Ameliorative Potentials of Boerhavia diffusa Ethanolic Leaf Extract on cypermethrin-Induced Reproductive Dysfunction in Wistar Rats

Introduction

Boerhaavia diffusa (Family- Nyctaginaceae), is a very controversial plant that has drawn numerous attention across the globe as a result of its immune-modulatory effects [1], anti-lymph and anti-proliferative activity, anti-viral effects [2], as well as its anti-biotic actions. The plant has also been reported to be useful in combating anaemia [3, 4], as a purifier of blood [5], speedy healing of wounds [6], in anti- inflammation [7], etc. Its implications in the quick stoppage of bleeding has also been reported [8], not neglecting its diuretic, cardiotonic [9], antioxidant [10], antihemorrhagic [11], antispasmodic [12], antimicrobial [13], cytotoxic and anticancer roles. Boerhaavia diffusa  is widely dispersed, occurring throughout India, the Pacific, and southern United States. Its flowers are small, around 5 mm in diameter, with round pollens of roughly 65 microns in diameter. This wide range is explained by its small fruit, which are very sticky and grow a few inches off the ground, ideally placed to latch on to small migratory birds as they walk by Hilou A [13].

Although a true and accurate accounting of the native range of  Boerhavia diffusa  has not been determined. However, it is very widespread, and has become  naturalized  in many places.

The root of Boerhavia diffusa acts as an  anticonvulsant,  analgesic [14],  laxative  medication that when rubbed in honey can be locally applied for  cataract, chronic  conjunctivitis  and  blepharitis. Useful for curing heart diseases, anemia and edema (or oedema), Punarnava is an effective remedy that reduces swelling and foul smell in skin disorders. Apart from the root, Punarnava’s leaves are also consumed as a vegetarian dish to reduce oedema. As an Ayurvedic medicine, this herb is said to cure disorders like intestinal colic, kidney disorders, cough, hemorrhoids, skin diseases, alcoholism, insomnia, eye diseases, asthma and jaundice, Boerhavia diffusa (Sri Lankan Name Pita Sudu sarana best green medicene for Diabetes [15].

Recently, a growing number of studies have revealed that reproductive disorders in animals may have a negative impact on pregnancy, from implantation until term. In addition, humans with reproductive disorders often require assisted reproductive technologies to affect pregnancy outcomes. Thus, it is a difficult task to distinguish the contribution of specific reproductive disorders relative to the interventions required for pregnancy success [15, 16]. Also, Hormones and inflammatory mechanisms are implicated in the major events of reproductive functions, while increasing evidence shows that hormonal aberrations and a hyper-inflammatory states may lead to derangements in cross talks that predisposes one to reproductive dysfunction.

Similarly, hematology, which studies the numbers and morphology of the cellular elements of the blood – the red cells (erythrocytes), white cells (leucocytes), and the platelets (thrombocytes) and the use of these results in the diagnosis and monitoring of disease [15, 16] is useful in the diagnosis of many diseases as well as investigation of the extent of damage to blood [17]. Hematological parameters are good indicators of the physiological status of animals. Hematological parameters are those parameters that are related to the blood and blood forming organs.

Results

of a 5ml syringe. Collected samples were then centrifuged at 3000rpm for 15minutes, while refrigerating collected sera for hematological analysis [packed cell volume (PCV), total white blood cell count (TWBCC), total red blood cell (TRBCC) count and total Platelet count (TPC)] at -20°C. Also, animals were dissected to remove the ovaries; which were then cleared of adherent tissues for histological studies.

Discussion

One of the most frequently used group of insecticides are the synthetic pyrethroids. Recently, their continuous use have reportedly caused increase in the levels of chlorines, phosphates and carbamates from organic sources, possibly rapidly degrading, and affecting mammalian toxicity by its action mechanism. Cypermethrin, being a common pyrethroid is useful agriculture and forestry, also in public and animal health fields. Although Cypermethrin has been considered to be nontoxic to mammals, reports have however shown its adverse effects on the neuro-endocrine, renal, hepatic and reproductive systems [16, 17, 18, 19] of laboratory animals; with reproductive toxicity reports posing huge concerns due to susceptibility to chronic exposures. Hence, current study was embarked upon to investigate the reproductive toxicity associated with cypermethrin; and the ameliorative potentials of varying Boerhavia diffusa extract doses in female Wistar rats. Twenty five (25) wistar rats (between 150-200 g) were first procured and acclimatized (two weeks) for this study; following which they were grouped into 5 of n=5 per group. Group I animals were fed standard chow and water at free will (normal control), while inducing reproductive dysfunction in group II rats by oral administration of 3.83 mg/kg bw of cypermethrin for 14 days (negative control). Groups III, IV and V then received (orally) 200 mg/kg bw, 300 mg/kg bw and 400 mg/kg bw of Boerhavia diffusa extract (ethanolic) for five weeks respectively; haven also induced reproductive dysfunction with 3.83 mg/kg bw of cypermethrin.

In this study, Body weight being a valuable indicator of reproductive health was assessed (weekly) across groups (table I). from our result, there was a reduction in body weight of negative control group (II) compared to group I. whereas, administration of varying doses of ethanolic Boerhavia diffusa leaf extract caused a significant (p < 0.05) weekly increase in body weights of all experimental animals compared with control. This increase may portend huge recovery across groups following test administration; and may alter serum sex hormone levels as well (though un-assayed for) to implicate test extract as a possible potent repro-degenerative ameliorator. This finding  is in concordance previous other reports, where cypermethrin treatment was associated with body weight recovery and serum Testosterone, Luteinizing and Follicle Stimulating Hormone levels [20, 21, 22, 23].

Also From table II report of current study, the effect(s) of feeding female wistar rats with varying doses of ethanolic Boerhavia diffusa extract was assayed for selected hematological parameters. From the table, there was a noticeable significant reduction in all assayed hematological parameters for negative control group (II) but for TWBCC which was insignificant compared to control (group I). However, administration of varying doses of ethanolic Boerhavia diffusa leaf extract caused a significant improvement in all variables as compared to control; even though these increases were insignificant across groups for TWBCC as compared with control. Values obtained from the TPC, TWBCC and TRBCC show that B. diffusa ethanolic extract had the most promising effect with increasing doses/ regimen; with remarkable improvements in animals’ physical and behavioural properties as observed.

Again, a clear look at table III reveals a summary of the Histo-Pathological differences observed in the ovaries of Boerhavia diffusa extract treatments to rats at various doses. Here, only moderate number of secondary follicles were noticed in all treatments except group IV; were as, cystic follicles were noticed in Cypermethrin fed, reproductive dysfunctional rats of group II (negative control) as compared to others.

Histologically, this observation showed no noticeable change in the ovaries of treatment groups as against the control and those induced with reproductive dysfunction. However, degenerative follicles of varying sizes were noticed with Hematoxylin and Eosin at a X 400 microscope resolution.

Conclusion

In this study, the ameliorative potentials of ethanolic leaf extract of Boerhavia diffusa was investigated on reproductive and selected hematological variables in wistar rats. From our results, a statistically significant reduction was observed in body weight of 3.83 mg/kg bw of cypermethrin fed rats (reproductive dysfunctional group) as compared to group I. whereas, administration of varying doses of ethanolic Boerhavia diffusa leaf extract caused a significant (p < 0.05) increase in body weights of all experimental animals compared with control group. Also, moderate number of secondary follicles were noticed in all treatments except group IV; were as, cystic follicles were noticed in Cypermethrin fed, reproductive dysfunctional rats. Albeit, histological observations showed no noticeable change in the ovaries of treatment groups as against the control.