MOTILIPERM Ameliorates Immobilization Stress-Induced Testicular Dysfunction via Inhibition of Oxidative Stress and Modulation of the Nrf2/HO-1 Pathway in SD Rats
Abstract
:1. Introduction
2. Results
2.1. Body and Organ Weights
2.2. Analysis of Sperm Parameters and Hormone Levels
2.3. Histopathological Analysis and Germ Cell Apoptosis
2.4. Estimation of Malondialdehyde (MDA), ROS/reactive nitrogen species (RNS), and Enzymatic Antioxidant Levels
2.5. Western Blot and Immunohistochemistry Studies of Protein Expression in Testis Tissue
3. Discussion
4. Materials and Methods
4.1. Plant Material and Extract Preparation
4.2. Identification of Major Compounds
4.3. Animal Care and Experimental Design
4.4. Assessment of Sperm Count and Sperm Motility
4.5. Measurement Of Testosterone, Luteinizing Hormone, and Follicle-Stimulating Hormone
4.6. Testicular Histological Studies and Terminal Deoxynucleotidyl Transferase-Mediated (dUTP) Nick-End Labeling (TUNEL) Staining
4.7. Immunohistochemistry Staining
4.8. Malondialdehyde (MDA), Reactive Oxygen Species (ROS)/Reactive Nitrogen Species (RNS), and Antioxidant Enzymes Levels
4.9. Western Blotting
4.10. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
CTR | Control |
M 200 | MOTILIPERM 200 mg/kg |
S | Immobilization stress |
S + M 100 | Immobilization stress + MOTILIPERM 100 mg/kg |
S + M 200 | Immobilization stress + MOTILIPERM 200 mg/kg |
HPLC | High-performance liquid chromatography |
SD | Sprague–Dawley |
HPA | Hypothalamus–pituitary–adrenal |
ER | Endoplasmic reticulum |
Bcl-2 | B-cell lymphoma 2 |
Bax | BCL 2 associated X protein |
StAR | Steroidogenic acute regulatory protein |
MDA | Malondialdehyde |
SOD | Superoxide dismutase |
ROS/RNS | Reactive oxygen species/reactive nitrogen species |
GPx | Glutathione peroxidase |
GPx 4 | Glutathione peroxidase 4 |
Nrf2 | Activating nuclear factor erythroid 2-related factor 2 |
HO-1 | Heme oxygenase 1 |
GnRH | Gonadotropin-releasing hormone |
LH | Luteinizing hormone |
FSH | Follicle stimulating hormone |
TUNEL | Terminal deoxynucleotidyl transferase-mediated (dUTP) nick-end labeling |
H&E | Hematoxylin and eosin |
ST | Seminiferous tubules |
ANOVA | Analysis of variance |
SEM | Standard error of the mean. |
p.o | Per oral |
References
- Guo, Y.; Sun, J.; Li, T.; Zhang, Q.; Bu, S.; Wang, Q.; Lai, D. Melatonin ameliorates restraint stress-induced oxidative stress and apoptosis in testicular cells via NF-kappaB/iNOS and Nrf2/ HO-1 signaling pathway. Sci. Rep. 2017, 7, 9599. [Google Scholar] [CrossRef]
- Vander Borght, M.; Wyns, C. Fertility and infertility: Definition and epidemiology. Clin. Biochem. 2018, 62, 2–10. [Google Scholar] [CrossRef] [PubMed]
- Wang, Y.; Cao, L.; Liu, X. Ghrelin alleviates endoplasmic reticulum stress and inflammation-mediated reproductive dysfunction induced by stress. J. Assist. Reprod. Genet. 2019, 36, 2357–2366. [Google Scholar] [CrossRef]
- Choy, J.T.; Eisenberg, M.L. Male infertility as a window to health. Fertil. Steril. 2018, 110, 810–814. [Google Scholar] [CrossRef] [Green Version]
- Leaver, R.B. Male infertility: An overview of causes and treatment options. Br. J. Nurs. 2016, 25, S35–S40. [Google Scholar] [CrossRef] [PubMed]
- Martins, A.D.; Majzoub, A.; Agawal, A. Metabolic Syndrome and Male Fertility. World J. Mens. Health 2019, 37, 113–127. [Google Scholar] [CrossRef] [PubMed]
- Harlev, A.; Agarwal, A.; Gunes, S.O.; Shetty, A.; du Plessis, S.S. Smoking and Male Infertility: An Evidence-Based Review. World J. Mens. Health 2015, 33, 143–160. [Google Scholar] [CrossRef] [Green Version]
- Ilacqua, A.; Izzo, G.; Emerenziani, G.P.; Baldari, C.; Aversa, A. Lifestyle and fertility: The influence of stress and quality of life on male fertility. Reprod. Biol. Endocrinol. 2018, 16, 115. [Google Scholar] [CrossRef]
- Hess, R.A.; Renato de Franca, L. Spermatogenesis and cycle of the seminiferous epithelium. Adv. Exp. Med. Biol. 2008, 636, 1–15. [Google Scholar] [PubMed]
- Zou, P.; Wang, X.; Yang, W.; Liu, C.; Chen, Q.; Yang, H.; Zhou, N.; Zeng, Y.; Chen, H.; Zhang, G.; et al. Mechanisms of Stress-Induced Spermatogenesis Impairment in Male Rats Following Unpredictable Chronic Mild Stress (uCMS). Int. J. Mol. Sci. 2019, 20, 4470. [Google Scholar] [CrossRef] [Green Version]
- Arun, S.; Burawat, J.; Yannasithinon, S.; Sukhorum, W.; Limpongsa, A.; Iamsaard, S. Phyllanthus emblica leaf extract ameliorates testicular damage in rats with chronic stress. J. Zhejiang Univ. Sci. B 2018, 19, 948–959. [Google Scholar] [CrossRef] [PubMed]
- Arun, S.; Burawat, J.; Sukhorum, W.; Sampannang, A.; Uabundit, N.; Iamsaard, S. Changes of testicular phosphorylated proteins in response to restraint stress in male rats. J. Zhejiang Univ. Sci. B 2016, 17, 21–29. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Mohamadpour, M.; Noorafshan, A.; Karbalay-Doust, S.; Talaei-Khozani, T.; Aliabadi, E. Protective effects of curcumin co-treatment in rats with establishing chronic variable stress on testis and reproductive hormones. Int. J. Reprod. Biomed. (Yazd) 2017, 15, 447–452. [Google Scholar] [CrossRef]
- Bitgul, G.; Tekmen, I.; Keles, D.; Oktay, G. Protective Effects of Resveratrol against Chronic Immobilization Stress on Testis. ISRN Urol. 2013, 2013, 278720. [Google Scholar] [CrossRef]
- Nirupama, M.; Devaki, M.; Nirupama, R.; Yajurvedi, H.N. Chronic intermittent stress-induced alterations in the spermatogenesis and antioxidant status of the testis are irreversible in albino rat. J. Physiol. Biochem. 2013, 69, 59–68. [Google Scholar] [CrossRef]
- Guerriero, G.; Trocchia, S.; Abdel-Gawad, F.K.; Ciarcia, G. Roles of reactive oxygen species in the spermatogenesis regulation. Front. Endocrinol. (Lausanne) 2014, 5, 56. [Google Scholar] [CrossRef] [Green Version]
- Shin, J.S.; Yun, K.J.; Chung, K.S.; Seo, K.H.; Park, H.J.; Cho, Y.W.; Baek, N.I.; Jang, D.; Lee, K.T. Monotropein isolated from the roots of Morinda officinalis ameliorates proinflammatory mediators in RAW 264.7 macrophages and dextran sulfate sodium (DSS)-induced colitis via NF-kappaB inactivation. Food Chem. Toxicol. 2013, 53, 263–271. [Google Scholar] [CrossRef]
- Singh, B.N.; Singh, B.R.; Singh, R.L.; Prakash, D.; Singh, D.P.; Sarma, B.K.; Upadhyay, G.; Singh, H.B. Polyphenolics from various extracts/fractions of red onion (Allium cepa) peel with potent antioxidant and antimutagenic activities. Food Chem. Toxicol. 2009, 47, 1161–1167. [Google Scholar] [CrossRef]
- Soni, K.K.; Zhang, L.T.; Choi, B.R.; Karna, K.K.; You, J.H.; Shin, Y.S.; Lee, S.W.; Kim, C.Y.; Zhao, C.; Chae, H.J.; et al. Protective effect of MOTILIPERM in varicocele-induced oxidative injury in rat testis by activating phosphorylated inositol requiring kinase 1alpha (p-IRE1alpha) and phosphorylated c-Jun N-terminal kinase (p-JNK) pathways. Pharm. Biol. 2018, 56, 94–103. [Google Scholar] [CrossRef]
- Karna, K.K.; Choi, B.R.; You, J.H.; Shin, Y.S.; Cui, W.S.; Lee, S.W.; Kim, J.H.; Kim, C.Y.; Kim, H.K.; Park, J.K. The ameliorative effect of monotropein, astragalin, and spiraeoside on oxidative stress, endoplasmic reticulum stress, and mitochondrial signaling pathway in varicocelized rats. BMC Complement Altern. Med. 2019, 19, 333. [Google Scholar] [CrossRef] [Green Version]
- Zhang, J.H.; Xin, H.L.; Xu, Y.M.; Shen, Y.; He, Y.Q.; Hsien, Y.; Lin, B.; Song, H.T.; Juan, L.; Yang, H.Y.; et al. Morinda officinalis How—A comprehensive review of traditional uses, phytochemistry and pharmacology. J. Ethnopharmacol. 2018, 213, 230–255. [Google Scholar] [CrossRef] [PubMed]
- Banihani, S.A. Testosterone in Males as Enhanced by Onion (Allium Cepa L.). Biomolecules 2019, 9, 75. [Google Scholar] [CrossRef] [Green Version]
- Soni, K.K.; Zhang, L.T.; You, J.H.; Lee, S.W.; Kim, C.Y.; Cui, W.S.; Chae, H.J.; Kim, H.K.; Park, J.K. The effects of MOTILIPERM on cisplatin induced testicular toxicity in Sprague-Dawley rats. Cancer Cell Int. 2015, 15, 121. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Karna, K.K.; Choi, B.R.; You, J.H.; Shin, Y.S.; Soni, K.K.; Cui, W.S.; Lee, S.W.; Kim, C.Y.; Kim, H.K.; Park, J.K. Cross-talk between ER stress and mitochondrial pathway mediated adriamycin-induced testicular toxicity and DA-9401 modulate adriamycin-induced apoptosis in Sprague-Dawley rats. Cancer Cell Int. 2019, 19, 85. [Google Scholar] [CrossRef] [Green Version]
- Soni, K.K.; Shin, Y.S.; Choi, B.R.; Karna, K.K.; Kim, H.K.; Lee, S.W.; Kim, C.Y.; Park, J.K. Protective effect of DA-9401 in finasteride-induced apoptosis in rat testis: Inositol requiring kinase 1 and c-Jun N-terminal kinase pathway. Drug Des. Devel. Ther. 2017, 11, 2969–2979. [Google Scholar] [CrossRef] [Green Version]
- Nargund, V.H. Effects of psychological stress on male fertility. Nat. Rev. Urol. 2015, 12, 373–382. [Google Scholar] [CrossRef] [PubMed]
- Chidrawar, V.; Chitme, H.; Patel, K.; Patel, N.; Racharla, V.; Dhoraji, N.; Vadalia, K. Effects of Cynodon dactylon on Stress-Induced Infertility in Male Rats. J. Young Pharm. 2011, 3, 26–35. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Wagner, H.; Cheng, J.W.; Ko, E.Y. Role of reactive oxygen species in male infertility: An updated review of literature. Arab. J. Urol. 2018, 16, 35–43. [Google Scholar] [CrossRef] [Green Version]
- Mustafa, S.; Wei, Q.; Ennab, W.; Lv, Z.; Nazar, K.; Siyal, F.A.; Rodeni, S.; Kavita, N.M.X.; Shi, F. Resveratrol Ameliorates Testicular Histopathology of Mice Exposed to Restraint Stress. Animals (Basel) 2019, 9, 743. [Google Scholar] [CrossRef] [Green Version]
- Bagheri, Y.; Keshtmand, Z.; Rahbarghazi, R.; Gharamaleki, M.N.; Barati, A.; Bagheri, S.; Rezaie, J.; Rezabakhsh, A.; Ahmadi, M.; Delashoub, M. Salvia officinalis hydroalcoholic extract improved reproduction capacity and behavioral activity in rats exposed to immobilization stress. Anim. Sci. J. 2020, 91, e13382. [Google Scholar] [CrossRef]
- Manna, P.R.; Stetson, C.L.; Slominski, A.T.; Pruitt, K. Role of the steroidogenic acute regulatory protein in health and disease. Endocrine 2016, 51, 7–21. [Google Scholar] [CrossRef] [Green Version]
- Karna, K.K.; Choi, B.R.; Kim, M.J.; Kim, H.K.; Park, J.K. The Effect of Schisandra chinensis Baillon on Cross-Talk between Oxidative Stress, Endoplasmic Reticulum Stress, and Mitochondrial Signaling Pathway in Testes of Varicocele-Induced SD Rat. Int. J. Mol. Sci. 2019, 20, 5785. [Google Scholar] [CrossRef] [Green Version]
- Liu, T.; Wimalasena, J.; Bowen, R.L.; Atwood, C.S. Luteinizing hormone receptor mediates neuronal pregnenolone production via up-regulation of steroidogenic acute regulatory protein expression. J. Neurochem. 2007, 100, 1329–1339. [Google Scholar] [CrossRef]
- Lee, S.H.; Choi, K.H.; Cha, K.M.; Hwang, S.Y.; Park, U.K.; Jeong, M.S.; Hong, J.Y.; Han, C.K.; In, G.; Kopalli, S.R.; et al. Protective effects of Korean Red Ginseng against sub-acute immobilization stress-induced testicular damage in experimental rats. J. Ginseng. Res. 2019, 43, 125–134. [Google Scholar] [CrossRef]
- Gil-Guzman, E.; Ollero, M.; Lopez, M.C.; Sharma, R.K.; Alvarez, J.G.; Thomas, A.J., Jr.; Agarwal, A. Differential production of reactive oxygen species by subsets of human spermatozoa at different stages of maturation. Hum. Reprod. 2001, 16, 1922–1930. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Agarwal, A.; Bui, A.D. Oxidation-reduction potential as a new marker for oxidative stress: Correlation to male infertility. Investig. Clin. Urol. 2017, 58, 385–399. [Google Scholar] [CrossRef] [PubMed]
- Nimse, S.B.; Pal, D. Free radicals, natural antioxidants, and their reaction mechanisms. RSC Adv. 2015, 5, 27986–28006. [Google Scholar] [CrossRef] [Green Version]
- Adewoyin, M.; Ibrahim, M.; Roszaman, R.; Isa, M.L.M.; Alewi, N.A.M.; Rafa, A.A.A.; Anuar, M.N.N. Male Infertility: The Effect of Natural Antioxidants and Phytocompounds on Seminal Oxidative Stress. Diseases 2017, 5, 9. [Google Scholar] [CrossRef] [PubMed]
- Karna, K.K.; Shin, Y.S.; Choi, B.R.; Kim, H.K.; Park, J.K. The Role of Endoplasmic Reticulum Stress Response in Male Reproductive Physiology and Pathology: A Review. World J. Mens. Health. 2019. [Google Scholar] [CrossRef] [Green Version]
- Vomund, S.; Schafer, A.; Parnham, M.J.; Brune, B.; von Knethen, A. Nrf2, the Master Regulator of Anti-Oxidative Responses. Int. J. Mol. Sci. 2017, 18, 2772. [Google Scholar] [CrossRef] [Green Version]
- Loboda, A.; Damulewicz, M.; Pyza, E.; Jozkowicz, A.; Dulak, J. Role of Nrf2/HO-1 system in development, oxidative stress response and diseases: An evolutionarily conserved mechanism. Cell Mol. Life Sci. 2016, 73, 3221–3247. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Ma, Q. Role of nrf2 in oxidative stress and toxicity. Annu. Rev. Pharmacol. Toxicol. 2013, 53, 401–426. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Sahin, K.; Tuzcu, M.; Orhan, C.; Gencoglu, H.; Sahin, N.; Akdemir, F.; Turk, G.; Yilmaz, I.; Juturu, V. MAT, a Novel Polyherbal Aphrodisiac Formulation, Enhances Sexual Function and Nrf2/HO-1 Pathway While Reducing Oxidative Damage in Male Rats. Evid Based Complement Alternat. Med. 2018, 2018, 8521782. [Google Scholar] [CrossRef] [PubMed]
- Bae, W.J.; Ha, U.S.; Choi, J.B.; Kim, K.S.; Kim, S.J.; Cho, H.J.; Hong, S.H.; Lee, J.Y.; Wang, Z.; Hwang, S.Y.; et al. Protective Effect of Decursin Extracted from Angelica gigas in Male Infertility via Nrf2/HO-1 Signaling Pathway. Oxid. Med. Cell Longev. 2016, 2016, 5901098. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Aziz, N.M.; Ragy, M.M.; Gayyed, M.F. Effect of acute immobilization stress with or without a heme oxygenase inducer on testicular structure and function in male albino rats. J. Basic Clin. Physiol. Pharmacol. 2013, 24, 255–262. [Google Scholar] [CrossRef]
- Agarwal, A.; Virk, G.; Ong, C.; du Plessis, S.S. Effect of oxidative stress on male reproduction. World J. Mens. Health 2014, 32, 1–17. [Google Scholar] [CrossRef] [Green Version]
- Niture, S.K.; Kaspar, J.W.; Shen, J.; Jaiswal, A.K. Nrf2 signaling and cell survival. Toxicol. Appl. Pharmacol. 2010, 244, 37–42. [Google Scholar] [CrossRef] [Green Version]
Parameter | CTR | M 200 | S | S + M 100 | S + M 200 |
---|---|---|---|---|---|
Body weight (sacrifice; g) | 400.40 ± 9.15 | 395.90 ± 9.46 | 367.50 ± 5.99 * | 362.33 ± 7.82 | 368.80 ± 3.1 |
Testis weight (g) | 2.03 ± 0.03 | 2.01 ± 0.04 | 1.47 ± 0.15 * | 1.46 ± 0.11 | 1.78 ± 0.07 |
Epididymis weight (g) | 0.62 ± 0.02 | 0.62 ± 0.05 | 0.47 ± 0.02 * | 0.48 ± 0.01 | 0.49 ± 0.01 |
Seminal vesicles weight (g) | 1.66 ± 0.06 | 1.73 ± 0.04 | 1.49 ± 0.05 | 1.49 ± 0.06 | 1.52 ± 0.05 |
Prostate weight (g) | 0.87 ± 0.04 | 0.83 ± 0.02 | 0.78 ± 0.02 | 0.72 ± 0.07 | 0.72 ± 0.03 |
Penis weight (g) | 0.35 ± 0.01 | 0.34 ± 0.01 | 0.35 ± 0.07 | 0.32 ± 0.01 | 0.35 ± 0.01 |
© 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Karna, K.K.; Soni, K.K.; You, J.H.; Choi, N.Y.; Kim, H.K.; Kim, C.Y.; Lee, S.W.; Shin, Y.S.; Park, J.K. MOTILIPERM Ameliorates Immobilization Stress-Induced Testicular Dysfunction via Inhibition of Oxidative Stress and Modulation of the Nrf2/HO-1 Pathway in SD Rats. Int. J. Mol. Sci. 2020, 21, 4750. https://doi.org/10.3390/ijms21134750
Karna KK, Soni KK, You JH, Choi NY, Kim HK, Kim CY, Lee SW, Shin YS, Park JK. MOTILIPERM Ameliorates Immobilization Stress-Induced Testicular Dysfunction via Inhibition of Oxidative Stress and Modulation of the Nrf2/HO-1 Pathway in SD Rats. International Journal of Molecular Sciences. 2020; 21(13):4750. https://doi.org/10.3390/ijms21134750
Chicago/Turabian StyleKarna, Keshab Kumar, Kiran Kumar Soni, Jae Hyung You, Na Young Choi, Hye Kyung Kim, Chul Young Kim, Sung Won Lee, Yu Seob Shin, and Jong Kwan Park. 2020. "MOTILIPERM Ameliorates Immobilization Stress-Induced Testicular Dysfunction via Inhibition of Oxidative Stress and Modulation of the Nrf2/HO-1 Pathway in SD Rats" International Journal of Molecular Sciences 21, no. 13: 4750. https://doi.org/10.3390/ijms21134750