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 »  Abstract
 » Introduction
 »  Materials and Me...
 » Results
 » Discussion
 » Conclusion
 »  References
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 Table of Contents    
SHORT COMMUNICATION
Year : 2022  |  Volume : 54  |  Issue : 2  |  Page : 126-130
 

Pharmacological evaluation of embelin - chitosan nanoparticles as an antidiabetic agent


1 Associate Professor, Sri Ramachandra Faculty of Pharmacy, Sri Ramachandra Institute of Higher Education and Research (Deemed University), Porur, Tamil Nadu, India
2 Professor and Head, Department of Biochemistry, St. Peter's Institute of Higher Education and Research, Avadi, Tamil Nadu, India
3 Professor, Sri Ramachandra Faculty of Pharmacy, Sri Ramachandra Institute of Higher Education and Research (Deemed University), Porur, Tamil Nadu, India
4 Sr. Principal Scientist, Head & Professor – AcSIR, Microbiology Division, Central Leather Research Institute, Adyar, Chennai, Tamil Nadu, India

Date of Submission05-Feb-2020
Date of Decision11-Jun-2021
Date of Acceptance24-Mar-2022
Date of Web Publication10-May-2022

Correspondence Address:
Dr. Arumugam Gnanamani
Microbiology Division, Central Leather Research Institute, Adyar, Chennai - 600 020, Tamil Nadu
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/ijp.ijp_47_20

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 » Abstract 


Embelin has been reported to possess variety of pharmacological activities such as androgenic antagonists, antiangiogenic, antibacterial, anticancer, anticonvulsant, antidiabetic, antidepressant, antihelmintic, antifertility, antihyperlipidemic, anti-inflammatory, antimalarial, antimitotic, antiobesity and antioxidant properties. The current research work aimed to study the hypoglycemic effect of embelin-chitosan nanoparticles (ECNPs) diabetic rats provoked by streptozotocin (STZ). Embelin nanoparticles (ENPs) were created by combining chitosan, a natural biopolymer, and glutaric acid, a new cross-linker. STZ 50 mg/kg was given intravenously into Sprague-Dawley rats weighing 250–300 g (75–90 days) to induce experimental diabetes. The antidiabetic efficacy of orally administered ECNPs in diabetic rats developed by STZ was investigated, as well as histological examination. When compared to diabetic control rats, ECNPs (25 mg/kg body weight and 50 mg kg body weight) and standard glibenclamide (10 mg/kg body weight) treated rodents exhibited a remarkable drop in glucose contents. Furthermore, histological research showed that ECNPs-treated rats were harmless up to amount of 25 mg/kg bwt. Thus current investigation reveals that ECNPs have antidiabetic potential and may be beneficial in treating hyperglycemia in people.


Keywords: Antidiabetic, chitosan, embelin, histopathological, nanoparticles, plasma glucose


How to cite this article:
Maanvizhi S, Radhakrishnan N, Krishnan C, Gnanamani A. Pharmacological evaluation of embelin - chitosan nanoparticles as an antidiabetic agent. Indian J Pharmacol 2022;54:126-30

How to cite this URL:
Maanvizhi S, Radhakrishnan N, Krishnan C, Gnanamani A. Pharmacological evaluation of embelin - chitosan nanoparticles as an antidiabetic agent. Indian J Pharmacol [serial online] 2022 [cited 2022 May 23];54:126-30. Available from: https://www.ijp-online.com/text.asp?2022/54/2/126/344962





 » Introduction Top


Diabetes mellitus, also known as diabetes, is a metabolic illness that produces high amounts of blood sugar. It is categorized into Type I (due to-cell destruction, generally resulting in absolute insulin shortage) and Type II (due to-cell destruction, usually resulting in absolute insulin deficit) (because of a growing insulin secretory malfunction in the context of insulin resistance).[1] By the end of 2030, the global and regional diabetes population is predicted to grow at an alarming rate of 10.2% (578 million).[2] Diabetes Type 2 is a chronic metabolic condition that is rapidly becoming a primary cause of morbidity and mortality. The majority of the existing oral dose forms for diabetes have various and sometimes life-threatening side effects; hence, treating diabetes without these side effects is a significant issue.[3]

As a result, numerous researchers are working in traditional and complementary medicine to cure diabetes. Interestingly, 400 traditional plants have shown to possess antidiabetic activity so far.[4] However, not all herbal treatments have scientific evidence to support their usefulness in treating diabetes.[5] Embelia ribes is a well-known and thoroughly explored herbal plant for a variety of pharmacological actions. Embelin is a natural benzoquinone (2, 5-dihydroxy-3-undecyl-p-benzoquinone) that was discovered to be a major ingredient of Embelia ribes and has been linked to a variety of pharmacological properties.[6],[7] Embelin has been demonstrated to be nontoxic and capable of rapidly reducing and stabilizing metal ions to nanoparticles. These embelin-stabilized nanoparticles were found to be biocompatible with human blood and cells.[8] The purpose of this study is to see how embelin-chitosan nanoparticles (ECNPs) affect plasma glucose levels and histopathological alterations in streptozotocin (STZ)-induced diabetic mice.


 » Materials and Methods Top


Animal groupings/experimental animals

For the current in vivo animal investigation, 150–200 g of female Sprague-Dawley rats were obtained from the SRMC central animal house in Chennai and maintained in regular laboratory settings (temperature 24°C ± 2° C; humidity 45 ± 10%) with a 12 h day and night cycle, respectively. All experimental rats were fed with standard laboratory diet and had unrestricted access to drinking water. Then, assessment of antidiabetic activity was carried out with the agreement of the (SRM-IEC) and in consent with the institutional ethical instruction for the safeguard of animals at SRMC, Chennai (IAEC/XXXVII/SRU/342/2014).

Isolation of embelin

Embelin was extracted from Embelia ribes berries and described using Radhakrishnan's techniques (2012).[9]

Chemicals

Chemicals such as chitosan, glibenclamide, glutaric acid, sodium azide, sodium carboxy methylcellulose (CMC), and streptozotocin (STZ) were procured from Sigma-Aldrich, USA. Similarly biochemical reagents for determination of plasma glucose, creatinine, and urea were procured from Radiant Specialty Diagnostics (Accurex Biomedical Reagents), Nungambakkam, Chennai.

Preparation of embelin-chitosan nanoparticles

ECNPs were synthesized and characterized in accordance with previous reports.[10],[11]

In vivo experimental design

The rats were sorted into five batches of 6 (female) rats each group. Group I rats were healthy and served as standard controls, with 0.5% CMC functioning as a vehicle. Rats from Groups II, III, IV, and V were given a single dose intraperitoneal injection of STZ (STZ - 50 mg/kg body weight) and served as untreated diabetes controls (0.5% CMC), diabetic glibenclamide, diabetic ECNPs (minimum dose - 25 mg/kg/day), and diabetic ECNPs (maximum dose - 50 mg/kg/day). The experiment was conducted 2 days after STZ injection in diabetic rats with glycosuria and hyperglycemia (blood glucose values of 200–300 mg/ml). The rats were starved overnight before blood was drawn from the tail vein. Groups III, IV, and V received glibenclamide (10 mg/kg/day), ECNPs (25 mg/kg/day), and ECNPs (50 mg/kg/day) orally. After 2 weeks, the rats were slaughtered and blood samples were drawn from the heart. Plasma glucose levels were measured at 0, 7, and 14 days after an overnight fast. On the 14th day, the 24 h urine was collected by placing rats in individual metabolic cages and utilizing sodium azide as a preservative. Following drug administration, the levels of plasma and urine glucose, creatinine, and urea were measured in urine.

Histopathological examination

Following the experimental period, the collected organs (pancreas, liver, and kidney) were fixed in 10% buffered neutral formalin solution for 48 h before being processed using the standard inclusion into paraffin technique. The preparations were then stained with hematoxylin-eosin (H and E) and histopathologically evaluated under a microscope, while the remaining sections were frozen at 80°C.

Biochemical analysis

Accurex kits (Radiant Specialty Diagnostics, Nungambakkam, Chennai) were used to estimate blood glucose levels as well as biochemical parameters in urine such as glucose, creatinine, and urea levels.

Statistical analysis

A one-way variance analysis was performed using SPSS version 3.5 (IBM SPSS Statistics., Chicago, USA). For group comparisons, a post hoc Tukey's test was used. With P = 0.001, values were considered statistically significant.


 » Results Top


Extraction and chemical characterization of embelin (active principle of Embelia ribes)

Embelin was extracted from Embelia ribes berries and chemically analyzed utilizing a range of tools in this study (thin-layer chromatography, ultraviolet, Fourier transform infrared, differential scanning calorimetry, thermogravimetry, and nuclear magnetic resonance). The current study, however, did not demonstrate the findings.

Physicochemical characterization of NPS

Embelin nanoparticles (ENPs) were created in this study using the natural biopolymer chitosan and the new cross-linker glutaric acid. The prepared ECNPs were discovered to be of a discrete, free-flowing nature. As shown in [Table 1], approximately three batches of ECNPs were prepared. The percentage yields of ECNPs ranged from 75.23 to 78.45 ± 1.0106 % [Table 1], while the encapsulation efficiency of ECNPs ranged from 71.64 to 75.50 ± 1.1908 % [Table 1].
Table 1: Composition, yield, and entrapment efficiency of embelin loaded chitosan nanoparticles

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The relevance of the in vitro drug release evaluation results has shown the product quality and performance of the offered drug.

Embelin release investigations in the current study show that the release was uniform, and the amount of drug released at the end of 6 h was judged to be acceptable and sustainable [Table 2] and [Figure 1]. Further, transmission electron microscopy analysis shows that a well dispensability of ECNPs pointing outward with size up to 100 nm was observed as shown in [Figure 2].
Table 2: In vitro release profiles of embelin loaded chitosan nanoparticles

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Figure 1: Cumulative percentage release of embelin nanoparticles

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Figure 2: TEM image of embelin nanoparticles

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Antidiabetic activity screening in normal and streptozotocin-induced diabetic rats

STZ-induced diabetic rats were used to test the antidiabetic activity of ECNPs. The data obtained on the fasting blood sugar level of normal and diabetic rats of antidiabetic effects of ECNPs are depicted in [Table 3] and [Table 4]. Moreover, in our study, there was a significant reduction of plasma and urine glucose level was observed in all the treatment groups compared to standard [Table 3] and [Table 4]. There is no statistically significant different among the groups at the 0th day and 7th day except standard treated rats.
Table 3: Estimation of plasma glucose level

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Table 4: Estimation of urine glucose, urea, and creatinine level on 14th day

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However, a statistically significant reduction in the measurement of plasma glucose of the standard and treatment groups compared to Group II was revealed at the 14th day (diabetic control animals). Glibenclamide demonstrated a slower onset of hypoglycemic action and showed a sustained decrease in blood sugar levels. ECNPs at doses of 25 and 50 mg/kg body weight decreased blood sugar levels significantly and were found to be more effective. The treatment groups exhibited dose-dependent effects. Diabetes, as depicted/demonstrated in [Table 3], resulted in a significant initial increase in fasting blood glucose levels across the board.

Histopathological studies

For this purpose the obtained liver, kidney and pancreas were fixed in 10% buffered neutral formalin solution for 48 hours and then included in paraffin by the usual technique. It was processed by the usual technique of inclusion into paraffin. The preparations were then stained with Haematoxylin-Eosin and they were examined under microscope for histopathological evaluation [Figure 3], [Figure 4], [Figure 5], whereas the remaining sections were frozen at −80°C.
Figure 3: Histopathological change in kidney of embelin nanoparticles on Sprague-Dawley rats

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Figure 4: Histopathological change in liver of embelin nanoparticles on Sprague-Dawley rats

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Figure 5: Histopathological change in pancreas of embelin nanoparticles on Sprague-Dawley rats

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 » Discussion Top


Embelin from Embelia ribes has numerous applications in Indian medicine. ENPs were created in this study using the natural biopolymer chitosan and the new cross-linker glutaric acid. Similarly, in our prior investigation, we reported ECNPs that were cross-linked with glutaraldehyde.[11] At the end of the 6th h, the in vitro cumulative percent drug release was reported to be 61.43%. As a result, the current study's findings were consistent with previous reports.[11] The doctoral thesis report includes a chemical characteristic study of manufactured ECNPs.[12] Embelin has been shown to have antidiabetic properties in both alloxan and STZ-induced diabetic rats. STZ-induced hyperglycemia, on the other hand, has been regarded as a helpful experimental paradigm for studying the activity of hypoglycemic medications.[12] Similarly, embelin has been shown in silico to decrease human pancreatic alpha-amylase and human aldose reductase activity.[13],[14] According to the findings, ECNPs have strong antidiabetic effect in both normal and STZ-induced hyperglycemic rats. Diabetes mellitus was produced by STZ injection, most likely due to elimination of the beta cells of the pancreatic  Islets of Langerhans More Details. Oral administration (per os) of the ECNPs at doses of 25 and 50 mg/kg body weight exhibited no remarkable change in behavior, showing that the ECNPs are not harmful in tested animals at the observable conditions. Oral delivery of ECNPs resulted in no toxicity, mortality, or behavioral abnormalities for up to 14 days.[15] The standard showed the greatest reduction, followed by ECNPs (25 mg/kg), hence the current study results were consistent with previous publications.[15],[16] Similarly, the current study reveals that ECNPs have antidiabetic potential and may be beneficial in treating hyperglycemia in people. Both serum creatinine and urea levels were excessively high, indicating compromised kidney function.[17] Histopathological examination also reveals that ECNPs are safe at a dose of 25 mg/kg body weight. The current study's findings were consistent with previous publications.[17] Long-term research of ECNPs is thus required to establish the specific mechanism of action to develop it as a commercially accessible powerful antidiabetic medicine to protect body organs from diabetic alterations.


 » Conclusion Top


Embelin from Embelia ribes has numerous applications in Indian medicine. Oral administration (per os) of the ECNPs at doses of 25 and 50 mg/kg body weight results in no remarkable changes in behavior, showing that the ECNPs are not hazardous in tested animals at the observable conditions. In conclusion, the findings indicated that ECNPs had antidiabetic potential and may be beneficial in treating hyperglycemia in people.

Acknowledgment

This research work was partially supported by a Young Faculty Research Grant (C. No. 36/Dean/2012) provided by the Sri Ramachandra University, Porur, Chennai. The authors would like to thank SRU management and also CLRI for helping us to carry out this work.

Financial support and sponsorship

This research work was partially supported by a Young Faculty Research Grant (C. No. 36/Dean/2012) provided by the Sri Ramachandra University, Porur, Chennai.

Conflicts of interest

There are no conflicts of interest.



 
 » References Top

1.
Kalekar SA, Munshi RP, Bhalerao SS, Thatte UM. Insulin sensitizing effect of 3 Indian medicinal plants: An in vitro study. Indian J Pharmacol 2013;45:30-3.  Back to cited text no. 1
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Balekari U, Veeresham C. Insulinotropic agents from medicinal plants. J Pharm Sci Emerg Drugs 2013;2:2-1.  Back to cited text no. 2
    
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Jung M, Park M, Lee HC, Kang YH, Kang ES, Kim SK. Antidiabetic agents from medicinal plants. Curr Med Chem 2006;13:1203-18.  Back to cited text no. 3
    
4.
Bailey CJ, Day C. Traditional plant medicines as treatments for diabetes. Diabetes Care 1989;12:553-64.  Back to cited text no. 4
    
5.
Morelli V, Zoorob RJ. Alternative therapies: Part I. Depression, diabetes, obesity. Am Fam Physician 2000;62:1051-60.  Back to cited text no. 5
    
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Radhakrishnan N, Gnanamani A, Mandal AB. A potential antibacterial agent, Embelin – A natural benzoquinone extracted from Embelia ribes. Biol Med 2011;3:1-7.  Back to cited text no. 6
    
7.
Radhakrishnan N, Kavitha V, Raja ST, Gnanamani A, Mandal AB. Embelin – A natural potential cosmetic agent. J Appl Cosmetol 2011;29:99-107.  Back to cited text no. 7
    
8.
Sasidharan S, Poojari R, Bahadur D, Srivastava R. Embelin-mediated green synthesis of quasi-spherical and star-shaped plasmonic nanostructures for antibacterial activity, photothermal therapy, and computed tomographic imaging. ACS Sustain Chem Eng 2018;6:10562-77.  Back to cited text no. 8
    
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Radhakrishnan N, Gnanamani A, Prasad NR, Mandal AB. Inhibition of UVB-induced oxidative damage and apoptotic biochemical changes in human lymphocytes by 2,5-dihydroxy-3-undecyl-1,4-benzoquinone (embelin). Int J Radiat Biol 2012;88:575-82.  Back to cited text no. 9
    
10.
Arumugam G, Krishnan C, Babu M, Maanvizhi S. Preparation and Characterization of Embelin Nanoparticle. 2nd International Conference on Chemical, Environmental and Biological Sciences (ICCEBS'2013) March 17-18, 2013 Dubai (UAE); 2013.  Back to cited text no. 10
    
11.
Maanvizhi S. Preparation, Characterization and Antidiabetic Efficacy of Nanoparticles of Natural Benzoquinone and Its Metal Complexes an Approach Under in vitro Docking and in vivo Animal Model Studies (Ph.D thesis submitted to Sri Ramachandra University, Chennai); 2015.  Back to cited text no. 11
    
12.
Szkudelski T. The mechanism of alloxan and streptozotocin action in B cells of the rat pancreas. Physiol Res 2001;50:537-46.  Back to cited text no. 12
    
13.
Maanvizhi S, Narayanaswamy R, Wai LK, Gnanamani A. HPAA inhibitory effect of embelin and its metal complexes on diabetic complications: An approach with molecular docking studies. J Chem Pharm Res 2014;6:679-82.  Back to cited text no. 13
    
14.
Maanvizhi S, Narayanaswamy R, Wai LK, Gnanamani A. Molecular docking analysis of embelin and its metal complexes as human aldose reductase (HAR) inhibitor. Pharm Lett 2014;6:165-8.  Back to cited text no. 14
    
15.
Gupta R, Sharma AK, Sharma MC, Dobhal MP, Gupta RS. Evaluation of antidiabetic and antioxidant potential of lupeol in experimental hyperglycaemia. Nat Prod Res 2012;26:1125-9.  Back to cited text no. 15
    
16.
Naik SR, Niture NT, Ansari AA, Shah PD. Anti-diabetic activity of embelin: Involvement of cellular inflammatory mediators, oxidative stress and other biomarkers. Phytomedicine 2013;20:797-804.  Back to cited text no. 16
    
17.
Ronco C, Grammaticopoulos S, Rosner M, De Cal M, Soni S, Lentini P, et al. Oliguria, creatinine and other biomarkers of acute kidney injury. Contrib Nephrol 2010;164:118-27.  Back to cited text no. 17
    


    Figures

  [Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5]
 
 
    Tables

  [Table 1], [Table 2], [Table 3], [Table 4]



 

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