junction proteins inrats with diarrhea. Nutrition. 2019; 57:97-108. https://doi.org/10.1016/j.nut.2018.06.007.

[34] Obermeier B, Daneman R, Ransohoff RM. Development, maintenance and disruption of the blood-brain barrier. Nat Med. 2013; 19(12):1584-1596. https://doi.org/10.1038/nm.3407.

[35] Al-Sadi R, Khatib K, Guo S, et al. Occludin regulates macromolecule flux across the intestinal epithe lial tight junction barrier. Am J Physiol Gastroint Liver Physiol. 2011; 300(6):G1054-G1064. https://doi.org/10.1152/ajpgi.00055.2011.

[36] Hazman O, Bozkurt MF, Fidan AF, et al. The effect of boric acid and borax on oxidative stress, inflammation, ER stress and apoptosis in cisplatin toxication and nephrotoxicity developing as a result of toxication. Inflammation. 2018; 41(3):1032-1048. https://doi.org/10.1007/s10753-018-0756-0.

[37] Ma ZN, Li YZ, Li W, et al. Nephroprotective effects of saponins from leaves of Panax quinquefolius against cisplatin-induced acute kidney injury. Int J Mol Sci. 2017; 18(7):1407. https://doi.org/10.3390/ijms18071407.

[38] Ni YD, Zhu YY, Xu LX, et al. Pharmacological activities and mechanisms of proteins and peptides derived from traditional Chinese medicine. Sci Tradit Chin Med. 2024; 2(4):260-275. https://doi.org/10.1097/st9.0000000000000054.

[39] Deshmukh P, Unni S, Krishnappa G, et al. The Keap1-Nrf2 pathway: promising therapeutic target to counteract ROS-mediated damage in cancersand neurodegenerative diseases. Biophys Rev. 2017; 9(1):41-56. https://doi.org/10.1007/s12551-016-0244-4.

[40] Luo Q, Peng B, Wei XL, et al. Research progress of Chinese herbal medicines and classic prescriptions in the treatment of COPD based on theTCM theoryand medicinal characteristics. Sci Tradit Chin Med. 2024; 2(2):104-120. https://doi.org/10.1097/st9.0000000000000028.

[41] Lu YW, Xie LY, Qi MH, et al. Platycodin D ameliorates cognitive impairment in type 2 diabetes mellitus mice via regulating PI3K/Akt/GSK3β signalingpathway. J Agric Food Chem. 2024; 72(22):12516-12528. https://doi.org/10.1021/acs.jafc.3c08490.

[42] Yoshii SR, Mizushima N. Autophagy machinery in the context of mammalian mitophagy. Biochim Biophys Acta. 2015; 1853(10):2797-2801. https://doi.org/10.1016/j.bbamcr.2015.01.013.

[43] Mcwilliams TG, Muqit MMK. PINK1 and Parkin: emerging themes in mitochondrial homeostasis. Curr Opin Cell Biol. 2017; 45:83-91. https://doi.org/10.1016/j.ceb.2017.03.013.

[44] Galluzzi L, Baehrecke EH, Ballabio A, et al. Molecular definitions of autophagy and related processes. Embo J. 2017; 36(13):1811-1836. https://doi.org/10.15252/embj.201796697.

[45] Settembre C, Fraldi A, Medina DL, et al. Signals from the lysosome: a control centre for cellular clearance and energy metabolism. Nat Rev Mol Cell Biol. 2013; 14(5):283-296. https://doi.org/10.1038/nrm3565.

[46] Yu L, Chen Y, Tooze SA. Autophagy pathway: cellular and molecular mechanisms. Autophagy. 2018; 14(2):207-215. https://doi.org/10.1080/15548627.2017.1378838.