Recently use of herbal therapies and diet rich in flavonoids and vitamins has augmented significantly to manage minor to modest illnesses. Several anti-hyperlipidemic agents are currently available; however most of them have various unwanted effects. That’s why more people are switching towards safer alternatives, specially derived from plants with fewer side effects. Hence current study was focused to determine the anti-hyperlipidemic potential of Nelumbo nucifera fruit in rabbits fed with high cholesterol diet. This study would surely help in the exploration of a novel agent from the plant source and therefore may recommend the application of traditional medicines in humans. 28 white healthy rabbits of either sex weighing 1000 - 1500 gm were divided into four groups. Control group was given gum tragacanth and 3 groups served as test groups and were given N. nucifera fruit extract 50, 100 and 200 mg/kg for 45 days. Initially animals of all four groups received high cholesterol diet (HCD) 0.125 gm/kg cholesterol in 0.5% corn oil for 30 days. All drugs were given orally. Blood samples were taken thrice from the ear vein of animals, 1 st after 24 hours of thirty days of HCD then again after 24 hours of thirty and forty five days dosing of N. nucifera fruit extract. The results showed remarkable cholesterol lowering effects of N. nucifera fruit extract at doses 50, 100 and 200 mg/kg but were particularly dominant at 200 mg/kg at which fruit extract highly significantly reduced TC, TG and LDL-C after 30 and 45 days and increased HDL-C significantly after 30 days and highly significantly after 45 days. Flavonoids, saponins, tannins and procyanidin are significant constituents of N. nucifera fruit which may have contributed in lipid lowering effects of N. nucifera fruit which can contribute in lowering the risk of CVDs and stroke but more preclinical and clinical studies are required to confirm these findings.
Herbal Medicine or Herbalism is the practice or art of employing herbs and herbal preparations in order to remain healthy and also for the treatment and improvement in prognosis of diseases [
These days vigorous research is ongoing to discover nontoxic and beneficial herbs. Several anti-hyperlipidemic agents are currently available; however most of them have associations with various unwanted effects. That’s why more people are switching towards safer alternatives, specially derived from plants with fewer side effects [
N. nucifera belongs to family Nymphaeaceae, and is frequently named as bean of India, Chinese water lily, Indian lotus or merely lotus. According to Linnaean classification N. nucifera Gaertn is at present documented name of plant species and is widely cultivated in the tropical regions of Pakistan, India, China, Thailand and Australia [
N. nucifera fruit is commonly known as Paban in native language of Sindh province of Pakistan and is cultivated especially in the water gardens, ponds and lakes of interior Sindh (Gotki, Thatta, Larkana and Matiari districts) and in Muzaffergarh regions of southern Punjab, Pakistan. It is also found abundantly in the lakes and streams of Azad Jammu Kashmir (AJK), Pakistan.
Fruit consists of seeds and seed pods. The latter is also known as lotus bulbs, are green in color and provides attachment to the seeds, which are black in color, hard in nature, ovoid, oblongish or roundish in shape, up to 1.5 cm broad and 1.0 cm long and are organized in whorls [
The N. nucifera seeds are loaded with fat, amino acid, protein, asparagines, unsaturated fatty acids, minerals, starch, saponins and tannins. The seed has three main components; cotyledons, plumule and integuments, which occupies approximately 93.23%, 3.03% & 3.74% in terms of its mass correspondingly. The average burden of hundred seeds is approximately 87.35 g. The seeds also contain substantial percentage of various minerals such as calcium (22.10%), potassium (28.5%), magnesium (9.2%), sodium (1.0%), iron (0.19%), chromium (0.004%), manganese (0.35%), zinc (0.08%) and copper (0.046%). Other significant dietary elements include fat (72.17%), proteins (2.7%), total powdery residue (4.5%), wetness (10.50%) and raw fiber (10.60%). The seed are highly nutritious and possesses 348.45 cal per 100 g [
The qualitative phytochemical analysis of the seed pods of N. nucifera fruit exposed the presence of flavonoids, alkaloids, saponins, tannins and terpenoids [
Traditionally fruits of N. nucifera are used in enormous conditions such as fever, inflammation, insomnia, palpitation, hypertension, arrhythmia, enteritis, chronic diarrhea, spermatorrhea, leucorrhoea, halitosis, skin diseases, leprosy and menorrhagia. It is also used as an antiemetic, antidote, refrigerant and diuretic [
Experimental design & methodology
Research work was executed utilizing the laboratory facilities of Pharmacology department and the Research Institute of Pharmaceutical Sciences, University of Karachi, following approval from the Board of Advance Studies & Research (BASR), and departmental research & ethical committee.
Anti-hyperlipidemic effect was evaluated in 28 healthy white rabbits of either sex weighing 1000 - 1500 gm which was divided in to four groups; control and 3 test groups. 7 rabbits were placed in each group. Rabbits were chosen as experimental animals for current study as biochemical changes produced in rabbits are very much similar as observed in humans [
Animals were placed in plastic cages with preservation of room temperature at 23˚C ± 2˚C and moisture kept at 50% to 60% in a substituting 12-h light/dark succession. Every animal was provided with normal diet prepared in laboratory plus water as desired. The animals were shifted to the laboratory approximately an hour before the commencement of experiments. The experiment was performed during day time. Prior to dose administration, overall fitness of animals were assessed throughout the adaptation phase utilizing the laboratory conditions for a week particularly observing lack of activity, edema, diarrhea and ulceration.
The research committee Department of Pharmacology permitted the use of animals for these experiments in agreement with the guidelines of NIH [
N. nucifera fruits were obtained from the local fruit & vegetable market of Qasimabad, Hyderabad, Sindh, Pakistan. The N. nucifera fruits were identified and authenticated in the Department of Pharmacognosy. The receipt sample no NNF-03 was placed in the section of Pharmacognosy, University of Karachi.
Grounding of crude extract followed cold extraction procedure [
The solvent was filtered with the help of Whatman No. 1 filter paper. Afterwards the extract was evaporated under reduced pressure in a rotary evaporator at 40˚C to 45˚C followed by freeze drying at −30˚C. The solid Lyophilized material so obtained was saved at −20˚C until further use in doses of 50, 100 and 200 mg/kg orally [
0.5% Corn oil was purchased from local medical stores in Karachi, while gum tragacanth and cholesterol powder were supplied by Merck. High cholesterol diet (HCD) i.e. 0.125 gm/kg cholesterol in 0.5% corn oil was given to rabbits for 30 days in order to derange lipid profile [
The suspending agent (gum tragacanth powder) was consumed to prepare suspensions of the control and 3 doses of test group (N. nucifera fruit extract 50, 100 & 200 mg/kg). It was administered to control group as placebo in the dose of 10 ml/kg per oral, every time fresh suspensions were made for the purpose of dosing [
A population of 28 white healthy rabbits of either sex was divided in to four groups. Control group was given gum tragacanth and 3 groups served as test groups and were given N. nucifera fruit extract 50, 100 and 200 mg/kg for 45 days. Initially animals of all four groups received high cholesterol diet (HCD) i.e. 0.125 gm/kg cholesterol in 0.5% corn oil for 30 days. All agents were given PO [
Blood samples (5 ml) were collected in gel tube. Serum was instantly separated by centrifuging blood samples on 14 K Humax centrifuge at 3000 rpm for 15 min. Lipid profile was investigated on Humalyzer 3000 (semi-automatic chemistry analyzer, Model #16700) (Human Germany) using standard kits supplied by Human. TC and LDL-C was assessed by CHOD-PAP method; TG by GPO-PAP method [
The data was subjected to analysis by taking average and standard error to the average utilizing two sample student T-test and values of P < 0.05 were considered as noteworthy and P < 0.005 as extremely noteworthy. All statistical techniques were executed using SPSS software version 20 [
Groups | Parameters | |||
---|---|---|---|---|
Cholesterol mg/dl | Triglycerides mg/dl | HDL-C mg/dl | LDL-C mg/dl | |
Control 10 ml/kg | 139.0 ± 1.30 | 249.8 ± 1.80 | 32.7 ± 0.68 | 179.8 ± 1.10 |
N. nucifera 50 mg/kg | 133.8 ± 0.40* | 223.1 ± 0.40** | 32.1 ± 0.74 | 170.3 ± 0.68** |
N. nucifera 100 mg/kg | 118.0 ± 0.65** | 180.5 ± 1.30** | 33.8 ± 0.34 | 152.0 ± 0.82** |
N. nucifera 200 mg/kg | 90.8 ± 0.86** | 142.0 ± 0.82** | 36.5 ± 1.1* | 129.3 ± 1.0** |
n = 7; Values are Mean ± S.E.M; *p < 0.05 noteworthy in comparison to control; **p < 0.005 extremely noteworthy in comparison to control.
Groups | Parameters | |||
---|---|---|---|---|
Cholesterol mg/dl | Triglycerides mg/dl | HDL-C mg/dl | LDL-C mg/dl | |
Control 10 ml/kg | 90.4 ± 1.0 | 187.8 ± 0.80 | 32.0 ± 0.72 | 144.4 ± 1.2 |
N. nucifera 50 mg/kg | 83.8 ± 0.74** | 172.0 ± 0.82** | 35.7 ± 1.2* | 133.1 ± 0.91** |
N. nucifera 100 mg/kg | 71.3 ± 0.89** | 151.4 ± 0.90** | 39.8 ± 0.40** | 112.4 ± 1.10** |
N. nucifera 200 mg/kg | 54.3 ± 0.97** | 131.0 ± 1.60** | 43.0 ± 0.53** | 92.4 ± 0.87** |
n = 7; Values are Mean ± S.E.M; *p < 0.05 noteworthy in comparison to control; **p < 0.005 extremely noteworthy in comparison to control.
No deaths of animal were recorded in any group all through the entire period of study. N. nucifera fruit ethanol extract at a dose of 50 mg/kg exhibited considerable lowering effects on TC after 30 days and extremely noteworthy effects after 45 days in comparison to control. The effects of the same dose of extract highly significantly lowered TG and LDL-C after 30 and 45 days whereas HDL-C was raised significantly after 45 days. N. nucifera fruit at a dose of 100 mg/kg revealed highly significant lowering effects on TC after 30 and 45 days as compared to control, whereas the TG and LDL-C lowering effects were also highly significant after 30 and 45 days in comparison to control. The HDL-C was highly significantly increased after 45 days but was not significantly altered after 30 days in comparison to control. N. nucifera fruit ethanol extract 200 mg/kg revealed highly noteworthy TC lowering effects after 30 and 45 days in comparison to control, whereas the TG and LDL-C lowering effects were also highly significant after 30 and 45 days in comparison to control. The HDL-C was significantly increased after 30 days and extremely considerably raised after 45 days as compared to control.
Hypercholesterolemia or high levels of cholesterol in the blood are primarily a metabolic derangement that can cause many diseases such as CVD and stroke. Hypercholesterolemia or hyperlipidemia may occur due to abnormalities in lipoproteins, the particles that bring cholesterol to the blood stream. Hyperlipidemia has a strong association with atherosclerosis [
Current study was distinctively planned to assess the hypolipidemic activities of N. nucifera fruit after 30 and 45 days in rabbits kept on high cholesterol diet for 30 days. The results showed remarkable cholesterol lowering effects of N. nucifera fruit extract at doses 50, 100 and 200 mg/kg but were particularly dominant at 200 mg/kg at which fruit extract highly significantly reduced TC, TG and LDL-C after 30 and 45 days and increased HDL-C significantly after 30 days and highly significantly after 45 days.
Previous studies have established an inverse relationship between flavonoids intake and total lipid concentration in plasma. Diet rich in flavonoids protects against coronary artery diseases [
Previous studies on anti-obesity and hypolipidemic activities of N. nucifera seed resulted in inhibition of lipid accumulation and reduced expression of peroxisome proliferator activated receptor gamma (PPAR), leptin in cultured human adipocytes and glucose transporter (GLUT 4) indicating the inhibitory effect on the differentiation of pre-adipocytes in to adipocytes [
N. nucifera fruit has exhibited remarkable anti-hyperlipidemic activity which may be due to the presence of secondary metabolites especially flavonoids, saponins, tannins and procyanidin in both segments i.e. seeds and pods of this fruit which may be useful in managing conditions such as CVDs and stroke but more preclinical and clinical studies should be encouraged to confirm these findings.
The authors are thankful to Dr. Mohtesheem, Associate Professor and Chairman Department of Pharmacognosy for identification and authentication of N. nucifera fruits. The authors are also thankful to Prof. Dr. Iqbal Chaudary, Director, HEJ, Research Institute of Chemistry for granting permission to use Institute laboratory and workshop facilities.
The authors declare no conflict of interest.
Rajput, M.A., Khan, R.A. and Assad, T. (2017) Evaluation of the Anti-Hyperlipidemic Activity of Nelumbo nucifera Fruit in Rabbits Fed with High Cholesterol Diet. Pharmacology & Pharmacy, 8, 205-213. https://doi.org/10.4236/pp.2017.86014