Characterization and Synthesis of Milk Thistle Nanoparticles to overcome Oxidative Stress Induce Testicular damage in male rats


Essa Daham. Jalloob1, Rusul  Heider2, Abdulamir A. Al Zahid3, Iman Jawad. Jaber4, Adnan Mansour. Jasim5, Mohsin6, Rawaa7, Kareem7,

Ameer Ridha. Dirwal7, Dheyaa Ali.Neamah8

1University of Technology / Nanotechnology and Advanced Research Center.

2College of Hilla University/ Department of Anesthetic.

3Al-Safwa University College, Kerbala, Iraq.

4Babylon Veterinary Teaching Hospital.

4Department of Pathology, Veterinary Medicine Collage,  Karbala University.

5,6,7,8College of Veterinary Medicine, AL-Qassim green University Iraq.

*Corresponding Author E-mail:



Background Silymarin (SMN) is a natural component polyphenolic purified from Silybum marianum (milk thistle) seeds. SMN can control reactive oxygen species and converts them into compounds with minimizing their toxicity. Objectives The aim of the present study was to evaluate the therapeutic effect of silymarin nanoparticles on sperm parameters of rats induced diabetes by alloxan. Methods: Firstly gas liquid chromatography (GCMS) was utilized to identification active compound in milk thistle, then the preparation of silymarin -TPGS nanoparticles: Nanoparticles of silymarin of were prepared by the Nano precipitation. Forty Wistar albino rats were divided into four groups, control, induced diabetic with 150mg/kg of alloxan I.p, group three and four induced diabetic and treated with milk thistle extract and nanoparticles at a dose of 100 mg/ daily orally, for 60 days, Blood was collected from heart puncture for estimate serum FSH, L.H, and testosterone, then animal sacrificed to evaluate sperm quality and quantity as well as the histopathological section of tests. Results: Biochemical inspection appear that SMN nanoparticles improved sperm quality and quantity when compared with untreated and SMN groups. SMN nanoparticles appear to have therapeutic effects as an antioxidant against alloxan-induced damage in rat testes. Conclusions: Silymarin nanoparticles can be considered as a hopeful herbal as complementary medicine which may play a significant role to save normal spermatocytes against ROS effects-induced                 reproductive damage.


KEYWORDS: Silymarin, FSH, L.H, Tests, Nanoparticles.




Nanotechnology has been a formed scope that takes action in different fields of medicine, involving cardiology, endocrinology, ophthalmology, immunology, Pulmonology and oncology. In addition, it’s extremely applied in specialized areas like tumor targeting, brain targeting and gene delivery.


Nanotechnology is well efficient to enhancing the bioavailability of water-indissoluble compounds, hold the therapeutic drugs from barriers 20 Literature Review such as blood-brain barrier, highly developed of a novel kind of bioactive polymer as well as load big payloads. Noxious ecological factors, infections, and inflammation lead to reactive oxygen species (ROS) generated by immature sperm cells and leucocytes. cell signaling was dysreulated by ROS, that result defect cellular functions, cells' proliferation, and eventually increase apoptosis1. Oxidative stress and the inflammatory process can influence on sperm and oocyte, which is related with DNA damage or, viability of these cells thus disturbance in motility of sperms that may be aid in development of infertility, abortion, fetus anomaly and sterility2,3. Enzymatic and nonenzymatic antioxidant system save cells against oxidative stress4. Spermatogenesis is complex operation and an extremely concurrent, uniform, long, and that happened in testes at the area of germinal epithelium5. The testes is highly susceptible exogenous and endogenous stress specially in seminiferous tubule. There are wide variety of ROS can influence on tissue testes at different stages of differentiation germinal cells or in somatic testicular cells resulting of temporary or permanent unappealable sterility6. The purification of the Silymarin from milk thistle extract as a hydro-alcoholic form, involving of a compound of the variant of flavonolignans such as silydianin, silychristin, isosilybin A isosilybin B, silybin A and silybin B. These active ingradient involve together around 60–85% of the content of milk thistle extract7,8. Today, silymarin is commonly used as a supportive and alternative therapy in control and relive different medical problems. Awang9 reported that silimarine has a potent antioxidant activity as compared to vitamin E. In addition, Chen et al. notify that silymarin recover Thioacetamide-generate hepatotoxicity10. However, no clear studies have reported the influence of SM on sperm characteristics and fertility. In our project, the influence of silymarin and silymarin nanoparticles on reproductive damage induced by alloxan monohydrate was determined to evaluate sperms parameters.


Characterization and synthesis of Nanoparticles of Silymarin:

Preparation of silymarin -TPGS nanoparticles: Nanoparticles of silymarin of were prepared by the Nano precipitation method according to11,12. Briefly,the organic solution included 100 mg of Silymarin was placed in a glass test tube and then transferred 1ml of acetone solvent, the dissolving Silymarin was incubated overnight, then vortex on high speed until all silymarin extract is entirely dissolved (~10 min). An aqueous phase was prepared by adding 100ml of 0.03% w/v Vitamin E-TPGS to a 200 ml glass beaker for overnight, after that put on a magnetic stir bar at stirring speed 500 rpm, and then 10ml of 0.03% w/v Vitamin E-TPGS was added to glass test tube of emulsified polymer solution. Then the test tube was stirred, then 100μl of silymarine extract was immediately added to the equeous phase of TPGS under high stirring by vortex at 900rpm for 10 min, until all quantity of extract added to TPGS. 5. The emulsified silymarine solution was put in the small beaker 50ml and then directly transferred to the ultrasonicator and immersed in the ice water and sonicate the emulsion for 9min, pulse on time 15 seconds with pulse off time 15 seconds and 50% amplitude. Finally emulsified nanoparticle was filtered (Whatman filter paper 1) to discard any precipitated and then centrifuged at 14,000rpm at 4°C. The supernatant distilled water.


Experimental animals:

The present study was conducted at the, College, of Veterinary Medicine,/Al-Qasim Green University. Adult male rats weighted (180±200g) with ages range between 60-65 days. A total fifty Mature Wister male albino rats with a mean weight of 170 to 190g, obtained from the animal house at the College of Veterinary Medicine, Al Qasim Green University. Diabetes was induced in rats by fasting for more than 16 hours by a intra-peritonealy injection of single dose of 150mg/kg from alloxan monohydrate13.


The animal study groups were divided into four groups according to variation between groups, each group include 10 rats, group control group, group two diabetic rats, group three diabetic rats with silymarin at dosed of 100mg/ daily orally, for 60 day, group four diabetic rats with extract of silymarin nanoparticles at dose 100mg/ daily orally, for 60 day. Blood was collected from heart puncture for estimate serum FSH, L.H and testosterone , then rats were sacrificed by euthanized by dislocation of the cervical spine under anesthesia using a cocktail consist of Ketamine–Xylazine. In the subsequent, step, the tail of the epididymis was digested and macerated, and duct of vas deferens separated into many tiny smaller parts and placed in buffer solution at 37°C for sperm feature calculation.


Sperm motility and Sperm count:

There are different ways used to calculate the movement and the total number of sperm, but we chose the most common procedure has already been described by14 Briefly, for counting sperm, 30μL of the diluted sperm were transmitted to the hemocytometer with a plastic Pasteur pipette and held for 5 min until the sperm reached the counting site. Then, under 400 x magnification, the counting sperms were done in five squares of twenty-five squares the mean was studied. For motility of sperm , briefly, 20μl of the newly ready suspension was lay on the normal slide. Fully five microscopic domains of sperm motility were evaluated and were performed on 200 sperms from each sample. Sperm was counted as "mobile" when it exhibits either progressive movement or spontaneous strikes of the skin if the head of sperm is bind to the vitreous slide. Three separate experiments were outright on more than 100 sperm per case.


Sperm morphology A part of sperm suspension was utilized to make smears to evaluate the sperm shape abnormalities. The morphology of sperm was also planned using of A 20μl of 0.5% nigrosin and eosin then sperms morphology were determined according to assay prescribed by15,16.


Statistical analysis:

The statistical analysis was done by utilizing Complete Randomized Design (CRD( method confirmed by AL-Rawi and Kalaf-Allah17. The mean differences between the averages of the studied traits were fixed at the probability level of (0.01).



Characterization of Silymarin nanoparticles:

The current data regarding polydispersity index (PDI) and particle size of Silymarin nanoparticles with TPGS are presented in figures 1 and 2; they were recorded a effective diameter of 144nm. The size of the NPs was augment by an raise of the by using of aqueous phase TPGS listed in table (1). PDI was employed to determine the distribution of the particle size of nano- suspensions. The results showed a lower PDI at 0.003, which give guide a more homogenous particle size distribution and a stable particle population of uniform distribution. It is a fact that natural products are a healthier match than synthetic drugs. Our result showed that acetone as organic phase and TPGS as aqueous phase reduce particle size and PDI as compared with dimethyl sulfoxide (DMSO) and polyvinyl alcohol (PVA) as surfactant aqueous phase. In addition to acetone is preferable on other organic phase DCM and DMSO , due to easily evaporation from solution due to low melting point18,19. Tween 80 significantly increased drug and Cmax after oral administration, also inhibition of P-gp in the gut improved absorption20. Therefore, Tween 80 is likely to improve systemic exposure of P-gp substrates after oral administration but due to high size 440 nm so that not used in final experiment. In addition to TPGS can act as an effective P-gp inhibitor to hasten uptake of drug by cell as well as increased solubility and stability of therapeutic agents, and their efficacy21,22.


Table(1) Particle size and polydispersity index (PDI), of Silymarin nanoparticles.

Ref. No

Organic phase

Aqueous phase

Particle size nm

Polydispertion index













Tween 80




Table (2): determination absorbance and UV-Visible Spectroscopy of silimarine nanoparticles .

Ref. No



 % T


















λmax (Lambda max), ABS (absorbance), T(transmittance). 


Diagram (1) Refer to particles size nm of silimarine nanoparticles by zeta sizers with polydispersion index


Figure (1): SEM image of particle size and morphology of Silymarin nanoparticles.


Chemical constitute of Silimarine extract:

Table(3) shows that thirteen active ingredients were estimated in silimarine by GC/MS. The main active compounds were Pinene,Terpineol, Caryophyllene, Epiglobulol, Cadinol, Silchristin, Silydianin and Silbin.Kuki, Nagy 23. that confirmed the silimarine active compound like silychristin A, silydianin, silychristin B, silybin A and silybin B, using HPLC chromatograms. Chromatograms of methanolic extracts of dried milk thistle WERE seen eight active compound include , Silychristin, Taxifolin Apigenin-7-glucoside, , Silybin B, Isosilybin A, Silydianin, Silybin A and Isosilybin B, respectively24.


Table 1 Chemical components of extracted clove determined by GC/MS:

Component  retention time

Pinene    5

Terpineol   12

Caryophyllene   18

Epiglobulol   20

Cadinol    21

Silchristin   37

Silydianin   40

Silbin    63


Sperm concentration and sperm motility:

Compared to the results obtained from negative control group (47 X106 ± 1.47and 81±1.68) respectively , the datum confirmed that the average hesitance of Sperm counting and sperm motility a significantly reduced in group 2 at probability, that received alloxan monohydrate at a dose of 150mg/kg to the recorded mean value(23.5 X106±1.55 and 60.25±4.2). Additionally, the rate of group four uncommonly raised than rats in group 3rd. The concentration and motility of sperms appear that the rate of sperm immotile was significantly elevated in group 2 than the 3rd and fourth groups under probability is 0.01. In dissimilarity, a significant elevation in concentration of sperm was spotted when the silimarine nanoparticles when compared with untreated group at probability (0.01), to reach mean value ( 36 x106± 1.58 and 42 x106±2.19).



Figure (2): refer to effect of silimarine nanoparticles on sperm concentration and  sperm motility after exposed rats to alloxan.


The sperm viability significantly reduced while the positive control group received alloxan was compared with the healthy rats at probability 0.01. In contrast, viability of sperm (70±2.4) elevated in the rats of groups 3 that received orally silimarine extract but was statistically significant when compared with fourth group that received silimarine nanoparticles. On the other aspect Sperm abnormality results appear that the mean recurrence of sperm with healthy morphology, a significantly elevated for positive group that received only alloxan when was compared with the negative control group to recorded mean value (45.7±3.37 and 14.25±1.10) respectively at probability 0.01, also, in comparison with the positive group, the mean was significantly minimize after received silimarine in 3th group to recorded mean value (25±3.85). In addition the animal in group fourth that received silimarine nanoparticles showed clear improvement in the percentages of sperm abnormality mimic to negative control group to recorded mean value was (15±1.08).


Effect of nano- silimarine on FSH, L.H and Testosterone:

The present study showed that serum testosterone were significantly reduced in rats received oxidative stress as alloxane monohydrate while the fourth group that received alloxan and treated with nanoparticles of silimarine showed clear improvement to recorded mean value (1±0.09) as compared with the postive control group at (p=0.01). In serum testosterone was still moderate elevation in the rats of groups 3 that received orally silimarine extract but was statistically not significant when compared with positive control group to recorded mean value (0.67±0.03 and 0.38±0.021) respectively. On the other aspect Sperm data were appear that serum FS H and L.H were, significantly elevated for the fourth group that received alloxan and treated with silimarine nanoparticles to recorded mean value (0.75±0.08 and 0.83±0.07) respectively at probability 0.01. In the present study, a significant elevation in concentration sperm was observed in rats received silimarine extract or silimarine nanoparticles. Additionally, alloxan is considered potent oxidative stress could damage testicular tissues such as Sertoli cells and tubules of seminiferous, which, spermatids created in the seminiferous tubule that may drive to developing of the synthesis of spermatogenesis), or produce a defect in the endogenous antioxidant enzymes of epididymis' spermn by the generation of free radical. The finding was in agreement with the study of25. The previous work and reported that rats exposed to alloxan resulted marked reduction in the count of the sperm. Similarly, [26] had noted a marked reduction in the sperm count after exposed to alloxan in rats. Alloxan can affect the expected death of cell by activating caspase 3 which has an essential role in death of cells, and as a consequence, could an important role in sperm reduction.


Figure(3) refer to effect of silimarine extract and silimarine nanoparticles on sperm abnormality and sperm viability after exposed rats to alloxan.


Figure(4) refer to effect of silimarine extract and silimarine nanoparticles on serum level FSH, L.H and Testosterone that exposed rats to alloxan.


The finding was in agreement with the study of25. The previous work and reported that rats exposed to alloxan resulted marked reduction in the count of the sperm. Similarly,26 had noted a marked reduction in the sperm count after exposed to alloxan in rats. Alloxan can affect the expected death of cell by activating caspase3 which has an essential role in death of cells, and as a consequence, could have an important role in sperm reduction. There are several postulated mechanism by which sperm concentration can be adversely affected by alloxan and these mechanisms include direct toxicity of sperm cell or prohibit cell growth thus our data showed clear reduction of testosterone level that effect on sperm concentration and motility. Fatehi27 refer that silymarin at dose 50mg/kg for one week overcome the oxidative stress radiation exposed to rats, through reducing testicular parameters (sperm morphology, concentration, and motility near to normal rats as well as reduce sperm DNA damage with improvement of the testicular tissue. Moreover, administration rats with alloxan monohydrate may induce the mimic influence on the sperm cells and changing functionality of it such as sperm motility and viability, leading to clear defects in the conception operation. Many research reported milk thistle is an herbal potent antioxidant that results in saving cell membranes from lipid peroxidation via reducing the creation of free radicals28.


Regardless of the technique interested, milk thistle has been noted to conserve the genetic material of the sperms from damages, also stimulate cells within tissues to create protein synthesis, and elevate the generation of fresh cells to scavengers the damaged cell, in certain, augment an essential role in many tumors to minimize the adverse effects chemotherapy remedy for normal tissues adjacent tumors. Sperm motility and vitality is the most accurate indicator that assesses the integrity of the sperm's membrane and its ability to conceive29. The mammalian sperm membrane was more susceptible to lipid oxidation in reactive oxygen species causing reduced intracellular ATP and sperm vitality So is sperm motility X Therefore, it is likely that the changes in sperm capacity and motility present in the present study were received by oxidative stress that induces lipid peroxidation in the sperm layer. Since SM contains many active compounds such as silychristin A, silydianin, silychristin B, silybin A and silybin B it has powerful antioxidant properties, which elevate the sperm defense system of antioxidants and support sperm vitality and capacity31. In Additionally, SM has different minerals and vitamines such vitamin C, and E which prevent sperm against alloxan, that saving sperm motility32.


Oufi, Al-Shawi33 reported that silibinin isolated from silymarin lead to a significant elevation in the serum level of testosterone in rapprochement with the negative control group. As well as that to, silibinin increases the diameter of spermatids and primary spermatocytes that improves some testicular functions. on the other aspect, a recent study confirmed that SMN shows a protective effect against methotrexate-induced testicular damage via elevating the activities of antioxidant enzymes34. The SMN remedy preserved from varicocele-induced testicular damage and these rats appears a significant elevation in tubular differentiation, repopulation, and spermiogenesis with the improvement of seminiferous tubules. moreover, SMN clear influence on the varicocele-induced carbohydrate reduction in germinal cells. that has a positive effect on the pathogenesis of varicocele35. The use of antioxidants such as SMN can help restore fertility by scavenging free radicals and blocking transcription factors such as nuclear factor kappa B, with downregulation of E2f1 mRNA.



Figure (5 )  A: rats of animal received alloxan only appear a single tubule with decreased spermatogenic cells and Sertoli cells , The seminiferous tubular atrophy and germinal epitheliums aplasia represent no spermatogenetic activity, the structure of the testes exhibited more disorganization , 40X( H and E). B: rats treated with silimarine extract showed interstitial edema with mild thickening of tunica albugina (H&E; magnification, ×10).


Figure (6) rats treated with nanoparticles of silimarine showed effectively increased the number of germ cells and spermatozoa revealing a typical fully normal appearing spermatogenesis activity in seminiferous tubules(H&E; magnification ×40).



I would like to be grateful University of AL-qassim Green University - College of veterinary medicine – Iraq of its support in the current work.



In conclusion, our findings show that TPGS-coated silymarin nanoparticles have an active phytochemical that can affect the preservation of sperm characteristics after exposure to alloxan which may work by blocking oxidative and inflammatory activity thus increasing the internal antioxidant system or regenerating animals Seminal cells. Accordingly, it may be suitable a complementary herbal medicine to prevent reproductive failure.



This research has been approved by Ethics Commission of Veterinary Medicin according NO:533FD2.



1.      Tahereh, S. and R. Mansoor. Antioxidants and infertility treatment, the role of Satureja Khuzestanica:a mini-systematic review. 2011.

2.      Bisht S et al., Oxidative stress and male infertility. Nature Reviews Urology, 2017. 14(8): p. 470-485.; 69

3.      Sinha A. and S. Gupta. The Role of Antioxidant Supplementation in Male Infertility. Clinical Research in Obstetrics and Gynecology, 2018;1(1): p. 1-12.

4.      Zarif-Yeganeh M. and M. Rastegarpanah. Clinical role of silymarin in oxidative stress and infertility: A short review for pharmacy practitioners. Journal of research in pharmacy practice. 2019; 8(4): p. 181.

5.      Holstein, A.-F., W. Schulze, and M. Davidoff, Understanding spermatogenesis is a prerequisite for treatment. Reproductive Biology and Endocrinology, 2003; 1(1): p. 1-16.

6.      Take G et al. Effect of melatonin and time of administration on irradiation-induced damage to rat testes. Brazilian Journal of Medical and Biological Research, 2009. 42(7):p.621-628.

7.      Surai, P.F., Silymarin as a natural antioxidant: an overview of the current evidence and perspectives. Antioxidants.2015; 4(1): p. 204-247.

8.      Masoumi, H., et al., Clinical and pharmacological applications of silymarin components at cellular and molecular level: a review. Research Journal of Medical Sciences.2016; 10(3): p. 102-107.

9.      Awang D. Milk thistle. Can Pharm J. 1993; 126(4034): p. 422.

10.   Chen I.S., et al., Hepatoprotection of silymarin against thioacetamide‐induced chronic liver fibrosis. Journal of the Science of Food and Agriculture. 2012; 92(7): p. 1441-1447.

11.   Jasim A M. H FHasan, and M J Awady. Preparation of Vorapaxar loaded with Vitamin E TPGS and PVA emulsified PLGA nanoparticles In vitro studies. Research Journal of Pharmacy and Technology. 2019; 12(9): p. 4503-4510.

12.   Jasim A M et al., Characterization and Synthesis of Selenium-TPGS Nanoparticles for Target Delivery Clove to Minimize Cytogenic and Liver Damage Induced in Adult Male Rats. Nano Biomed. Eng, 2021. 13(2): p. 127-136.

13.   Jasim A A Dirwal and AAl-Yassri. Role of zontivity loaded by PLGA coated by TPGS in ameliorating induced cardiovascular and pulmonary disorder in adult male. East African Medical Journal, 2019; 96(6).

14.   CooperT G., et al. World Health Organization reference values for human semen characteristics. Human reproduction update, 2010; 16(3): p. 231-245.

15.   Adnan, M., et al., Effect of long-term administration of Atonik compound in female rabbits on hematological and pathological changes in important functional body organ. Journal of Contemporary Medical Sciences, 2016. 2(7): p. 96-102.

16.   Ibraheem AK. efficacy of dq-tocopheryl polyethylene glycol succinate (tpgs) to minimize atonik toxicity in male rats. Plant Archives. 2020; 20(1): p. 1738-1342.

17.   Al-Rawi, K. and A. Khalafalla, Analysis of Experimental Agriculture Disgen. Dar Al-Kutub for Printing and Publishing. Mosul Univ. 2000.

18.   Sah, E. and H. Sah, Recent trends in preparation of poly (lactide-co-glycolide) nanoparticles by mixing polymeric organic solution with antisolvent. Journal of Nanomaterials, 2015; 2015.

19.   Dibenedetto A., et al. Organic carbonates: efficient extraction solvents for the synthesis of HMF in aqueous media with cerium phosphates as catalysts. ChemSusChem, 2016; 9(1): p. 118-125.

20.   Zhang H et al., Commonly used surfactant, Tween 80, improves absorption of P-glycoprotein substrate, digoxin, in rats. Archives of pharmacal research, 2003;26(9)

21.   Guo Y . The applications of Vitamin E TPGS in drug delivery. European journal of pharmaceutical sciences, 2013. 49(2): p. 175-186.

22.   Mohammed QA. protective role of vitamin–tpgs to overcome oxidative stress induced by dipping of sheep with cypermethrin. Plant Archives, 2020; 20(1): p. 1105-1109.

23.   Kuki, A., et al., Identification of silymarin constituents: an improved HPLC–MS method. Chromatographia, 2012. 75(3-4): p. 175-180.

24.   Tayoub G. H Sulaiman, and M ALorfi. Quantitative identification of total silymarin in wild Silybum marianum L. by using HPLC. International Journal of Herbal Medicine, 2018;6(2): p. 110-114.

25.   Serban C. Effect of sour tea (Hibiscus sabdariffa L.) on arterial hypertension: a systematic review and meta-analysis of randomized controlled trials. Journal of hypertension, 2015; 33(6): p. 1119-1127.

26.   Elangovan A et al. Momordica cymbalaria improves reproductive parameters in alloxan-induced male diabetic rats. 3 Biotech, 2021;11(2): p. 1-14.

27.   Fatehi D et al. Radioprotective effects of Silymarin on the sperm parameters of NMRI mice irradiated with γ-rays. Journal of Photochemistry and Photobiology B: Biology, 2018; 178: p. 489-495.

28.   Adhikari M et al. In vitro studies on radioprotective efficacy of silymarin against γ-irradiation. International journal of radiation biology, 2013; 89(3): p. 200-211.

29.   Wong A et al. Addition of eosin to the aniline blue assay to enhance detection of immature sperm histones. Fertility and sterility, 2008;90(5): p. 1999-2002.

30.   Fanoudi S et al. Milk thistle (Silybum Marianum) as an antidote or a protective agent against natural or chemical toxicities: a review. Drug and chemical toxicology, 2020;43(3): p. 240-254.

31.   Tuli H S et al. Path of Silibinin from diet to medicine: A dietary polyphenolic flavonoid having potential anti-cancer therapeutic significance. in Seminars in Cancer Biology. 2020; Elsevier.

32.   Bansal, J., et al., Hepatoprotective Models and Various Natural Product Used in Hepatoprotective Agents: a Review. Pharmacognosy Communications, 2014; 4(3).

33.   Oufi H G. N N. Al-Shawi, and S.A. Hussain, What are the effects of silibinin on testicular tissue of mice? Journal of Applied Pharmaceutical Science, 2012; 2(11): p. 9.

34.   Yaman T. Protective effects of silymarin on methotrexate-induced damages in rat testes. Brazilian Journal of Pharmaceutical Sciences, 2018; 54(1).

35.   Moshtaghion S M . Silymarin protects from varicocele-induced damages in testis and improves sperm quality: evidence for E2f1 involvement. Systems biology in reproductive medicine, 2013; 59(5): p. 270-280.




Received on 28.04.2021            Modified on 08.09.2021

Accepted on 04.11.2021           © RJPT All right reserved

Research J. Pharm.and Tech 2022; 15(4):1664-1670.

DOI: 10.52711/0974-360X.2022.00278