Department of Pharmacognosy and Phytotherapy, Faculty of Pharmaceutical Sciences, University of Port Harcourt, Choba, Rivers State, Nigeria
Introduction: Sorghum is a staple food in Africa and many parts of the world yet with significant medicinal value. This study aims at assessing the anti-fibroid effect of Sorghum bicolor powder in Wistar rats. Methods: Dried seed of Sorghum bicolor was purchased and powdered. Phytochemical screening was conducted on the powder using standard methods. The powder was assessed for acute toxicity in rats. Groups of rats were induced orally with Monosodium glutamate (100 mg/kg) for 30 days and then treated with the powder at 400mg/kg, 800mg/kg and 1200mg/kg for thirty days. Danazol (200mg/kg) was used as standard drug. The weight ratio of the uterus to the weight of the rats, the estradiol, albumin, liver enzymes, kidney and total protein were assessed using standard kits after the rats were sacrificed on the 30th day. Results: Phytochemical screening revealed the presence of alkaloid, triterpenoid, steroids, flavonoids and tannins. Acute toxicity showed Median Lethal Dose (LD50) of 5000mg/kg. The uterine to weight ratio and the estradiol of the treated rats were significantly decreased in a dose dependent manner. No significant effect was observed in the albumin, liver enzymes, kidney and the protein of the treated groups. Conclusion: The Sorghum powder is safe at 5000mg/kg. Biochemical analysis shows reduced level of estradiol and albumin as wells total protein. The study therefore supports its use in combination with other plants in treating uterine fibroid.
Sorghum bicolor L. Monech belongs to the family Poaceae. Sorghum grain has been used traditionally for various purposes across different cultures. Sorghum grain is commonly crushed to make traditional dishes like ugali, which serves as a auxiliary for maize. It is regularly mixed with other grains like cowpeas, amaranthus, and green grams to create nutritious meals.1 Furthermore, sorghum is rich in B-complex vita mins like thiamine and riboflavin, integral to metabolic functions, along with high levels of niacin that improves digestion, skin health, and neurofunction. It also comprises significant amounts of essential minerals, including phosphorous, magnesium, iron, and copper, which contribute to bone health, the production of red blood cells, nerve health, and overall well-being. Additionally, the grain contains some mineral elements in trace quantities including zinc, calcium, and potassium, which offer multiple health benefits. Also noteworthy is sorghum’s unique composition of antioxidants, particularly the 3-deoxy anthocyanins, contributing to various health benefits, including inflammation reduction and potentially decreasing cancer risks. The grain’s diverse nutritional value can, however, depend on factors like its variety, cultivation, and processing techniques applied. For instance, grain sorghum typically has higher protein and lipid levels but less sugar than sweet sorghum.2
The main phenolic acid in sorghum is ferulic. In addition to flavonoids and natural colors, sorghum grain also includes phenolic acids, which are essential for the precise functioning of the enzyme pathway in plant cells. The modified triangular structure of flavonoid molecules includes flavonols (quercetin and kaempferol), flavanones (naringenin), and flavones (apigenin and luteolin).3 Sorghum grain contains ca rotenoids (such as Lutein, Zeaxanthin, and β-Carotene), albeit in quantities that might be lower compared to other grains. In addition, sorghum grain includes antioxidants like α-Tocopherol and α-Tocotrienol, trace amounts of which are measured in micrograms per 100 g of sorghum grain. Sorghum grain also contains polyamines (Spermidine, Spermine, Putrescine, Cadaverine) and phytosterols (β-Sitosterol, Campesterol, Stigmasterol), all of which contribute to its bioactive profile. The presence of these polyamines further contributes to the bioactive profile of sorghum grain. In addition to carotenoids and polyamines, sorghum grain also contains phytosterols, such as β-Sitosterol, Campesterol, and Stigmasterol. The amount of β-Sitosterol is measured in mg/kg, while Campesterol and Stigmasterol are measured in mg/g of lipids. Sorghum grain is also rich in phenolic compounds, including phenolic acids, 3-deoxy anthocyanins, flavonoids, and tannins. Polycosanols are major components of the long-chained lipids isolated from sorghum grain kernels.4
Several biological activities of the secondary metabolites of Sorghum have been reported. These include but not limited to antioxidant activity,5 antidiabetic,6 antimicrobial,7 anti-obesity,8 anti-inflammatory,9 and Anti-atherosclerotic 4 activity. However, there is paucity of information regarding its anticancer potentials but inferences are on the anticancer effect of phenolic compounds detection.10 The terms fibroid, myoma, and leiomyoma are synonymous and are the commonest gynaecological tumors, with an incidence of 60% by the age of 35 and 80% by the age of 50 in African American women.11 It has been observed that local herbal healers make use of Sorghum bicolor in combination with other herbs for the treatment of uterine fibroid. This study aims to verify the effectiveness of the seeds of the plant in the treatment.
MATERIALS AND METHODS
Plant Materials
The seeds of Sorghum bicolor that was used for the investigation purchased from market in Port Harcourt. It was identified and authenticated in the Herbarium of the Department of Pharmacognosy and Phytotherapy, Faculty of Pharmaceutical Sciences, University of Port-Harcourt. The seeds were washed, dried and then pulverized using a milling machine. The powdered form was kept in an airtight container for investigation.
2.2 Experimental Animals
Nonpregnant Seventy healthy albino rats (86g-152g) were acquired from the animal house of the Department of Pharmacology and Experimental Toxicology, Faculty of Pharmaceutical Sciences, University of Port-Harcourt. These rats were acclimatized for one week under standard laboratory conditions.12
Monosodium Glutamate (MSG) induced fibroid
The test animals were grouped into 5 groups, containing 10 rats each. Group 1 receives only 1000 mg/kg of Monosodium Glutamate daily for a period of 30 days. Group 2 receives MSG and 100 mg/kg of standard drug (Danazol). Group 5 receives MSG and powdered Sorghum bicolor of 400 mg/kg, 800 mg/kg and 1200 mg/kg respectively. The animals were given free access to food, water and treatment after eight hours of administration of Monosodium Glutamate.
Determination of Biochemical Parameters
At the end of the 30-day treatment period, blood from the sacrificed rats was collected into plain tubes and allowed to stand for 45 minutes. The clotted samples were centrifuged at 3000 rpm for 15 minutes to separate plasma, which was transferred into sample tubes. Plasma estradiol, total protein, albumin concentration, alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), bilirubin, gamma-glutamyl transferase (GGT), and kidney parameters (creatinine and urea) were determined using Accubind ELISA microwells (Monobind Inc. CA 92630 USA.13
Determination of Uterine/Body Weight Ratio
Weight of animals were determined before induction. After the 30-day treatment period, animals were weighed, before they were sacrificed and dissected to obtain the intact uterus. The uterus was observed, weighed and the uterine/body weight ratio determined.
Statistical analysis
Data obtained from the studies were expressed as mean ± standard error of mean in bar chats. The significance between the mean of the treated and the control animals were established by Student’s T-test
RESULTS
Table 1. Phytochemical screening results
|
S/No |
Phytoconstituents |
Remark |
|
1 |
Alkaloids |
+ |
|
2 |
Triterpenoids |
+ |
|
3 |
Steroids |
+ |
|
4 |
Flavonoids |
+ |
|
5 |
Tannins |
+ |
|
6 |
Anthraquinones |
- |
|
7 |
Cardiac glycosides |
- |
|
8 |
Cyanogenic glycosides |
- |
|
9 |
Saponins |
- |
+ means Present; - means Absent
Table 2. Acute toxicity
|
|
Dose (mg/kg) |
Number of animals |
Number of deaths |
|
|
Phase 1 |
10 |
3 |
0 |
|
|
100 |
3 |
0 |
|
|
|
1000 |
3 |
0 |
|
|
|
Phase 2 |
1600 |
3 |
0 |
|
|
2900 |
3 |
0 |
|
|
|
5000 |
3 |
0 |
|
|
LD50 = 5000mg/kg
Figure 1. Body Weight And Uterine Body Ratio Of the Treated Groups
Figure 2. Estradiol, Total Protein and Albumin Levels Of the Treated Groups
Figure 3. Liver Enzyme Level Of the Treated Groups
Figure 4. Kidney Parameters of the Treated Groups
Figure 5. Electrolyte Levels of the Treated Groups
DISCUSSION
There is a consensus among the scientific community that natural products have been playing a dominant role in the discovery of leads for the development of drugs for the treatment of human diseases.14 In the light of rising health care cost, patients looking for economical approaches to health may consider a combination of herbal and conventional medicine as treatment approaches. Many medicinal plants have been reported to have a positive effect in the treatment of uterine fibroids in Nigerian folklore medicine uses. One of such plants is S. bicolor. The Phytochemical screening showed that the powdered S. bicolor contained Alkaloids, Tannin, Flavonoids, Triterpenes, Carbohydrate while Anthraquinones, Cardiac glycosides, cyanogenic glycosides and saponins were absent (see Table 1). Thus, these substances have a wide spectrum of pharmacological activities of which the most noteworthy include; anti-viral, anti -microbial, anti-cancer, and anti -inflammatory activity.15 In this study, the acute behavioral signs of toxicity such as paw licking, salivation, stretching, and reduced activity, were absent at dose 5000mg/kg although there was a slight increase in the body temperature. There was no mortality at all the dose levels. The oral median lethal dose (LD50) was estimated to be > 5000mg/kg body weight (see Table 2). This high safety profile may have been responsible for its wide spread use as staple food and in ethno-therapeutic interventions.[16]
Uterine to body weight ratio serve as an index in appraising growth within the uterus. Obviously, body weight is expected to increase after 30 days of treatment due to continuous feeding and growth but uterus is expected to decrease due to treatment. The weight ratio of the uterus in Figure 1 shows that the uterus is increasing in size with corresponding increase in the dose of the S. bicolor when compared to the untreated (control) group. The increase in weight of the uterus increases with increase in dose of the powder suggests that the powder is most effective at lower dose. Many compounds have been investigated as potential biomarkers for the diagnosis and surveillance of uterine leiomyomas. Estrogen and progesterone appear to promote the development of myomas, although blood levels of estrogen and progesterone are similar in women with or without clinically detectable myomas, levels of estradiol within myomas are higher than in normal myometrium. Low levels of estradiol to estrone have been found in myomas cells, leading to up-regulation of estrogen, and myoma growth.17 From the Figure 2, the estradiol exhibited a significant maximal decrease at 1200 mg/kg dose of the powder in the treated rats. The protein and albumin level of the groups were insignificantly different from each other. Consequently, the reduction observed in the estradiol of the treated groups did not correspond to their uterine to body weight ratio contrary to Suleiman and Noela.18
The GGT of the treated groups in Figure 3 is significantly lower than the untreated group which suggest that the powder has effect on the GGT of the rats. The bilirubin of the 400 mg/kg treated group was lower than the normal range (0.2 – 0.7 µmol/L) implying reduced antioxidant activity since other groups’ bilirubin are within the normal range. AST and ALP of 400 mg/kg were above the normal range when compared to the untreated (control) and treated with Danazol (standard drug). This appears to be a disturbance since the group with highest dose of the powder has its AST and ALT within the normal range as presented in Figure 3. The blood urea and creatinine are within the normal limit. This suggest that the powder does not have a negative effect on the kidney as presented in Figure 4. The treatment groups in Figure 5, have their electrolytes higher than the normal range as prescribed by Loeb & Quimby.19 The Cl and Na of the treated powder groups are higher than the untreated and treated Danazol groups, thereby suggesting that the powder exhibits other effects than does not have effect on the changes caused by the inducement while K is on normal range. It is observed that there are inconsistencies in the biochemical parameters of the groups treated with the powder of S. bicolor thus, the powder is not used alone in treating fibroid but in combination with other plants which could have some modulation effect on its activity on fibroid.
CONCLUSION
The safety of S. bicolor cannot be over emphasized in this study. However, the powder of S. bicolor could not reduce the weight of the uterus which indicates fibroid treatment but reduce the increment of estradiol. This suggests that S. bicolor is an adjuvant in the combinations of fibroid-herbal preparation.
Ethical matter
This study was done with the approval of Ethic and Research Committee of the University of Port Harcourt.
Conflict of interest
The authors hereby declare that there is no conflict of interest regarding the study and publication of this manuscript.
REFERENCES
Mikailu Suleiman*, Favour Edem Ekpenyong, Evaluation of the Anti Fibroid Potential of the Seeds of Sorghum Bicolor (L.) Monench, Poaceae on Monosodium Glutamate-Induced Uterine Fibroid in Wister Rats, Int. J. of Pharm. Sci., 2025, Vol 3, Issue 5, 183-190. https://doi.org/10.5281/zenodo.15323168
10.5281/zenodo.15323168
