Protective effect of Ganoderma lucidum spores on epididymis and authophage in diabetic rats

doi:10.12307/2022.851

Abstract

Abstract: BACKGROUND: Hyperglycemia caused by diabetes are closely related to male infertility, and Ganoderma lucidum spores are considered to be anti-aging and improve blood glucose levels. However, the protective effect of Ganoderma lucidum spores on diabetic epididymis remains unclear.
OBJECTIVE: To investigate the protective effects of Ganoderma lucidum spores on diabetic epididymis.
METHODS: Male Sprague-Dawley rats were randomly divided into normal group (n=10), high fat/high glucose group (n=10), and model group (n=20). Rats in the normal group were fed with normal diet, while those in the high fat/high glucose group and model group were fed with high fat and high glucose diet. The rats in the model group were then intraperitoneally injected with streptozotocin (30 mg/kg) to induce diabetes mellitus. Model rats were randomly divided into diabetes mellitus group (n=10) and Ganoderma lucidum spores group (GLS, n=10). The rats in the GLS group were treated with Ganoderma lucidum spores
(300 mg/kg per day) by gavage, and the rats in the high fat/high glucose group and diabetes mellitus group were gavaged with the equal volume of normal saline and fed with high fat and high glucose diet for 12 weeks. Body mass, fasting blood glucose, superoxide dismutase activity, malondialdehyde level and sperm deformity rate were measured in each group. Structural changes in the epididymis in diabetic rats were observed by hematoxylin-eosin staining. The protein expression levels of cleaved caspase-3, beclin1, p62, and LC3 in the rat epididymal tissue were detected by western blot and immunohistochemical staining.
RESULTS AND CONCLUSION: Ganoderma lucidum spores could significantly recovery epididymal function, improve the epididymal histopathology, and promote cell autophagy and antioxidant capacity. Immunohistochemical staining results showed that the immune intensity of p62 and cleaved caspase-3 decreased significantly. Western blot analysis showed that Ganoderma lucidum spores obviously upregulated beclin1 and LC3 protein expression levels and decreased p62 and Cleaved caspase-3 protein expression levels when compared with the diabetes mellitus group. To conclude, Ganoderma lucidum spores may induce autophagy, increase antioxidant capacity and inhibit apoptosis to protect against epididymal injury in diabetic rats.

Key words: diabetes mellitus, epididymis, Ganoderma lucidum spores, high fat and high glucose, autophagy, apoptosis

CLC Number:

Zhang Xin, Ma Xiaoru, Xue Jingwen, Wang Fangfang, Wu Lan, Li Yue. Protective effect of Ganoderma lucidum spores on epididymis and authophage in diabetic rats[J]. Chinese Journal of Tissue Engineering Research, 2022, 26(29): 4632-4637.

Figures/Tables 8

References

[1] HE Z, YIN G, LI QQ, et al. Diabetes Mellitus Causes Male Reproductive Dysfunction: A Review of the Evidence and Mechanisms. In Vivo. 2021; 35(5):2503-2511.
[2] LI Z, SHI Y, ZHANG X, et al. Screening immunoactive compounds of Ganoderma lucidum spores by mass spectrometry molecular networking combined with in vivo zebrafish assays. Front Pharmacol. 2020;11:287.
[3] HYLMAROVA S, STECHOVA K, PAVLINKOVA G, et al. The impact of type 1 diabetes mellitus on male sexual functions and sex hormone levels. Endocr J. 2020;67(1):59-71.
[4] MARESCH CC, STUTE DC, ALVES MG, et al. Diabetes-induced hyperglycemia impairs male reproductive function: a systematic review. Hum Reprod Update. 2018;24(1):86-105.
[5] BOERI L, VENTIMIGLIA E, CAZZANIGA W, et al. Risk of health status worsening in primary infertile men: A prospective 10-year follow-up study. Andrology. 2021. doi: 10.1111/andr.13090.
[6] KOREJO NA, WEI QW, SHAH AH, et al. Effects of concomitant diabetes mellitus and hyperthyroidism on testicular and epididymal histoarchitecture and steroidogenesis in male animals. J Zhejiang Univ Sci B. 2016;17(11):850-863.
[7] CORRÊA LB NS, DA COSTA CAS, RIBAS JAS, et al. Antioxidant action of alpha lipoic acid on the testis and epididymis of diabetic rats: morphological, sperm and immunohistochemical evaluation. Int Braz J Urol. 2019;45(4):815-824.
[8] PEREIRA SC, OLIVEIRA PF, OLIVEIRA SR, et al. Impact of Environmental and Lifestyle Use of Chromium on Male Fertility: Focus on Antioxidant Activity and Oxidative Stress. Antioxidants (Basel). 2021; 10(9):1365.
[9] LIU L, WANG X, LIU K, et al. Inhibition of inducible nitric oxide synthase improved erectile dysfunction in rats with type 1 diabetes. Andrologia. 2021;53(8):e14138.
[10] BARKABI-ZANJANI S, GHORBANZADEH V, ASLANI M, et al. Diabetes mellitus and the impairment of male reproductive function: Possible signaling pathways. Diabetes Metab Syndr. 2020;14(5):1307-1314.
[11] 王东春,肖红,舒琴,等.百令胶囊联合辛伐他汀对2型糖尿病肾病患者氧化与抗氧化失衡及炎性介质水平的影响[J].解放军医药杂志,2020,32(9):39-43.
[12] 张卫华,陈东.叶黄素通过激活Nrf2信号传导通路抑制糖尿病性骨质疏松模型大鼠氧化应激及炎性反应的机制研究[J].临床误诊误治,2020,33(7):99-103.
[13] 杨帆,白抚生,尤林,等.依达拉奉对脑缺血再灌注损伤患者神经功能、氧化应激和炎性反应的影响[J].疑难病杂志,2020,19(9): 880-883+887.
[14] 邓宇飞,翁治委,梁爱军,等.参精固本丸对大鼠睾丸氧化应激损伤后生精功能的修复[J].中华男科学杂志,2019,25(12):1118-1125.
[15] LI D, DING Z, DU K, et al. Reactive Oxygen Species as a Link between Antioxidant Pathways and Autophagy. Oxid Med Cell Longev. 2021; 2021:5583215.
[16] KITADA M, KOYA D. Autophagy in metabolic disease and ageing. Nat Rev Endocrinol. 2021;17(11):647-661.
[17] YUN HR, JO YH, KIM J, et al. Roles of Autophagy in Oxidative Stress.Int J Mol Sci. 2020;21(9):3289.
[18] LI J, LIN FH, ZHU XM, et al. Impact of diabetic hyperglycaemia and insulin therapy on autophagy and impairment in rat epididymis. Andrologia. 2020;52(11):e13889.
[19] WU H, WU S, ZHU Y, et al. Aspirin restores endothelial function by mitigating 17β-estradiol-induced α-SMA accumulation and autophagy inhibition via Vps15 scaffold regulation of Beclin-1 phosphorylation. Life Sci. 2020;259:118383.
[20] 肖博雅,董理鑫,高慧,等.褪黑素可改善PBDE-47所致PC12细胞的异常自噬与凋亡[J].南方医科大学学报,2021,41(9):1409-1414.
[21] PEI C, WANG F, HUANG D, et al. Astragaloside IV Protects from PM2.5-Induced Lung Injury by Regulating Autophagy via Inhibition of PI3K/Akt/mTOR Signaling in vivo and in vitro. J Inflamm Res. 2021;14:4707-4721.
[22] SÁNCHEZ-MARTÍN P, SAITO T, KOMATSU M. p62/SQSTM1: ‘Jack of all trades’ in health and cancer. FEBS J. 2019;286(1):8-23.
[23] DENG Z, LIM J, WANG Q, et al. ALS-FTLD-linked mutations of SQSTM1/p62 disrupt selective autophagy and NFE2L2/NRF2 anti-oxidative stress pathway. Autophagy. 2020;16(5):917-931.
[24] SHIN WH, PARK JH, CHUNG KC. The central regulator p62 between ubiquitin proteasome system and autophagy and its role in the mitophagy and Parkinson’s disease. BMB Rep. 2020;53(1):56-63.
[25] KO JH, YOON SO, LEE HJ, et al. Rapamycin regulates macrophage activation by inhibiting NLRP3 inflammasome-p38 MAPK-NFkappaB pathways in autophagy- and p62-dependent manners. Oncotarget. 2017;8(25):40817-40831.
[26] ZHANG H, ZHANG Y, ZHU X, et al. DEAD Box Protein 5 Inhibits Liver Tumorigenesis by Stimulating Autophagy via Interaction with p62/SQSTM1. Hepatology. 2019;69(3):1046-1063.
[27] MARENINOVA OA, JIA W, GRETLER SR, et al. Transgenic expression of GFP-LC3 perturbs autophagy in exocrine pancreas and acute pancreatitis responses in mice. Autophagy. 2020;16(11):2084-2097.
[28] PUGSLEY HR. Assessing Autophagic Flux by Measuring LC3, p62, and LAMP1 Co-localization Using Multispectral Imaging Flow Cytometry. J Vis Exp. 2017;(125):55637.
[29] LIU WJ, YE L, HUANG WF, et al. p62 links the autophagy pathway and the ubiqutin-proteasome system upon ubiquitinated protein degradation. Cell Mol Biol Lett. 2016;21:29.
[30] JEONG SJ, ZHANG X, RODRIGUEZ-VELEZ A, et al. p62/SQSTM1 and Selective Autophagy in Cardiometabolic Diseases. Antioxid Redox Signal. 2019;31(6):458-471.
[31] BARTOLINI D, DALLAGLIO K, TORQUATO P, et al. Nrf2-p62 autophagy pathway and its response to oxidative stress in hepatocellular carcinoma. Transl Res. 2018;193:54-71.
[32] LIU H, DAI C, FAN Y, et al. From autophagy to mitophagy: the roles of P62 in neurodegenerative diseases. J Bioenerg Biomembr. 2017; 49(5):413-422.