Loss of body weight is typically seen to be a harmful impact of the extract on the animal, caused by less consumption of food and liquids. The extract induced a gradual rise in body weight, indicating the extract's relative safety for the rats; yet, there was a noteworthy distinction in the average weekly change in weight acquired (p < 0.05) when compared to the control group (Table 1). In fact, a number of earlier studies have shown that T. tetraptera fruit extract has antiobesity potential [19].

LDL, total cholesterol and triglycerides are three primary components of the lipid profile that are linked to cardiovascular disease. A dysregulated lipid metabolism is indicated by changes in LDL and HDL levels, which may be caused by interference with lipolysis and the release of free fatty acids from peripheral depots [20]. Although the levels of triglycerides, LDL, HDL and total cholesterol were not significantly different (p > 0.05) from the control group, other studies have shown that T. tetraptera fruit extract has the ability to decrease lipids [21]. It has been reported to elicit reduction in serum triacylglycerols as well as elevation of LDL in male rabbits [22].

The primary function of the kidneys is to remove the harmful waste produced by the normal functioning of the body and transported by the blood. In fact, additional research amply supports the capacity of plant extracts to function as potent free radical scavengers in the kidney, avoiding their harmful effects on lipid peroxidation, which raises biochemical markers like creatinine and urea by rupturing membranes [10]. T. tetraptera fruit has been reported to possess alkaloids, tannins, triterpenes, flavonoids, steroid glycosides and coumarins as dominant secondary metabolites [21, 23].

The protective effect of the extract could be explained by the antioxidant potential of some of T. tetraptera extract's constituents, such as flavonoids and total phenols, which would inhibit membrane lysis and contribute to the nephroprotective action of the extract [24]. This study's findings support those of another, which established that some methanol plant extracts protect kidney cells by inhibiting xanthine oxidase, an enzyme that causes lipid peroxidation and membrane instability [25].

The liver is essential for the detoxification and excretion of several endogenous and exogenous substances, and any harm or impairment to it can have a wide range of health effects on both humans and animals. Cellular necrosis, elevated tissue lipid peroxidation and glutathione depletion are linked to liver injury. Furthermore, liver illness is associated with increased serum levels of many biochemical indicators, including transaminases, alkaline phosphatase, triglycerides and cholesterol [26]. All of these parameters, at all tested doses, revealed no significant difference (p > 0.05) compared to the control. These findings suggest that the extract had no effect on the liver's ability to function normally because any hepatocellular damage would have raised ALT and AST levels in the serum [27]. The extract's phenolic components, which function as antioxidants to inhibit membrane lipid peroxidation, may have a hepato-protective effect and could be the reason for this [24].

Significant changes in blood indices (white blood cells, red blood cells, platelets and their differentials) suggest that the chemical being administered is either toxic or protective to the hemopoietic tissue. The blood indices are used to monitor the physiological and pathological state of the body. Findings from our study report nonsignificant effects on most of the important blood indices by the ethanol extract of T. tetraptera fruit. The major functions of WBCs and its differential are to provide immunity and defend the body against invasion by pathogens or toxins. A significant reduction in WBC count at 200 and 400 mg/kg showed that this defense mechanism was not unusually elicited, which proves further the non-toxic nature of the extract. This is consistent with past studies that found that treating white rabbits with an ethanol extract of this plant's fruits reduced the number of white blood cells [22]. Conversely, the dichloromethane methanol extract of this plant resulted in a rise in WBC, which may be explained by the different chemicals that the extraction solvent produced [28]. The extract's nonsignificant effect on the RBC could mean that there was no change in the balance between blood corpuscle destruction to erythropoiesis and the rate of blood production. HGB, PCT and MCH levels did not significantly decrease (p > 0.05) in female rats treated with the various doses; this could indicate that hemoglobin incorporation into red blood cells and red blood cell morphology were unaffected [16]. The dichloromethane methanol extract showed an alteration in this regard.

Toxicity generally happens at the cellular and sub-cellular levels before being seen in tissues. In this study, the low extract dose levels (200 and 400 mg/kg) exhibited little or no significant effect on the histomorphology of the vital organs (Fig. 1). The presence of activated Kupffer cells in the sinusoids at 400 mg/kg is evidence of a boost in the immune system; this can be seen as an extra quality of the extract. Normal architecture observed in the kidney at all doses used shows the level of safety of the extract. This is in tandem with the normal features reportedly observed with the dichloromethane methanol extract of the fruit [28].

The potential benefits of TTF in preventing uterine fibroids were examined in this study. A vital building block for the manufacture of several steroid hormones, which are potent signaling molecules that control a number of bodily processes, is cholesterol [29]. A rise in total serum cholesterol is usually attributed to the activation of the enzyme 3-hydroxyl-3-methoxylglutamyl-CoA reductase (HMGR) which catalyzes the conversion of HMG-CoA to mevalonate; this is the rate-limiting step of cholesterol synthesis [17].

It has been demonstrated that ovarian steroid hormones are important molecular indicators linked to the formation and proliferation of uterine fibroids. The formation and development of uterine fibroids has been attributed primarily to estrogen, but progesterone and its receptors may also have a significant impact [10].

Due to its ability to bind to ERα receptors in the uterus and form a complex that interacts with DNA in the nucleus to activate transcriptional promoter and enhancer regions that govern gene expression, estradiol is unique in its ability to promote the growth of uterine cells. This enables RNA polymerase II binding and the subsequent start of transcription, which results in the production of proteins and higher uterine and ovarian cell proliferation [10].

T. tetraptera fruit extract at graded doses of 100, 200 and 400 mg/kg was evaluated in this study for its ability to inhibit the effect of MSG on these biochemical parameters tested as well as the histology of the uterus because it has been reported to be non-toxic at oral doses as high as 5000 mg/kg [30].

There was a significant (P < 0.05) increase in serum cholesterol levels of the MSG group compared with the normal group (Fig. 2). Treatment with the ethanol extract of T. tetraptera fruit reduced the elevated cholesterol levels almost to normal (P > 0.05) in our study, in both the curative and preventive experiments. The fruit extracts ability to lower cholesterol may be due to a decrease in dephosphorylated HMGR levels as well as an adverse effect on cholesterol production caused by the activation of glucagon and adrenaline [11]. Indeed, the fruit of this plant has been shown in numerous earlier studies to have a lipid-lowering impact [21].

Total protein content in the MSG and treated groups showed no significant difference in relation to the normal. This is similar to results obtained in previous works [7, 17].

Treatment with the ethanol extract of T. tetraptera fruit reduced the elevated estradiol levels in a dose-dependent manner (Fig. 6), in both the curative and preventive experiments. Its action on estradiol could possibly be attributed to suppression of the enzyme aromatase responsible for aromatization of androstenedione and testosterone to estrogens in the biosynthesis of estradiol from cholesterol [4]. It might also be due to an inducer of liver microsomal enzyme that increases the metabolism of estradiol, or it might contain phytochemicals that act as gonadotropin-releasing hormone (GnRH) agonists, which when stimulated continuously reduce the expression or downregulates GnRH receptors on the anterior pituitary [31]. Consequently, less estradiol would be produced. The decline in cholesterol production may potentially also be the cause of the estradiol decline.

Additional histology investigations demonstrated T. tetraptera’s impact on the proliferation of leiomyoma cells in the uterus. A section of the uterus in rats given only food and water revealed normal tissue architecture, but after the female rats were administered 800 mg/kg of MSG, the sections revealed thick bands of spindle-shaped, haphazardly arranged smooth muscle fibers that crisscrossed the endometrial glands and stroma, a characteristic of the formation of leiomyoma.

Gradually increasing dosages of the extract and MSG were administered simultaneously, and the results showed a dose-dependent, stepwise improvement of the proliferating leiomyoma lesion, with the highest dose having the most effective impact. The curative treatment showed a greater reduction in the production of leiomyoma cells than the preventive treatment (Fig. 8).

T. tetraptera fruit has been observed to contain flavonoids, alkaloids, tannins, saponins, steroids, sterols and phenols [23, 28].

Part of the health benefits of saponins include immune system activation and a reduction in cholesterol levels in the body [21]. Additionally, studies have shown that saponin inhibits the enzyme aromatase [14], which is involved in the production of estrogen. Our results are supported by a previous investigation that found a substantial reduction in estrogen induced by a methanol extract of T. tetraptera fruit [32].

Phenols' antioxidant properties are also crucial in preventing chronic diseases because they can shield essential molecules like DNA, lipids and proteins from oxidative damage brought on by reactive oxidant species. While dietary antioxidants can have preventive effects, a low-antioxidant diet can raise the incidence of uterine fibroids [33].

HPLC analysis of the extract of T. tetraptera revealed the presence of major constituents such as umbelliferone, ferulic acid, echinocystic acid, aridanin and naringenin, with hentriacontane, butein and isoliquiritigenin present in reduced amounts (Table 6).

It has been found that the phenylpropanoid umbelliferone possesses antioxidant qualities and effectively inhibits type 3 17β-hydroxysteroid dehydrogenase, which is the main enzyme responsible for converting 4-androstene-3,17-dione into testosterone [34].

Ferulic acid is also a phenolic substance with a variety of biological activities, especially in oxidative stress and inflammation, and also plays an antifibrosis role [35].

Echinocystic acid has been reported to possess antiviral, anti-inflammatory and antioxidation activities [13] while naringenin, a flavanone, has anti-inflammatory, antioxidant and antiproliferative activities [36], and it also showed lipid-lowering properties. Butein, hentriacontane and isoliquiritigenin have numerous pharmacological properties including anti-inflammatory and antioxidative activities [13].

In conclusion, the decreased level of cholesterol and estradiol by effect of T tetraptera fruit extract contributed largely to decreased uterine leiomyoma proliferation. Further studies to determine its exact mechanism as antifibrotic agent against uterine leiomyoma needs to be established.