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ORIGINAL ARTICLE
Year : 2021  |  Volume : 9  |  Issue : 3  |  Page : 186-191

Postweaning administration of aqueous leaf extract of Gongronema latifolium may improve obesity indices in young adult offspring


1 Department of Human Physiology, Faculty of Basic Medical Sciences, College of Medical Sciences, Rivers State University, Port Harcourt, Rivers State; Department of Physiology, Faculty of Basic Medical Sciences, Reproductive and Developmental Programming Research Group, College of Medicine, University of Nigeria, Enugu, Nigeria
2 Department of Physiology, Faculty of Basic Medical Sciences, Reproductive and Developmental Programming Research Group, College of Medicine, University of Nigeria, Enugu, Nigeria
3 Department of Physiology, Faculty of Basic Medical Sciences, Reproductive and Developmental Programming Research Group, College of Medicine, University of Nigeria, Enugu; Department of Physiology, Faculty of Basic Medical Sciences, College of Medical Sciences, Alex Ekwueme Federal University Ndufu Alike, Abakaliki, Nigeria
4 Department of Physiology, Faculty of Basic Medical Sciences, Reproductive and Developmental Programming Research Group, College of Medicine, University of Nigeria, Enugu; Physiology Unit, College of Health Sciences, Evangel University, Akaeze Ebonyi State, Nigeria

Date of Submission07-Aug-2021
Date of Decision23-Aug-2021
Date of Acceptance24-Aug-2021
Date of Web Publication30-Nov-2021

Correspondence Address:
Okekem Amadi
Department of Human Physiology, Faculty of Basic Medical Sciences, College of Medical Sciences, Rivers State University, Port Harcourt, Rivers State
Nigeria
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/njecp.njecp_32_21

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  Abstract 


Introduction: Metabolic diseases are multifactorial resulting from genetic, physiological, behavioral, and environmental influences. Genetic influence alone does not suffice to explain the rate at which these diseases have increased. Diet manipulations during critical developmental periods have been used to identify their contribution to obesity and diabetes development in offspring. Gongronema latifolium (GL) has been used for many generations for medicinal and nonmedicinal purposes. The leaves of GL are primarily used as spice and vegetable in traditional folk medicine. Aim: The aim of the study is to investigate the effect of postweaning consumption of aqueous leaf extract of GL on obesity indices in young adult offspring. Materials and Methods: Adult female Wistar rats were used and pregnancy was achieved by introducing matured male Wistar rats of proven fertility at the ratio of two females to one male during proestrus. At the day of delivery, adult female rats were randomly divided into two groups; Group I (normal control) and Groups II–IV (GL extract-treated group). The offspring of the different maternal groups also assumed their mothers' group. Group I was the normal control group while Groups II–IV were given 100, 200, and 400 mg/kg of GL extract, respectively. At postnatal day (PND), 21 offspring were weaned from their mothers and assumed the group of their mothers till PND 42. Parameters such as body weight, body mass index (BMI), waist circumference, body weight-waist circumference ratio, insulin level, liver enzymes (ALT, ALP, and AST), and oral glucose tolerance were assessed in the experimental animals. Results: There was a significant decrease in anthropometric indices (body weight, BMI, waist circumference, body weight-waist circumference ratio) and serum ALT, ALP, and AST levels in the young adult offspring of the GL extract group. However, there was a significant (P < 0.05) increase in insulin level in offspring whose mothers consumed GL extract when compared to the values of the normal control. Conclusions: This study showed that postweaning consumption of GL had significant effects on anthropometric indices, hepatoactivity, insulin sensitivity, and blood glucose level.

Keywords: Anthropometric indices, Gongronema latifolium, insulin, liver enzymes obesity, oral glucose tolerance, sucrose


How to cite this article:
Amadi O, Adeniyi DB, Katchy NA, Nwannadi V, Ugwu PI, Ugwu SU, Iloabachie CR, Emelike CU, Chukwu OO, Iyare CO. Postweaning administration of aqueous leaf extract of Gongronema latifolium may improve obesity indices in young adult offspring. Niger J Exp Clin Biosci 2021;9:186-91

How to cite this URL:
Amadi O, Adeniyi DB, Katchy NA, Nwannadi V, Ugwu PI, Ugwu SU, Iloabachie CR, Emelike CU, Chukwu OO, Iyare CO. Postweaning administration of aqueous leaf extract of Gongronema latifolium may improve obesity indices in young adult offspring. Niger J Exp Clin Biosci [serial online] 2021 [cited 2023 May 29];9:186-91. Available from: https://www.njecbonline.org/text.asp?2021/9/3/186/331558




  Introduction Top


In the last few decades, obesity, metabolic syndrome, and diabetes have escalated to epidemic proportions in many countries worldwide. These metabolic diseases are multifactorial resulting from genetic, physiological, behavioral, and environmental influences. Genetic influence alone does not suffice to explain the rate at which these diseases have increased.[1] In fact, several studies demonstrate that metabolic events during pre- and postnatal development modulate metabolic disease risks in later life.[2] Feeding conditions likely constitute one of the most influential parameters on the health of the adult.[3] Thus, diet manipulation in mothers during critical developmental periods has been used to identify their contribution to obesity and diabetes development in offspring.[4] The importance of early environment, including maternal diet during pregnancy, is suspected to play a major role in the pathogenesis of metabolic syndrome and related conditions. One of the proposed mechanisms is a mismatch between the prenatal and postnatal environments, leading to misprogramming of the metabolic and signaling pathways of the developing fetus.[5],[6]

Plants have been an old companion of man providing food, shelter, wealth and have helped in maintaining relatively good health by its preventive and curative potentials when properly utilized.[7] Medicinal plants constitute an effective source of both orthodox and traditional medicine. Although modern medicine may be available in developing countries, the use of herbs in the treatment of diseases has often gained popularity for historical and cultural reasons. Herbal medicine has been shown to have genuine utility with about 80% of rural dwellers in developing countries depending solely on it for primary health care.[8] The presence of a wide range of bioactive phytochemicals and secondary metabolites has made plants promising source of modern synthetic drugs for the management of several diseases.

Gongronema latifolium (GL) (Asclepiadaceae) is an herbaceous shrub, with yellow flowers and the stem that yields characteristic milky exudates when cut. It is locally called “utasi” by the Efiks, Ibibios, and Quas; “utazi” by the Igbos and “arokeke” by the Yorubas in Nigeria. The Efiks and Quas use GL crude leaf extract in the treatment of malaria, diabetes, hypertension, and as laxative. Furthermore, it is used as a spice and vegetable.[9] The use of crude leaf extract of this shrub in maintaining healthy blood glucose levels has been reported.[10],[11] Scientific studies have established the hypoglycemic, hypolipidemic, and antioxidative effects of aqueous and ethanol extracts of GL leaf.[12],[13] GL has been used for many generations for medicinal and nonmedicinal purposes. The leaves of GL are primarily used as spice and vegetable in traditional folk medicine.[12],[14]

Reports by various authors showed that GL contains essential oils, saponins, and pregnones among others.[9] Ugochukwu and Babady[14] reported that aqueous and ethanolic extract of GL had hypoglycemic effect[9] and showed that it has anti-inflammatory properties. Antilipid peroxidative activity of GL in streptozotocin-induced diabetes was reported by Nwanjo et al.[15] Antibacterial activity has also been reported for GL.[16] Hepatoprotective and hypolipidemic effects have also been reported.[12],[14],[15]


  Materials and Methods Top


Collection of plant material and extraction

Fresh leaves of GL that was used for this study were obtained from local farmers in Nsukka, Enugu state in Nigeria. They were identified and authenticated by a taxonomist at the Department of Plant Science and Biotechnology, University of Nigeria, Nsukka. The method used by Holy et al.[17] was adopted for extraction of the aqueous plant extract. Fresh GL leaves were washed with water, air-dried for 1 week, and homogenized in a warring blender. 1000 g of the finely ground dried plant leaves were immersed into 1000 ml of distilled water. The mixture was allowed to stand for 72 h. It was then filtered using a Whatman No. 1 filter paper. Following filtration, the filtrate was evaporated to dryness over a water bath at 45°C. The pasty filtrate obtained after drying was weighed using an electronic weighing balance. The stock solution of the extract was prepared by dissolving 10 g of the extract in 100 ml of distilled water to give a concentration of 100 mg/ml. The stock solution was appropriately labeled and refrigerated at 4°C until required for use.

Animals

Sixteen adult female Wistar rats (180–250 g) were obtained from the animal house, College of Medicine, University of Nigeria, Enugu Campus and used for this study. The animals were maintained under standard laboratory conditions (12 h light and 12 h dark schedule); they were fed with commercially formulated rat pellets and tap water ad libitum. The animals were allowed to acclimatize to the new laboratory environment for 1 week after which they were divided into groups before the commencement of the experiment.

Experimental design and treatment schedule

The estrus cycle of the female rats was monitored daily under light microscopy. Male Wistar rats were introduced at proestrus at a ratio of two females to a male. The day of appearance of vaginal plug was designated as day 1 of pregnancy. On the day of delivery, the rats were randomly assigned into four groups with four offspring in each group. Group I was the normal control group while Groups II–IV were the extract-treated groups. After day 21, the offspring was separated from their mothers (postweaning period). The offspring of the different maternal groups also assumed their mothers group. Group I was the normal control group while Groups II–IV were given 100, 200, and 400 mg/kg of GL extract, respectively.

Measurement of body mass index

After weaning, the weight and length of the offspring were recorded weekly to postnatal day (PND) 42. The body mass index (BMI) was calculated as the weight of rats (g) divided by the length (cm2). The length of rats was measured between nasal and anal region.[18]

Measurement of waist circumference

Waist circumference of the offspring was recorded (cm) weekly to PND 42.

Measurement of body weight-waist circumference ratio

Body weight-waist circumference was calculated and recorded (g/cm) weekly to PND 42 by dividing the offspring body weight by the waist circumference.

Oral glucose tolerance test

For the oral glucose tolerance test, rats were deprived of food for 12 h. After the food deprivation period, blood samples of the rats were withdrawn from the tail veins. Rats were then given 3 g/kg body weight of glucose solution[19] by oral gavage. The tail blood samples were taken at 0 (just before glucose administration), 30, 60, 90, and 120 min after glucose administration and were analyzed using a glucometer (Accu-Chek), respectively.[20]

Biochemical assay

Serum insulin was measured by enzyme-linked inmunosorbent assay (ELISA) using rat insulin ELISA kit (Mercodia, USA).

Liver function test

Whole blood was separated with high-speed macrocentrifuge at 3500 rev/min for 10 min, and serum was separated by Pasteur pipette for analysis of the following biochemical assays alkaline phosphatase (ALP) as previously described,[21] aspartate aminotransferase (AST),[22] alanine aminotransferase (ALT).[23]

Statistical analysis

Data were expressed as mean ± standard error of mean and statistically evaluated using analysis of variance, followed by a post hoc test using the Statistical Package for the Social Sciences (SPSS) version 23 (IBM SPSS version 23, Chicago, USA). A value of P < 0.05 was considered statistically significant.


  Results Top


[Figure 1] shows the effect of postweaning consumption of GL on offspring body weight. Graded administration of GL (100, 200, and 400 mg/kg) reduced body weight significantly (P < 0.05) when compared with the normal control. 200 mg/kg extract-treated group shows to be more potent in offspring bodyweight reduction.
Figure 1: Body weight of offspring of rats that consumed extract of GL postweaning. I = normal control, II = 100 mg/kg GL, III = 200 mg/kg GL, IV = 400 mg/kg GL. a = P ≤ 0.05 versus control, b = P < 0.05 versus 100 and c = P < 0.05 versus 200. Values expressed as mean ± SEM (n = 4). GL: Gongronema latifolium, SEM: Standard error of mean

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[Table 1] depicts the effect of postweaning consumption of GL on offspring BMI compared to the normal control group, extract treatment significantly (P < 0.05) reduced offspring BMI. Furthermore, 200 mg/kg extract-treated group exerts a more significant (P < 0.05) reduction in offspring BMI than the other extract-treated groups.
Table 1: Body mass index of offspring of rats that consumed extract of Gongronema latifolium postweaning

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As evident in [Figure 2], compared to the normal control, GL treatment significantly (P < 0.05) reduced waist circumference of offspring. [Figure 3] also shows that postweaning consumption of GL significantly (P < 0.05) reduced body weight-waist circumference ratio when compared to the normal control group.
Figure 2: Waist circumference of offspring of rats that consumed extract of GL postweaning. I = normal control, II = 100 mg/kg GL, III = 200 mg/kg GL, IV = 400 mg/kg GL. a = P ≤ 0.05 versus control, b = P < 0.05 versus 100 and c = P < 0.05 versus 200. Values expressed as mean ± SEM (n = 4). GL: Gongronema latifolium, SEM: Standard error of mean

Click here to view
Figure 3: Body weight - waist circumference ratio of offspring of rats that consumed extract of GL postweaning. I = normal control, II = 100 mg/kg GL, III = 200 mg/kg GL, IV = 400 mg/kg GL. a = P ≤ 0.05 versus control, b = P < 0.05 versus 100 and c = P < 0.05 versus 200. Values expressed as mean ± SEM (n = 4). GL: Gongronema latifolium, SEM: Standard error of mean

Click here to view


[Table 2] shows the effect of postweaning consumption of GL on offspring insulin level compared to the normal control, extract treatment significantly (P < 0.05) increased insulin level, with 200 mg/kg being more potent. [Table 3] shows that GL consumption statistically (P < 0.05) decreased liver enzymes (ALT, ALP, and AST) levels when compared to the normal control. [Figure 4] shows that generally postweaning consumption of GL statistically (P < 0.05) decreased oral glucose tolerance when compared to the normal control.
Table 2: Insulin level of offspring of rats that consumed extract of Gongronema latifolium postweaning

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Table 3: Liver function of offspring of rats that consumed extract of Gongronema latifolium postweaning

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Figure 4: Oral glucose tolerance test of offspring of rats that consumed extract of GL postweaning. I = Normal control, II = 100 mg/kg GL, III = 200 mg/kg GL, IV = 400 mg/kg GL. a = P ≤ 0.05 versus control, b = P < 0.05 versus 100 and c = P < 0.05 versus 200. Values expressed as mean ± SEM (n = 4). GL: Gongronema latifolium, SEM: Standard error of mean

Click here to view



  Discussion Top


The present study was undertaken to evaluate the effect of maternal consumption of sucrose during lactation and offspring postweaning consumption of GL on postnatal growth and glucose homeostasis as it pertains to offspring body weight, BMI, waist circumference, body weight-waist circumference ratio, fasting plasma glucose and oral glucose tolerance, insulin level, liver function (ALP, ALT, and AST).

The result showed a decrease in anthropometric parameters (body weight, BMI, waist circumference, and body weight-waist circumference ratio) in the GL extract-treated groups when compared to the normal control group; this may be attributed to the postweaning consumption of GL. This study is consistent with the report of Nwaka et al.[24] This decrease could be due to the presence of the phytochemical saponin[25],[26] or because of the unpalatability of GL due to the bitter substances present in the leaves which could have reduced the rats appetite of the offspring and hence a decrease in the body weight observed in the offspring.[26] The decrease in anthropometric indices is believed to be due activation of adenosine monophosphate (AMP) activated protein kinase (AMPK) by GL polyphenols, saponins that are bioavailable in the liver, skeletal muscle, and adipose tissues. Activated AMPK triggers a decrease in gluconeogenesis and lipogenesis, increases lipolysis, mitochondrial biogenesis (catabolism), and inhibits anabolism,[27],[28],[29] therefore, resulting in a decrease in body weight, BMI, waist circumference. Furthermore, in addition to maintaining cellular energy homeostasis, AMPK responds to different hormone signals to maintain whole-body energy balance.[29]

In this study, the effect of postweaning consumption of GL on offspring insulin level revealed a significant increase in insulin level in GL extract treated groups compared to the normal control. The result reflects a dose-dependent effect of GL extract, this effect demonstrates the antidiabetic potency,[30] as well as blood glucose reducing effect,[31],[32] and insulin stimulating abilities of GL.[33]

Hepatoprotective potential of aqueous leaf extract of GL on hepatic activity in offspring rats was investigated. The extract was administered at the doses of 100, 200, 400 mg/kg body weight orally for 21 consecutive days. The result showed a significant decrease (P < 0.05) in the serum liver enzymes (alanine transaminase [ALT], aspartate transaminase [AST], and alanine phosphatase [ALP]) of all the extract-treated groups compared to normal control group, this is in agreement with the report of Balogun et al.[34] and Imo et al.[35] The normal control showed an increased serum ALT, ALP, and AST values, but this increase may not indicate hepatic damage. ALT is a known hepatospecific enzyme that is principally found in the cytoplasm of hepatocytes.[36],[37] Similarly, AST is present in the cytoplasm and mitochondria of liver cells, and increase in the enzymatic activity of ALT and AST in the serum may directly reflect a major permeability or cell rupture.[38] GL reduces serum liver enzymes, thereby plays a hepatoprotective role and antioxidative effect against oxidative stress and liver damage.[14],[15],[39]

This study also showed that aqueous leaf extracts treatment in rats caused a dose and time-dependent decrease in the blood glucose level compared to the normal control, this is in agreement with the report of Udo et al.[32] The observed hypoglycaemic potency of GL has been suggested by Ugochukwu et al.[12] to be mediated through the activation of hexokinase, phosphofructokinase, glucose-6-phosphate dehydrogenase, and inhibition of glucokinase in the liver. When these enzymes are activated, glycolysis and glycogenesis proceed faster resulting in lowering of blood glucose.[40]


  Conclusion Top


The results of the present study showed that postweaning consumption of GL significantly affected anthropometric indices, hepatoactivity, insulin sensitivity, and blood glucose level.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
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    Figures

  [Figure 1], [Figure 2], [Figure 3], [Figure 4]
 
 
    Tables

  [Table 1], [Table 2], [Table 3]



 

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