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Phytotherapeutic Potential of Bi-Herbal Extract of Cinnamon and Turmeric: in Vivo Antidiabetic Studies R

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Phytotherapeutic Potential of Bi-Herbal Extract of Cinnamon and Turmeric: in Vivo Antidiabetic Studies R Sivaranjani et al. Clinical Phytoscience (2021) 7:38 https://doi.org/10.1186/s40816-021-00275-3 ORIGINAL CONTRIBUTION Open Access Phytotherapeutic potential of bi-herbal extract of cinnamon and turmeric: in vivo antidiabetic studies R. Sivaranjani*, T. John Zachariah and N. K. Leela Abstract Background: The spices, cinnamon and turmeric have both culinary and pharmaceutical applications. Earlier studies proved their effect on reducing the symptoms associated with type 2 diabetes, a major lifestyle disease affecting millions world over. In our work, we prepared a bi-herbal extract of cinnamon and turmeric and studied its effect to alleviate the symptoms of type 2 diabetes in rat model. Method: The phenolic rich bi-herbal extract was given to diabetes induced male wistar rats for 28 days at two different concentrations (50 and 150 mg/Kg bwt.). The changes in blood glucose level were monitored at weekly interval. At the end of the experiment, blood serum was collected and used for the estimation of lipid profile, Glucose, aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), Total Protein (TP), Albumin, Urea and Creatinine levels. Result: The serum analysis revealed that administration of the bi-herbal extract at 150 mg/Kg bwt. significantly reduced the blood glucose level (152.60 ± 40.2 mg/dL; p < 0.05) of diabetic animals as compared to diabetic control (335.40 ± 67.3 mg/dL; p < 0.05) at the end of 4 weeks. The treatment also resulted in the reduction of total cholesterol and LDL-cholestrol levels as compared to diabetic control animals. The histopathological examination of tissues showed the improvement in pancreatic architecture and restoration of tissue integrity in liver and kidney. Conclusion: The study concludes that methanol extract of combination of cinnamon and turmeric has good hypoglycemic, hypolipidemic and organ protective potential in diabetic rats which could be developed into functional food supplementation for the prevention of type 2 diabetes. Keywords: Cinnamon, Turmeric, Streptozotocin, Antidiabetic, Wistar rats Background insulin injection throughout their life, whereas incidence Non-Insulin Dependent Diabetes Mellitus (NIDDM) of NIDDM can be reduced through changes in food popularly known as Type-2 diabetes is a biochemical habits and lifestyle [3]. Since NIDDM is a complex metabolic disorder wherein the ability of cells to utilize metabolic disorder, single drug treatment options are blood glucose is reduced leads to metabolic abnormal- found to be not effective; moreover prolonged use of ities [1]. By 2025, around 300 million people were esti- synthetic drugs could lead to adverse health impacts. mated to be affected by diabetes mellitus [2]. People Hence, the search for new phytotherapeutic agents from with Insulin-Dependent Diabetes Mellitus (IDDM), natural sources in the form of bioactive compounds has otherwise known as Type-1 diabetes have to rely on assumed global significance [4]. In this regard, herbs and spices are emerged as panacea for many ailments occur- ring due to lifestyle modifications. * Correspondence: [email protected] True Cinnamon, scientifically known as Cinnamomum Division of Crop Production and Post-Harvest Technology, ICAR – Indian Institute of Spices Research, Kozhikode, Kerala 673012, India zeylenium or Cinnamomum verum is reported to have © The Author(s). 2021 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. Sivaranjani et al. Clinical Phytoscience (2021) 7:38 Page 2 of 9 numerous biological activities including antifungal, anti- extract (hereafter mentioned as CTE) has the highest microbial, antioxidant, anticancerous, antiviral, antidia- total phenolic content and showed highest antioxidant betic and immunomodulatory activities, to name a few activity (previously published work from our laboratory) [5–8] validated through both in vitro and in vivo assays. [22], were taken for the present study. The suspension Few clinical studies in humans also proved its effect of the extract was prepared by dissolving it in 5% against type 2 diabetes [9, 10]. Hayward et al. [11] com- DMSO, a universally accepted vehicle in oral feeding pared the anti-hyperglycaemic properties of the four studies. major commercial cinnamon types used around the world namely Cinnamomum cassia (Chinese cinnamon), HPLC quantification of curcuminoids and cinnamic acid in C. burmanii (Indonesian cinnamon), C. loureirii CTE (Vietnamese cinnamon) and C. zeylanicum (Ceylon cin- CTE was dissolved completely in HPLC grade methanol namon). Among them, C. zeylanicum showed the great- in the concentration of 1 mg/mL and filtered using est potential to inhibit the formation of advanced 0.45 μm filter before injecting into HPLC. The separ- glycation end products (AGEs) during digestion thus ation of curcuminoids was achieved in Purospher™ RP proved its antihyperglycemic activity. Cinnamon con- C18 column with the dimension of 150 × 4.6 mm, 5 μm tains numerous bioactive compounds of phenolics and (Merck, Germany) using a shimadzu LC-20 HPLC sys- flavonoids category which individually or synergistically tem (Shimadzu, Kyoto, Japan) consists of a binary pump- found to have antidiabetic properties [8]. ing system and photodiode array (PDA) detector. The Like cinnamon, the golden spice turmeric is also re- mobile phase used was Acetonitrile and 0.1% orthophos- ported to have multifaceted biological activities includ- phoric acid (50:50) and eluted at a flow rate of 1 mL/min ing antidiabetic potential [12]. The mode of action of in an isocratic mode. The resulting chromatogram data curcumin, the phytocompound of interest from turmeric was processed using shimaduz LC solution 3.6 software. against diabetic signalling pathway was well-worked out Elution of the compounds was monitored at 425 nm for in the past decade [13–16]. curcuminoids and 274 nm for cinnamic acid and con- Few reports are available on the synergistic activity of centration of these compounds were calculated by com- plant based phenolics among themselves and also with paring the peak area of the standards. The CTE did not other oral antidiabetic drugs for diabetes treatment [17– contain cinnamaldehyde. We hypothesized that the 21]. Yet, it is quite difficult to pinpoint the cause and ef- rigorous process involved in the extract preparation fect relationship of individual phenolic compounds in led to the loss of cinnamaldehyde in it. the extract of combined spices on any disease models. The question of which spices to select for combination Experimental animals poses barriers which could be overcome by careful lit- The experiment was undertaken at animal house facility erature survey and associated traditional knowledge on of CARe – Keralam Ltd., Thrissur, Kerala following use of those spices. Though individual extract of tur- guidelines of Institute Animal Ethics Committee (CKL/ meric and cinnamon in reducing the effect of diabetes in TOX/IAEC/2017–3/95). Thirty albino male wistar rats experimental animals have established with numerous were utilized in this study. Animals were maintained work, the combination of these two spices to that effect under standard laboratory conditions (22 ± 3 °C room has not been tried so far. With this background, the temperature and 50–60% humidity) with alternating present study was undertaken to study the effect of light and dark cycles of 12 h. The rats were fed with pel- phenolic rich bi-herbal extract of cinnamon and tur- let diet ad libitum and were acclimatized to laboratory meric on STZ induced diabetes in wistar rats. conditions for 7 days prior to the experiment. Induction of diabetes was done using single intraperitoneal injec- Materials and methods tion of 30 mg/kg bwt. of streptozotocin (STZ) dissolved Plant collection and preparation in freshly prepared citrate buffer (0.1 M, pH 4.5). Ani- Turmeric variety, IISR Prathibha with high curcumin mals with fasting blood glucose over 200 mg/dL, 3 days content (~ 5%) released from ICAR - IISR and dried bark after STZ administration were considered diabetic and of true cinnamon, Cinnamomum zeylenicum collected they were divided into five groups of 6 rats each for CTE from Alakkode (Kannur) estate of Plantation Corpor- treatment. ation of Kerala Limited were used in the study. The sam- ples were identified and authenticated by the Institute’s Treatment details taxonomist. The dried samples of cinnamon and tur- meric were coarsely ground and mixed in 2:1 ratio and Group 1: Normal control were used in the preparation of sequential extracts Group II: Diabetic control through soxhlet apparatus. Among them, methanol Group III: CTE I (50 mg/kg bwt.) Sivaranjani et al. Clinical Phytoscience (2021) 7:38 Page 3 of 9 Group IV: CTE II (150 mg/kg bwt.) sacrificed at scheduled termination. Histopathological Group V: Glibenclamide positive control (5 mg/kg analysis of the stained tissue samples was done using bwt.) light microscope. The acute oral toxicity study of the extract in female wistar rats at 2000 mg/kg bwt. did not reveal any clinical Statistical analysis signs, mortalities and gross pathological changes due to All the results were expressed as mean ± SEM. The data toxicity (data not shown). Since the IAEC suggested analysis were performed using statistical software pack- using minimum animals, one lower dose and one higher age SPSS version 16 to determine the significant differ- dose was taken based on acute oral toxicity study.
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