Abstract: Ganoderic triterpenoids (GTs) are the primary bioactive constituents of the Basidiomycotina fungus, Ganoderma lucidum. These compounds exhibit antitumor, anti-hyperlipidemic, and immune-modulatory pharmacological activities. This study focused on GT accumulation in mycelia of G. lucidum mediated by the heme oxygenase-1 (HO-1)/carbon monoxide (CO) signaling. Compared with the control, hemin (10 μmol/L) induced an increase of 60.1% in GT content and 57.1% in HO-1 activity. Moreover, carbon monoxide-releasing molecule-2 (CORM-2), CO donor, increased GT content by 56.0% and HO-1 activity by 18.1%. Zn protoporphyrin IX (ZnPPIX), a specific HO-1 inhibitor, significantly reduced GT content by 26.0% and HO-1 activity by 15.8%, while hemin supplementation reversed these effects. Transcriptome sequencing showed that HO-1/CO could function directly as a regulator involved in promoting GT accumulation by regulating gene expression in the mevalonate pathway, and modulating the reactive oxygen species (ROS) and Ca²⁺ pathways. The results of this study may help enhance large-scale GT production and support further exploration of GT metabolic networks and relevant signaling cross-talk.

血红素加氧酶-1/一氧化碳信号参与调控灵芝三萜积累的研究

[1]AhmadF, 2018. Ganoderma lucidum: persuasive biologically active constituents and their health endorsement. Biomed Pharmacother, 107:507-519.

[2]AmooaghaieR, TabatabaeiF, AhadiAM, 2015. Role of hematin and sodium nitroprusside in regulating Brassica nigra seed germination under nanosilver and silver nitrate stresses. Ecotoxicol Environ Saf, 113:259-270.

[3]BaiXG, ChenJH, KongXX, et al., 2012. Carbon monoxide enhances the chilling tolerance of recalcitrant Baccaurea ramiflora seeds via nitric oxide-mediated glutathione homeostasis. Free Radic Biol Med, 53(4):710-720.

[4]CaoZY, HuangBK, WangQY, et al., 2007. Involvement of carbon monoxide produced by heme oxygenase in ABA-induced stomatal closure in Vicia faba and its proposed signal transduction pathway. Chin Sci Bull, 52(17):2365-2373.

[5]CarotaG, RaffaeleM, SorrentiV, et al., 2019. Ginseng and heme oxygenase-1: the link between an old herb and a new protective system. Fitoterapia, 139:104370.

[6]ChenBZ, KeBR, YeLY, et al., 2017. Isolation and varietal characterization of Ganoderma resinaceum from areas of Ganoderma lucidum production in China. Sci Hortic, 224:109-114.

[7]CuiML, YangHY, HeGQ, 2015. Submerged fermentation production and characterization of intracellular triterpenoids from Ganoderma lucidum using HPLC-ESI-MS. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 16(12):998-1010.

[8]CuiML, YangHY, HeGQ, 2017. Apoptosis induction of colorectal cancer cells HTL-9 in vitro by the transformed products of soybean isoflavones by Ganoderma lucidum. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 18(12):1101-1112.

[9]DuanXL, DaiC, LiZW, et al., 2016. Ectopic over-expression of BoHO1, a cabbage heme oxygenase gene, improved salt tolerance in Arabidopsis: a case study on proteomic analysis. J Plant Physiol, 196-197:1-13.

[10]FatmawatiS, KondoR, ShimizuK, et al., 2013. Structure-activity relationships of lanostane-type triterpenoids from Ganoderma lingzhi as α‍-glucosidase inhibitors. Bioorg Med Chem Lett, 23(21):5900-5903.

[11]GuL, ZhongX, LianDH, et al., 2017. Triterpenoid biosynthesis and the transcriptional response elicited by nitric oxide in submerged fermenting Ganoderma lucidum. Process Biochem, 60:19-26.

[12]GuL, ZhengYM, LianDH, et al., 2018. Production of triterpenoids from Ganoderma lucidum: elicitation strategy and signal transduction. Process Biochem, 69:22-32.

[13]GuoK, KongWW, YangZM, 2009. Carbon monoxide promotes root hair development in tomato. Plant Cell Environ, 32(8):1033-1045.

[14]HanB, XuS, XieYJ, et al., 2012. ZmHO-1, a maize haem oxygenase-1 gene, plays a role in determining lateral root development. Plant Sci, 184:63-74.

[15]HanB, YangZ, XieYJ, et al., 2014. Arabidopsis HY1 confers cadmium tolerance by decreasing nitric oxide production and improving iron homeostasis. Mol Plant, 7(2):388-403.

[16]HanY, ZhangJ, ChenXY, et al., 2008. Carbon monoxide alleviates cadmium-induced oxidative damage by modulating glutathione metabolism in the roots of Medicago sativa. New Phytol, 177(1):155-166.

[17]HeHY, HeLF, 2014. The role of carbon monoxide signaling in the responses of plants to abiotic stresses. Nitric Oxide, 42:40-43.

[18]HuZP, DuRP, XiuL, et al., 2020. Protective effect of triterpenes of Ganoderma lucidum on lipopolysaccharide-induced inflammatory responses and acute liver injury. Cytokine, 127:154917.

[19]HuangJJ, HanB, XuS, et al., 2011. Heme oxygenase-1 is involved in the cytokinin-induced alleviation of senescence in detached wheat leaves during dark incubation. J Plant Physiol, 168(8):768-775.

[20]LianDH, LiL, LiuX, et al., 2021. Time-scale dynamics of proteome predicts the central carbon metabolism involved in triterpenoid accumulation responsive to nitrogen limitation in Ganoderma lucidum. Fungal Biol, 125(4):294-304.

[21]LinYT, LiMY, CuiWT, et al., 2012. Haem oxygenase-1 is involved in hydrogen sulfide-induced cucumber adventitious root formation. J Plant Growth Regul, 31(4):519-528.

[22]LiuH, WangZY, ZhouYC, et al., 2021. Immunomodulation of Chinese Herbal Medicines on NK cell populations for cancer therapy: a systematic review. J Ethnopharmacol, 268:113561.

[23]LiuYH, XuS, LingTF, et al., 2010. Heme oxygenase/carbon monoxide system participates in regulating wheat seed germination under osmotic stress involving the nitric oxide pathway. J Plant Physiol, 167(16):1371-1379.

[24]NieSP, ZhangH, LiWJ, et al., 2013. Current development of polysaccharides from Ganoderma: isolation, structure and bioactivities. Bioact Carbohydr Diet Fibre, 1(1):10-20.

[25]OtterbeinLE, BachFH, AlamJ, et al., 2000. Carbon monoxide has anti-inflammatory effects involving the mitogen-activated protein kinase pathway. Nat Med, 6(4):422-428.

[26]PengXR, LiL, DongJR, et al., 2019. Lanostane-type triterpenoids from the fruiting bodies of Ganoderma applanatum. Phytochemistry, 157:103-110.

[27]RenA, ShiL, ZhuJ, et al., 2019. Shedding light on the mechanisms underlying the environmental regulation of secondary metabolite ganoderic acid in Ganoderma lucidum using physiological and genetic methods. Fungal Genet Biol, 128:43-48.

[28]Santa-CruzD, PacienzaN, ZilliC, et al., 2017. Heme oxygenase up-regulation under ultraviolet-B radiation is not epigenetically restricted and involves specific stress-related transcriptions factors. Redox Biol, 12:549-557.

[29]SharmaC, BhardwajN, SharmaA, et al., 2019. Bioactive metabolites of Ganoderma lucidum: factors, mechanism and broad spectrum therapeutic potential. J Herb Med, 17-18:100268.

[30]ShiL, GongL, ZhangXY, et al., 2015. The regulation of methyl jasmonate on hyphal branching and GA biosynthesis in Ganoderma lucidum partly via ROS generated by NADPH oxidase. Fungal Genet Biol, 81:201-211.

[31]SinghN, BhatlaSC, 2016. Nitric oxide and iron modulate heme oxygenase activity as a long distance signaling response to salt stress in sunflower seedling cotyledons. Nitric Oxide, 53:54-64.

[32]SuHG, PengXR, ShiQQ, et al., 2020. Lanostane triterpenoids with anti-inflammatory activities from Ganoderma lucidum. Phytochemistry, 173:112256.

[33]TanXY, SunJS, NingHJ, et al., 2018. De novo transcriptome sequencing and comprehensive analysis of the heat stress response genes in the basidiomycetes fungus Ganoderma lucidum. Gene, 661:139-151.

[34]TianJL, RenA, WangT, et al., 2019. Hydrogen sulfide, a novel small molecule signalling agent, participates in the regulation of ganoderic acids biosynthesis induced by heat stress in Ganoderma lucidum. Fungal Genet Biol, 130:19-30.

[35]WangGH, XiaoY, DengXJ, et al., 2018. Exogenous hydrogen peroxide contributes to heme oxygenase-1 delaying programmed cell death in isolated aleurone layers of rice subjected to drought stress in a cGMP-dependent manner. Front Plant Sci, 9:84.

[36]WuMZ, WangFQ, ZhangC, et al., 2013. Heme oxygenase-1 is involved in nitric oxide- and cGMP-induced α-Amy2/54 gene expression in GA-treated wheat aleurone layers. Plant Mol Biol, 81(1-2):27-40.

[37]XieYJ, ZhangC, LaiDW, et al., 2014. Hydrogen sulfide delays GA-triggered programmed cell death in wheat aleurone layers by the modulation of glutathione homeostasis and heme oxygenase-1 expression. J Plant Physiol, 171(2):53-62.

[38]XuYN, ZhongJJ, 2012. Impacts of calcium signal transduction on the fermentation production of antitumor ganoderic acids by medicinal mushroom Ganoderma lucidum. Biotechnol Adv, 30(6):1301-1308.

[39]XuanW, XuS, LiMY, et al., 2012. Nitric oxide is involved in hemin-induced cucumber adventitious rooting process. J Plant Physiol, 169(11):1032-1039.

[40]ZhangG, RenA, ShiL, et al., 2018. Functional analysis of an APSES transcription factor (GlSwi6) involved in fungal growth, fruiting body development and ganoderic-acid biosynthesis in Ganoderma lucidum. Microbiol Res, 207:280-288.

[41]ZhangLP, ShaoHB, LongXH, et al., 2013. Gene regulation of iron-deficiency responses is associated with carbon monoxide and heme oxydase 1 in Chlamydomonas reinhardtii. PLoS ONE, 8(1):e53835.

[42]ZhangX, RenA, LiMJ, et al., 2016. Heat stress modulates mycelium growth, heat shock protein expression, ganoderic acid biosynthesis, and hyphal branching of Ganoderma lucidum via cytosolic Ca²⁺. Appl Environ Microbiol, 82(14):4112-4125.

[43]ZhaoXR, ZhangBJ, DengS, et al., 2016. Isolation and identification of oxygenated lanostane-type triterpenoids from the fungus Ganoderma lucidum. Phytochem Lett, 16:87-91.

[44]ZilliCG, Santa-CruzDM, BalestrasseKB, 2014. Heme oxygenase-independent endogenous production of carbon monoxide by soybean plants subjected to salt stress. Environ Exp Bot, 102:11-16.