Inonotus obliquus (Chaga) against HFD/STZ-induced glucolipid metabolism disorders and abnormal renal functions by regulating NOS-cGMP-PDE5 signaling pathway

Abstract

Our prior investigations have established that Inonotus obliquus (Chaga) possesses hypoglycemic effects. Persistent hyperglycemia is known to precipitate renal function abnormalities. The functionality of the kidneys is intricately linked to the levels of cyclic guanosine-3',5'-monophosphate (cGMP), which are influenced by the activities of nitric oxide synthase (NOS) and phosphodiesterase (PDE). Enhanced cGMP levels can be achieved either through the upregulation of NOS activity or the downregulation of PDE activity. The objective of the current study is to elucidate the effects of Chaga on disorders of glucolipid metabolism and renal abnormalities in rats with type 2 diabetes mellitus (T2DM), while concurrently examining the NOS-cGMP-PDE5 signaling pathway. A model of T2DM was developed in rats using a high-fat diet (HFD) combined with streptozotocin (STZ) administration, followed by treatment with Chaga extracts at doses of 50 and 100 mg·kg⁻¹ for eight weeks. The findings revealed that Chaga not only mitigated metabolic dysfunctions, evidenced by improvements in fasting blood glucose, total cholesterol, triglycerides, and insulin resistance, but also ameliorated renal function markers, including serum creatinine, urine creatinine (UCr), blood urea nitrogen, 24-h urinary protein, and estimated creatinine clearance. Additionally, enhancements in glomerular volume, GBM thickness, podocyte foot process width (FPW), and the mRNA and protein expressions of podocyte markers, such as nephrin and wilms tumor-1, were observed. Chaga was found to elevate cGMP levels in both serum and kidney tissues by increasing mRNA and protein expressions of renal endothelial NOS and neural NOS, while simultaneously reducing the expressions of renal inducible NOS and PDE5. In summary, Chaga counteracts HFD/STZ-induced glucolipid metabolism and renal function disturbances by modulating the NOS-cGMP-PDE5 signaling pathway. This research supports the potential application of Chaga in the clinical prevention and treatment of T2DM and diabetic nephropathy (DN), with cGMP serving as a potential therapeutic target.

Keywords

Inonotus obliquus / Diabetic nephropathy / Glucolipid metabolism disorders / Renal functions / NOS-cGMP-PDE5 signaling

References

[1] Kou RW, Han R, Gao YQ, et al. Anti-neuroinflammatory polyoxygenated lanostanoids from Chaga mushroomInonotus obliquus[J]. Phytochemistry, 2021, 184: 112647.

[2] Zhang Z, Liang X, Tong L, et al. Effect of Inonotus obliquus (Fr.) Pilat extract on the regulation of glycolipid metabolism via PI3K/Akt and AMPK/ACC pathways in mice[J]. J Ethnopharmacol, 2021, 273: 113963.

[3] Zhang Y, Liao H, Shen D, et al. Renal protective effects of Inonotus obliquus on high-fat diet/streptozotocin-induced diabetic kidney disease rats: biochemical, color doppler ultrasound and histopathological evidence[J]. Front Pharmacol, 2022, 12: 743931.

[4] Sohrabi M, Binaeizadeh MR, Iraji A, et al. A review on α-glucosidase inhibitory activity of first row transition metal complexes: a futuristic strategy for treatment of type 2 diabetes[J]. RSC Adv, 2022, 12(19): 12011-12052.

[5] Wong YH, Wong SH, Wong XT, et al. Genetic associated complications of type 2 diabetes mellitus[J]. Panminerva Med, 2022, 64(2): 274-288.

[6] Hua Q, Han Y, Zhao H, et al. Punicalagin alleviates renal injury via the gut-kidney axis in high-fat diet-induced diabetic mice[J]. Food Funct, 2022, 13(2): 867-879.

[7] Teh YM, Mualif SA, Lim SK. A comprehensive insight into autophagy and its potential signaling pathways as a therapeutic target in podocyte injury[J]. Int J Biochem Cell Biol, 2022, 143: 106153.

[8] Hu L, Chen Y, Zhou X, et al. Effects of soluble guanylate cyclase stimulator on renal function in ZSF-1 model of diabetic nephropathy[J]. PLoS One, 2022, 17(1): e0261000.

[9] Rogacka D, Rachubik P, Audzeyenka I, et al. Enhancement of cGMP-dependent pathway activity ameliorates hyperglycemia-induced decrease in SIRT1-AMPK activity in podocytes: impact on glucose uptake and podocyte function[J]. Biochim Biophys Acta Mol Cell Res, 2022, 1869(12): 119362.

[10] Ma L, Hu L, Jin L, et al. Rebalancing glucolipid metabolism and gut microbiome dysbiosis by nitrate-dependent alleviation of high-fat diet-induced obesity[J]. BMJ Open Diabetes Res Care, 2020, 8(1): e001255.

[11] Liao H, Jia D, Zhao X, et al. Effects of Chaga medicinal mushroom Inonotus obliquus (Agaricomycetes) extracts on NOS-cGMP-PDE5 pathway in rat penile smooth muscle cells [J]. Int J Med Mushrooms, 2020, 22(10): 979-990.

[12] Coskuner ER, Ozkan B. Reno-protective effects of phosphodiesterase 5 inhibitors[J]. Clin Exp Nephrol, 2021, 25(6): 585-597.

[13] Fanali C, Gallo V, Della PS, et al. Choline chloride-lactic acid-based NADES as an extraction medium in a response surface methodology-optimized method for the extraction of phenolic compounds from hazelnut skin[J]. Molecules, 2021, 26(9): 2652.

[14] Li W, Jiang L, Lu X, et al. Curcumin protects radiation-induced liver damage in rats through the NF-κB signaling pathway[J]. BMC Complement Med Ther, 2021, 21(1): 10.

[15] Zhang KQ, Tian T, Hu LL, et al. Effect of probucol on autophagy and apoptosis in the penile tissue of streptozotocin-induced diabetic rats[J]. Asian J Androl, 2020, 22(4): 409-413.

[16] Wilson RD, Islam MS. Fructose-fed streptozotocin-injected rat: an alternative model for type 2 diabetes[J]. Pharmacol Rep, 2012, 64(1): 129-139.

[17] Liu JY, Mu S, Zhang SP, et al. Roux-en-Y gastric bypass surgery suppresses hypothalamic PTP1B protein level and alleviates leptin resistance in obese rats[J]. Exp Ther Med, 2017, 14(3): 2536-2542.

[18] Xu LL, Gao W, Chen ZM, et al. Relationships between diabetic nephropathy and insulin resistance, inflammation, Trx, Txnip, CysC and serum complement levels[J]. Eur Rev Med Pharmacol Sci, 2020, 24(22): 11700-11706.

[19] Zeng JY, Wang Y, Miao M, et al. The effects of rhubarb for the treatment of diabetic nephropathy in animals: a systematic review and meta-analysis[J]. Front Pharmacol, 2021, 12: 602816.

[20] Wang C, Liang J, Yang W, et al. Ultra-performance liquid chromatography-Q-exactive orbitrap-mass spectrometry analysis for metabolic communication between heart and kidney in adriamycin-induced nephropathy rats[J]. Kidney Blood Press Res, 2022, 47(1): 31-42.

[21] Wang C, Hou XX, Rui HL, et al. Artificially cultivated Ophiocordyceps sinensis alleviates diabetic nephropathy and its podocyte injuryvia inhibiting P2X7R expression and NLRP3 inflammasome activation[J]. J Diabetes Res, 2018, 2018: 1390418.

[22] Everaert I, Van der ST, Stautemas J, et al. Oral anserine supplementation does not attenuate type-2 diabetes or diabetic nephropathy in BTBR ob/ob mice[J]. Amino Acids, 2021, 53(8): 1269-1277.

[23] Li Y, Gong W, Liu J, et al. Angiopoietin-like protein 4 promotes hyperlipidemia-induced renal injury by down-regulating the expression of ACTN4[J]. Biochem Biophys Res Commun, 2022, 595: 69-75.

[24] Ladeira LCM, Dos Santos EC, Santos TA, et al. Green tea infusion prevents diabetic nephropathy aggravation in recent-onset type 1 diabetes regardless of glycemic control[J]. J Ethnopharmacol, 2021, 274: 114032.

[25] Chen G, Wang H, Zhang W, et al. Dapagliflozin reduces urinary albumin excretion by downregulating the expression of cAMP, MAPK, and cGMP-PKG signaling pathways associated genes[J]. Genet Test Mol Biomarkers, 2021, 25(10): 627-637.

[26] Qiu Q, Cao J, Wang Y, et al. Time course of the effects of Buxin Yishen Decoction in promoting heart function and inhibiting the progression of renal fibrosis in myocardial infarction caused type 2 cardiorenal syndrome rats[J]. Front Pharmacol, 2019, 10: 1267.

[27] Czirok S, Fang L, Radovits T, et al. Cinaciguat ameliorates glomerular damage by reducing ERK1/2 activity and TGF-β expression in type-1 diabetic rats[J]. Sci Rep, 2017, 7(1): 11218.

[28] Chekalina NI, Burmak YH, Petrov YY, et al. Justification of increasing the blood flow velocity in the arteries of the thyroid gland in autoimmune thyroiditis as a reflection of endothelial changes due to inflammatory status[J]. Curr Med Imaging Rev, 2019, 15(1): 61-65.

[29] Fu Y, Wang X, Zhang L, et al. Allograft inflammatory factor-1 enhances inflammation and oxidative stress via the NF-κB pathway in diabetic kidney disease[J]. Biochem Biophys Res Commun, 2022, 614: 63-69.

[30] Xie X, Shi X, Xun X, et al. Endothelial nitric oxide synthase gene single nucleotide polymorphisms and the risk of hypertension: a meta-analysis involving 63 258 subjects[J]. Clin Exp Hypertens, 2017, 39(2): 175-182.

[31] Wang J, Feng Y, Zhang Y, et al. TNF-α and IL-1β promote renal podocyte injury in T2DM rats by decreasing glomerular VEGF/eNOS expression levels and altering hemodynamic parameters[J]. J Inflamm Res, 2022, 15: 6657-6673.

[32] Harloff M, Prüschenk S, Seifert R, et al. Activation of soluble guanylyl cyclase signalling with cinaciguat improves impaired kidney function in diabetic mice[J]. Br J Pharmacol, 2022, 179(11): 2460-2475.

[33] Ferrini MG, Garcia E, Abraham A, et al. Effect of ginger, Paullinia cupana, Muira puama and L-citrulline, singly or in combination, on modulation of the inducible nitric oxide-NO-cGMP pathway in rat penile smooth muscle cells[J]. Nitric Oxide, 2018, 76: 81-86.

[34] Abdel-Hakeem EA, Abdel Hafez SMN, Kamel BA, et al. Angiotensin 1-7 mitigates rhabdomyolysis induced renal injury in ratsvia modulation of TLR-4/NF-κB/iNOS and Nrf-2/heme-oxygenase-1 signaling pathways[J]. Life Sci, 2022, 303: 120678.

[35] Wu JY, Xie JH, Chen YJ, et al. Amelioration of TPA-induced skin inflammation by the leaf extract of Vernonia amygdalina involves ERK/STAT3 (Ser727) signaling inhibition[J]. Phytomedicine, 2022, 102: 154194.

[36] Assar DH, Asa SA, El-Abasy MA, et al. Aspergillus awamori attenuates ochratoxin A-induced renal and cardiac injuries in rabbits by activating the Nrf2/HO-1 signaling pathway and downregulating IL1β, TNFα and iNOS gene expressions[J]. Environ Sci Pollut Res Int, 2022, 29(46): 69798-69817.

[37] Ma T, Li X, Zhu Y, et al. Excessive activation of Notch signaling in macrophages promote kidney inflammation, fibrosis, and necroptosis[J]. Front Immunol, 2022, 13: 835879.

[38] Slyvka Y, Malgor R, Inman SR, et al. Antioxidant diet and sex interact to regulate NOS isoform expression and glomerular mesangium proliferation in Zucker diabetic rat kidney[J]. Acta Histochem, 2016, 118(2): 183-193.

[39] Xiao S, Li Q, Hu L, et al. Soluble guanylate cyclase stimulators and activators: where are we and where to go[J]. Mini Rev Med Chem, 2019, 19(18): 1544-1557.

[40] Giandalia A, Alibrandi A, Giorgianni L, et al. Resistin levels and inflammatory and endothelial dysfunction markers in obese postmenopausal women with type 2 diabetes mellitus[J]. Diabetol Metab Syndr, 2021, 13(1): 98.

[41] Wennysia IC, Zhao L, Schomber T, et al. Role of soluble guanylyl cyclase in renal afferent and efferent arterioles[J]. Am J Physiol Renal Physiol, 2021, 320(2): F193-F202.

[42] Tzoumas N, Farrah TE, Dhaun N, et al. Established and emerging therapeutic uses of PDE type 5 inhibitors in cardiovascular disease[J]. Br J Pharmacol, 2020, 177(24): 5467-5488.

[43] Mehanna OM, El-Askary A, Al-Shehri S, et al. Effect of phosphodiesterase inhibitors on renal functions and oxidant/antioxidant parameters in streptozocin-induced diabetic rats[J]. Arch Physiol Biochem, 2018, 124(5): 424-429.

[44] Tomita N, Hotta Y, Naiki-Ito A, et al. The phosphodiesterase 5 inhibitor tadalafil has renoprotective effects in a rat model of chronic kidney disease[J]. Physiol Rep, 2020, 8(17): e14556.

[45] Pretorius E. Platelets as potent signaling entities in type 2 diabetes mellitus[J]. Trends Endocrinol Metab, 2019, 30(8): 532-545.

[46] Mao F, Liu S, Qiao X, et al. SUDOSCAN, an effective tool for screening chronic kidney disease in patients with type 2 diabetes[J]. Exp Ther Med, 2017, 14(2): 1343-1350.

[47] Schini M, Peel N, Toronjo-Urquiza L, et al. Evaluation of estimated glomerular function (eGFR) versus creatinine clearance (CrCl) to predict acute kidney injury when using zoledronate for the treatment of osteoporosis[J]. Osteoporos Int, 2022, 33(3): 737-744.

[48] Hong MH, Kim HY, Jang YJ, et al. New therapeutic insight into the effect of Ma Huang Tang on blood pressure and renal dysfunction in the L-NAME-induced hypertension[J]. Evid-Based Compl Alt, 2021, 2021: 9980429.

[49] Piwkowska A. Role of protein kinase G and reactive oxygen species in the regulation of podocyte function in health and disease[J]. J Cell Physiol, 2017, 232(4): 691-697.

[50] Sonneveld R, Hoenderop JG, Isidori AM, et al. Sildenafil prevents podocyte injury via PPAR-γ-mediated TRPC6 inhibition[J]. J Am Soc Nephrol, 2017, 28(5): 1491-1505.

[51] Tomita N, Hotta Y, Naiki-Ito A, et al. Protective effects of tadalafil on damaged podocytes in an adriamycin-induced nephrotic syndrome model[J]. J Pharmacol Sci, 2022, 149(2): 53-59.

[52] Nakamichi R, Hayashi K, Itoh H. Effects of high glucose and lipotoxicity on diabetic podocytes[J]. Nutrients, 2021, 13(1): 241.

[53] Chen Y, Burnett JC. Particulate guanylyl cyclase A/cGMP signaling pathway in the kidney: physiologic and therapeutic indications[J]. Int J Mol Sci, 2018, 19(4): 1006.

Funding

Medical Key Research Projects of Shanxi Province(2020XM02)

Local Science and Technology Development Funds Projects Guided by Central Government(YDZJSX2021C027)

Basic Research Program of Shanxi Province(202103021224370)

Key R & D Project of Shanxi Province [International Scientific and Technological Cooperation, Independent Topics(201903D421061)

Wu Jieping Medical Foundation(320.6750.2021-08-10)