ABSTRACT
Background:
The ancient science of life known as Ayurveda uses a variety of formulations or herbal compounds to treat a wide range of illnesses. Male infertility accounts for half of all infertility worldwide, and numerous etiological variables play a role. A pharmacologically active tobacco component called nicotine has a negative impact on male fertility and the reproductive system. Musalyadi Churna (MSC) is one of the polyherbal compound formulations mentioned in Ayurveda Classics for aphrodisiac therapy and recommended for the treatment of male infertility.
Objective:
The current study aims to evaluate the spermatogenic potential of MSC along with its protective effect against testicular degeneration induced by nicotine in rat models.
Materials and Methods:
Sixty adult Sprague–Dawley male rats were used in the experiment, which were divided into ten groups of six rats each. Nicotine was administered orally as disease control. The standard and vehicle control were also included along with study drug groups in which MSC was administered orally to the rats. The study was conducted for 70 days for each group with assessment of body weight and food consumption. After treatment, rats were weighed and sacrificed for evaluation of parameters like organ weight, epididymal sperm count, and sperm morphology along with histological assessment. The data were statistically evaluated using analysis of variance test and Bonferroni multiple-comparison range test to compare different treatment groups.
Results and Conclusion:
In the present study, MSC possesses substantial spermatogenic efficacy, and the result indicates that the study drug possesses a protective effect against nicotine-induced impaired testicular functions in rats, but further research to elucidate its exact mechanism of action is essential.
KEYWORDS: Male infertility, Musalyadi Churna, nicotine, rats, sperm count, testicular degeneration
INTRODUCTION
With the modernization of human civilization, significant changes have occurred in daily life style, food habits, climate, and increased risk of exposure of various toxins. All these factors play a critical role in development of a wide range of diseases, with reproductive system diseases being one of the most serious concerns. Infertility affects approximately 15% of couples worldwide, or 48.5 million couples, with a prevalence of around 23% in India.[1,2] Males have been shown to be solely responsible for 20–30% of infertility cases, accounting for 50% of all cases.[3] Impotency, loss of libido, oligospermia, azoospermia, and necrozospermia are a few increasing predicaments in male. Male infertility is caused in more than 90% of cases by low sperm count (oligospermia) and poor sperm quality, except for some physical defects.[4] Due to their inability to properly achieve pregnancy with their partners, infertile men encounter major psychological and emotional stiga. Nicotine can be consumed in a variety of ways, including smokeless tobacco products such as snuff and chewing tobacco as well as smoked tobacco, which is the most consumed form.[5] Smoking is a lifestyle factor that can contribute to infertility by lowering sperm concentration and quality, which can then cause infertility. It is an extremely hazardous substance that is rapidly absorbed through the oral mucosa, skin, and respiratory tract. Accordingly, it has been reported that nicotine inhibits hormone release from the pituitary gland, adversely affects the hypophysis-gonadal hormone axis, decreases testosterone concentrations in rats, which affects spermatogenesis, significantly reduces semen quality, and also causes degeneration of testicular tissue of rats.[6,7] Many drugs are available from contemporary science to treat oligospermia, but they are associated with some serious adverse effects. Despite great progress in pinpointing the causes, it is still unable to give an appropriate solution. Ayurveda has given the world hope in the treatment of infertility and impotence-related diseases. There are numerous Ayurvedic formulations that are said to have significant Vajikarana properties, which are related to both spermatogenic and spermatogenic aphrodisiac actions.[8] Musalyadi Churna (MSC) is a polyherbal compound formulation in powder form mentioned in a classical Ayurvedic text Bharat Bhaishajya Ratnakar for aphrodisiac therapy and recommended for treatment of erectile dysfunction, premature ejaculation, and oligospermia.[9] It is composed of ingredients such as Shweta Musali (Chlorophytum borivilianum Sant.F.), Vidarikanda (Pueraria tuberosa DC), Guduchi (Tinospora cordifolia (Willd) Miers.), Kapikacchu (Mucunna prurita Hook), Gokshura (Tribulus, terrestris Linn.), Shalmali (Bambax Ceiba DC), Amalaki (Phyllanthus emblica Linn), and Khanda Sharkara (crystalline sugar) in equal proportions. It was suggested that this formulation be consumed orally along with a vehicle like cow’s milk and ghee.[10] Individual drugs in this formulation have been reported to be effective in treating infertility and impotency.[11,12,13,14,15,16,17,18] But there has been no scientific study carried out to assess the spermatogenic properties of this formulation. Hence, the present study was conducted to evaluate the spermatogenic potential of MSC along with its protective effect against testicular degeneration induced by nicotine in Sprague–Dawley rats.
MATERIAL AND METHODS
Experimental animals
The study was carried out after receiving approval from the Institutional Animal Ethical Committee (IAEC approval Research Project No. 10/1920 dated 13/07/2019). Adult Sprague–Dawley male rats weighing 150–250 g, aged 08–10 weeks, were used in the study and supplied by the Animal House of the APT research Center (registration number 40/PO/ReBiRc/S/99/CPCSEA), Pune, India, where also the experimental study was conducted. After acclimatization, rats were housed in an experimental room with a temperature of 22 ± 30°C, a relative humidity of 50–60%, and an illumination cycle of 12 hours dark and 12 hours light. They were fed with solid pellet diet, and potable water that had been filtered through the “Aquaguard” system was always available in plastic bottles with stainless-steel sipper tubes. Food consumption was measured per cage at the start of treatment and every week thereafter. The leftovers from each cage were measured the next day to determine the difference, which was then used to compute the daily food consumption.
Chemicals: Nicotine hydrogen tartrate (95% nicotine) (Siga-Aldrich Chemicals PVT, Bangalore) was used in the study, and its dosage was freshly prepared in normal saline.
Preparation of the test drug
MSC was prepared using traditional methods. Table 1 lists ingredients, parts used, and their proportions. Except for Tinospora cordifolia, all the raw materials used to prepare MSC were purchased from a reliable source and powdered separately before being thoroughly combined. Guduchi Satwa (aqueous starchy extract obtained from Tinospora cordifolia stem, which is solid white powder in appearance) was prepared separately and then combined with other powders to form the final formulation.
Table 1.
Ingredients and quantity of MSC
| Sr. No. | Drug Name | Proportion | Useful parts |
|---|---|---|---|
| 1 | Shweta Musali (Chlorophytum borivilianum Sant. & Fern.) | 1 Part | Rhizome |
| 2 | Amalaki (Phyllanthus emblica Linn.) | 1 Part | Pericarp |
| 3 | Guduchi Satwa (Tinospora cordifolia (Willd) Miers.) | 1 Part | Stem Starchy extract |
| 4 | Vidarikanda (Pueraria tuberosa DC | 1 Part | Rhizome |
| “6 | Shalmali (Bombax Ceiba Linn) | 1 Part” | Root |
| 5 | Gokshura (Tribulus terrestris Linn.) | 1 Part | Fruit, Root |
| 7 | Kapikacchu (Mucunna prurita Hook.) | 1 Part | Seed |
| 8 | Khanda Sharkara (Saccharum officinarum L.) | 1 Part | - |
Selection and preparation of doses: The MSC powder was precisely weighed in the required dose of mg/kg body weight (b.wt.) and then dissolved in 10 ml of a vehicle (equal cow’s milk and cow’s ghee). The dose of MSC stated in the classics is 1 karsha, which is approximately 12 g in the metric system. The dose for this experiment was calculated using the rat conversion factor based on body surface area ratio.[19] Prior to dosing, the weight ofN each individual rat was recorded, and the dosage volume was adjusted accordingly.
Experimental design
The animals were randomly divided into ten experimental groups and treated as shown in Table 2, and each group consisted of six male rats. As the oral route is the clinically intended route for MSC, oral administration was selected in this study. The normal control received distilled water and normal diet for 70 days. Nicotine was administered orally to the disease control group of animals at a dose of 0.6 mg/kg body weight rat. The vehicle control was treated with cow’s milk and ghee (taken in equal proportions) at a dose of 10 ml/kg, and based on the most recent recorded body weights of individual animals, the daily application volume of the vehicle was calculated in advance. The standard control group received nicotine with fertomid. The test drug MSC was tested at three dose levels of human dose, that is, 6 g, 9 g, and 12 g, to reveal the efficacy of the test drug at low dose, intermediate dose, and high dose. Tests 1, 2, and 3 received nicotine with MSC (at 540 mg/kg, 810 mg/kg, and 1080 mg/kg dose levels) and were used to evaluate spermatogenic efficacy of MSC as compared to the standard and disease control. Tests 4, 5, and 6 received only MSC (at 540 mg/kg, 810 mg/kg, and 1080 mg/kg dose levels) and were used to estimate the effect of 70 days MSC treatment on sperm parameters of rats at three dose levels. For all groups, biochemical analyses including serum alkaline phosphatase (ALP), total protein, and serum cholesterol were conducted.
Table 2.
Grouping and posology for spermatogenesis study of MSC in male rats
| Group No. | Groups (n=6) | Dose Specifications | No. of Rats | Route |
|---|---|---|---|---|
| 1 | Normal control (NC) | - | 06 | - |
| 2 | Disease control (DC) | Nicotine (0.6 mg/kg b.wt) | 06 | Oral |
| 3 | Vehicle control (VC) | Cow’s milk and cow’s ghee (10 ml/Kg b.wt | 06 | Oral |
| 4 | Standard drug (STD) | Nicotine (0.6 mg/kg b.wt) + Clomifen citrate (0.6 mg/kg b.wt) | 06 | Oral |
| 6 | Test drug: Low dose (T1) | Nicotine (0.6 mg/kg) + MSC (540 mg/kg b.wt) | 06 | Oral |
| 5 | Test drug: Medium dose (T2) | Nicotine (0.6 mg/kg b.wt + MSC (810 mg/kg b.wt) | 06 | Oral |
| 7 | Test drug: High dose (T3) | Nicotine (0.6 mg/kg) + MSC (1080 mg/kg b.wt) | 06 | Oral |
| 8 | Only test drug (T4) | MSC (540 mg/kg b.wt) | 06 | Oral |
| 9. | Only test drug (T5) | MSC (810 mg/kg b.wt) | 06 | Oral |
| 10. | Only test drug (T6) | MSC (1080 mg/kg b.wt) | 06 | Oral |
i.p: Intraperitoneally, b.wt: Body Weight, MSC: Musalyadi Churna
Epididymal sperm count: The animals were fasted overnight on the last day of the experiment before being sacrificed by cervical dislocation. The epididymal sperm count of the control and treated animals was determined by the method as described in the WHO manual.[20] An incision was made through the cauda of the right epididymis, light pressure was applied to this region, and sperms were extruded. To 1 ml of Ham’s F-10 medium, 10 μl sperms of the control and experimental group were suspended. After centrifugation, 5 μL epididymal sperms were diluted with 95 μL of the diluent (2.5 g sodium bicarbonate, 500 μL of 35% formalin, and 12.5 mg of trypan blue in 50 mL of distilled water) and were counted using a Neubauer hemocytometer. Live sperms appeared bright and colorless, whereas dead sperms showed blue heads.
Sperm Morphology: To assess the percentage of morphologically abnormal sperms, the cauda of the right epididymis was rinsed with 0.5 mL of physiological saline (0.9% NaCl) to obtain a sperm suspension. Aliquots of the sperm suspension were stained with 2% eosin. Hundred spermatozoa per animal were analyzed microscopically at 400× magnification, and we counted spermatozoa with abnormal traits as follows: twisted body, detached head, round tails, and abnormal neck.[21]
Histopathological assessment: Tissues from the testes, liver, and kidney were collected for histopathological examination (2 animals per group.) The organs were exposed under a light microscope after being cut into thin slices using a microtome and stained with hematoxylin–eosin and seen under microscopy (200X) for abnormalities, after which photo-micrographs were taken.
Statistical Analysis: For all treated groups, the results were presented as the mean standard error of mean using SPSS Software (Version 27) and paired t-test, which were used to assess the change in body weight. To compare different treatment groups, the data were statistically evaluated utilizing analysis of variance (ANOVA) and the Bonferroni multiple comparison range test.
OBSERVATION AND RESULTS
Observation and results of Spermatogenesis Study:
DISCUSSION
Herbs produce a diverse range of natural compounds, making them a viable basis for various types of drugs.[22] Musalyadi Churna is a classical polyherbal Ayurvedic formulation chosen for this study because it is used for aphrodisiac therapy and the treatment of various male infertility conditions. Its individual ingredients are widely available, and the formulation is simple to prepare and inexpensive. According to classical texts, the human dose of any powder formulation is approximately 12 g.[23] Ayurvedic physicians, on the other hand, rarely use this dose these days. In general, they recommend a maximum dose of 5 to 6 g for powder formulation. As a result, the MSC dose was chosen to test its efficacy at low doses up to 6 g, medium doses up to 9 g, and high doses up to 12 g.
In the spermatogenesis study, no statistically significant differences were found in body weight and food consumption of normal control and MSC-treated groups and no clinical signs of toxicity were seen during the 70 days treatment period, implying that MSC had no negative effects on the rat’s overall health and metabolism status [Graphs 1 and 2]. Nicotine was selected as a disease control in the present study as it has the ability to reduce reproductive capacity and has a mutagenic effect.[24]
Graph 1.
Body weight comparison from initial to final treatment on Sprague–Dawley rats. NC: Negative Control, VC: Vehicle control, PC: Positive control
Graph 2.
Comparison of food consumption by Sprague–Dawley rats on initial and final treatments during the spermatogenesis study
Organ weight changes are well recognized as a sensitive predictor of chemically induced organ alterations, and a comparison of organ weights between control and treatment groups has traditionally been employed in toxicology investigations to predict the hazardous effects of specific compounds.[25] Clomiphene treatment was reported to induce regressive histological changes in seminal vesicles as well as a considerable weight loss, which was also observed in current study findings.[26] However, as per findings after necropsy and gross examination, the MSC did not cause organ weight reduction or injury to organs, and moreover, it enhances the reproductive organ weight, indicating that the study drug aids spermatogenesis and may also attenuate nicotine’s damaging effects [Graph 3].
Graph 3.
Multiple group comparison graphs of organ weight (gonads and seminal vesicle) of Sprague–Dawley rats
Semen analysis is a crucial factor when evaluating male fertility.[27] The effect of MSC on sperm count, motility, and viability with abnormal or dead sperm data is demonstrated in Table 3 and Figures 1 and 2. The change in sperm parameters could be due to a direct effect on testicular tissue, which results in reproductive dysfunction such as sperm count, motility, and morphology.[28] Graphical representation of live and dead sperm counts in all the treatment groups during the spermatogenesis study is depicted in Graph 4. Comparing disease control rats to normal rats, an 80% reduction in sperm count was observed. This finding supports previous research that suggests nicotine can lower reproductive capacity.[29] Tests 1, 2, and 3 show significant reduction in dead sperm count as compared to the disease control and a non-significant change as compared to the standard group. It indicates that MSC counteracts the sperm damage as caused by nicotine during the treatment period. Only MSC-treated groups (test 4, test 5, and test 6) were found to have a lowered dead sperm count as compared to the normal control. This shows the positive effect of MSC in terms of minimum damage to sperm morphology, which may help to increase the live sperm count and prevent spontaneous sperm damage.
Table 3.
Effects of MSC on sperm counts, motility, and viability of Sprague–Dawley Rats
| Groups | Total sperm counts (million/ml) | Sperm viability (million/ml) | Sperm motility (%) | Abnormal/Dead Sperm (million/ml (%) |
|---|---|---|---|---|
| NC: Normal Control | 25.9 | 20.6±2.6 | 79.54 | 5.3 (20.46) |
| DC: (Nicotine) | 33.1 | 4.1±1.8 | 12.38 | 29.0 (87.61) |
| VC: Vehical Control (Ghee and Milk) | 27.2 | 21.0±1.7 | 77.20 | 6.2 (22.79) |
| STD (Nicotine + Fertomid) | 31.08 | 17.26±3.19 | 57.59 | 13.83 (42.41) |
| Test-1 [Nicotine + MSC (540 mg/kg)] | 29.3 | 12.1±2.4 | 41.29 | 17.2 (58.70) |
| Test-2 [Nicotine + MSC (810 mg/kg)] | 27.8 | 14.8±2.8 | 53.23 | 13.0 (46.76) |
| Test-3 [Nicotine + MSC (1080 mg/kg)] | 29.4 | 9.6±1.5 | 32.65 | 19.8 (67.34) |
| Test-4 [Only MSC (540 mg/kg)] | 24.07 | 20.41±2.58 | 84.79 | 3.66 (15.21) |
| Test-5 [Only MSC (810 mg/kg)] | 26.17 | 21.87±1.09 | 83.57 | 4.3 (16.43) |
| Test-6 [Only MSC (1080 mg/kg)] | 23.4 | 17.7±2.0 | 75.64 | 5.7 (24.34) |
Figure 1.
Images of sperm count and sperm morphology of spermatogenesis study
Figure 2.
Images of normal and abnormal sperms during spermatogenic study
Graph 4.
Spermatogenesis study: Live and dead sperm count
Co-administration of nicotine and MSC (test 1, test 2, and test 3) caused a general and significant improvement in all the sperm parameters (sperm motility, counts, viability, and morphology) when compared to the nicotine group. In the standard group rats (nicotine + fertomid), there was a 76% increase in the sperm count, whereas in test 1 group rats treated with nicotine + MSC (540 mg/kg), there was a 66% increase in the sperm count as compared to the disease control. Further, in test groups 2 [nicotine + MSC (810 mg/kg) and 3 nicotine + MSC (1080 mg/kg)], there were 72.20% and 57.3% increases in sperm count as compared to the disease control. In comparison to the standard control group, the test 2 group showed a non-significant change in sperm counts compared to the test 1 and test 3 groups, indicating that the test 2 group had efficacy similar to the standard control group. Moreover, fertomid (standard drug) does not have additional benefits of an increase in body weight [Graph 1]. The vehicle control group shows a non-significant increase in sperm count as compared to normal.
Sperm morphology test provides a direct measure of the quality of sperm produced in chemically treated animals.[30] Table 4 shows results of sperm morphology of Sprague–Dawley rats in which the nicotine-treated group showed the maximum number of sperm abnormalities in both the head and tail. In MSC-treated groups, a smaller number of dead sperms were observed and normal sperms were found with a better sperm morphology.
Table 4.
Result of Sperm morphology of Sprague–Dawley Rats
| Groups | Sperm Abnormalities | Head-less tail | ||||
|---|---|---|---|---|---|---|
|
| ||||||
| Amorphous Head | Elongated head | Hook less head | Small Tail | Banana Shape Head | ||
| NC: Normal Control | - | - | - | - | - | - |
| DC: (Nicotine) | + | + | + | + | + | + |
| VC: Vehical Control-(Ghee and Milk) | - | - | - | - | - | - |
| STD: (Nicotine + Fertomid) | - | + | + | - | - | - |
| Test-1: [Nicotine + MSC (540 mg/kg)] | - | + | - | - | - | - |
| Test-2 [Nicotine + MSC (810 mg/kg)] | - | - | - | - | - | + |
| Test-3 [Nicotine + MSC (1080 mg/kg)] | + | - | - | - | - | - |
| Test-4 [Only MSC (540 mg/kg)] | - | - | - | - | - | - |
| Test-5 [Only MSC (810 mg/kg)] | - | - | - | - | - | - |
| Test-6 [Only MSC (1080 mg/kg)] | - | - | - | - | - | - |
Serum biochemistry analyses are critical in toxicity studies for determining the potential toxic effects of oral treatment of the test material.[31] Biochemical levels in the standard drug and test were coming toward normal as compared to the disease control [Table 5]. There were non-significant changes observed in MSC-treated groups with comparison to normal by Bonferroni’s multiple comparisons test [Graph 5]. The lack of significant alterations in the levels of ALP, total protein, and serum cholesterol, which are good indicators of liver, kidney, and heart functions, suggests that administration of MSC altered neither hepatocytes and kidneys of rats nor the normal metabolism of the animals.
Table 5.
Results of biochemical estimations of Total protein, ALP, and Cholesterol
| Treatment | Total protein | % Change | ALP | % Change | Cholesterol | % Change |
|---|---|---|---|---|---|---|
| 1. NC | 15.3±1.47 | - | 165.63±16.81 | - | 22.97±2.63 | - |
| 2. DC | 7.86±1 | 48.62 ↓ | 97.37±16.23 | 41.21↓ | 10.16±1.34 | 55.76 ↓ |
| 3. VC | 16±1.35 | 4.57 ↑ | 156.16±14.64 | 5.71 ↓ | 21.83±1.96 | 4.96 ↓ |
| 4. STD | 13.11±0.73 | 66.79 ↑ | 153.83±26.97 | 57.98↑ | 19.77±2.94 | 94.58 ↑ |
| 5. Test-1 | 9.28±1.31 | 18.06 ↑ | 120.92±13.23 | 24.18↑ | 13.44±1.4 | 32.28 ↑ |
| 6. Test-2 | 12.65±1.71 | 60.94 ↑ | 137.53±17.89 | 41.24↑ | 17.44±2.11 | 71.65 ↑ |
| 7. Test-3 | 10.07±1.38 | 28.11 ↑ | 128.28±5.35 | 31.74↑ | 15.99±2.29 | 57.38 ↑ |
| 8. Test-4 | 14.87±1.13 | 2.81 ↓ | 155.75±18.14 | 5.91 ↓ | 24.61±4.71 | 7.13 ↑ |
| 9. Test-5 | 16±1.69 | 4.57 ↑ | 169.2±19.62 | 2.15 ↑ | 20.03±2.97 | 12.79 ↓ |
| 10. Test-6 | 11.91±1.93 | 22.15 ↓ | 157.04±7.6 | 5.18 ↓ | 21.3±2.98 | 7.27 ↓ |
| P | F=3.8884, S, P=0.00093 | F=1.9042, NS, P=0.0733 | F=3.3087, S, P=0.003 |
“Data: Mean±SEM, ↑- Increase, ↓- Decrease, NS- Non Significant, S- Significant, (P<0.05)
Graph 5.
Multiple group comparison graphs of biochemical estimation of total protein, ALP, and cholesterol
Histopathological observations of testes are demonstrated in Figure 3. The normal control group showed the presence of normal seminiferous tubules with intact germinal cell layers. Spermiogenesis with the presence of adequate sperms in lumen of seminiferous tubules was observed. However, due to nicotine treatment for 70 days in the disease control group animals, multi-focal areas of testicular degeneration with loss of germinal cell layers in seminiferous tubules was seen. Focal areas of edematous exudate interstitially between tubules were observed, and there was congestion of vascular tissue, suggestive of spermatic granuloma. There were also minimal signs of reduced spermatogenesis with decreased sperm production in the damaged tubules. Atrophic changes of degenerated and damaged tubules were visible due to nicotine treatment. However, test 1, 2, and 3 groups, in which nicotine was co-administered with MSC at three dose levels, showed normal histomorphological features of testes with intact seminiferous tubules and normal cellular details of seminiferous tubules with various stages of spermatogenesis. This suggests that MSC protects testis tissues from nicotine-induced damage and maintains normal testis functions in these groups. Standard and vehicle control groups also showed normal histomorphological features of testes with intact seminiferous tubules. Test 4, 5, and 6 groups, in which only MSC was administrated, exhibited normal cellular details of seminiferous tubules with various stages of spermatogenesis as well as normal histomorphological aspects of testes with intact seminiferous tubules in both samples. Adequate sperms in lumen of seminiferous tubules were present, indicating spermatogenesis. Histological analysis of liver and kidney tissues showed no abnormalities in MSC-treated groups [Figures 4 and 5]. These findings are supported by normal levels of biochemical estimation values of total protein and serum ALP in MSC-treated groups.
Figure 3.
Representative photomicrographs of histoarchitecture of the testis section (×100)
Figure 4.
Representative photomicrographs of histoarchitecture of the liver section (×100)
Figure 5.
Representative photomicrographs of histoarchitecture of the kidney section (×100)
Due to nicotine treatment, there was a reduction in sperm concentration in the disease control group during the 70-day treatment period, but with MSC, the reduction rate was found to be reduced and the sperm count was significantly increased. This finding demonstrates Musalyadi churna’s spermatogenic potential as the phytoconstituents of MSC may mitigate the deleterious effects of nicotine while also aiding in normal spermatogenesis. The results showed that among the three doses of the study drug, the test 2 group showed maximum efficacy as compared to test 1 and test 3. It may reveal that MSC can be given up to 9 g of human dose for better effects as doses above this level have little impact on sperm count and have a slight negative effect.
MSC ingredients contain a variety of phytochemical[32,33,34] and micro-constituents such as trace elements,[35,36] vitamin C, flavonoids, L-Dopa, and sulfur compounds, which may help in increasing spermatozoa formation and may also stimulate LH and testosterone and further enhance libido, spermatogenesis, and pro-erectile effects. MSC may act on the hypothalamus–pituitary–gonadal axis to improve semen quality. By increasing oxidative defense and maintaining homeostasis in the testicular apoptosis process, MSC may be able to improve the spermatogenesis process. The herbs used in this study may act as immunomodulatory and antioxidant mechanisms, preserving sperm motility and spermatogenesis. As a result of the synergistic action of these ingredients, MSC may be capable of exhibiting highly effective spermatogenic action. Overall, the administration of MSC significantly attenuated the detrimental effect of nicotine on sperm counts, motility, viability, and morphology in rats. This indicates that MSC restored testicular damage induced by nicotine in rats, but further research to elucidate its exact mechanism of action is essential. One limitation of the study is due to a lack of funding, this study could not examine the effects of MSC on reproductive hormones and on other biochemical parameters. The study drug can also be elaborated for its efficacy related to erectile dysfunction in further research.
CONCLUSION
In the present study, pre-clinical evidence is generated for polyherbal compound Musalyadi Churna for possessing substantial spermatogenic efficacy and can be used in the condition of oligospermia. Findings revealed that among the three dose levels, this powder formulation can be given with medium doses for better spermatogenesis effects. According to the results, the study drug protects testis tissues from nicotine-induced damage and maintains normal testis functions in a rat model. Therefore, it is plausible to suggest that Musalyadi Churna can prevent testicular degeneration. However, further research is warranted to elucidate the effect of study drugs, including clinical trials.
Financial support and sponsorship
Technical and financial support from Datta Meghe Institute of Higher Education and Research, DMIHER (DU) University, Wardha.
Conflicts of interest
There are no conflicts of interest.
Acknowledgement
Datta Meghe Institute of Higher Education and Research, DMIHER (DU) University, Wardha is acknowledge for technical and financial support.
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