INTRODUCTION:

Obesity is brought about by an awkwardness between vitality admission and utilization. It is as often as possible related with dyslipidemia, cardiovascular dangers, hypertension, and type-2 diabetes mellitus, and in this way is perceived as one of the most genuine general wellbeing problems¹. Adiponectin is an adipocyte-inferred hormone². Late genome-wide outputs have mapped a vulnerability locus for sort 2 diabetes and metabolic disorder to chromosome 3q27, where the quality encoding adiponectin is found. Here we demonstrate that diminished articulation of adiponectin associates with insulin opposition in mouse models of modified insulin sensitivity³.

Adiponectin diminishes insulin obstruction by diminishing triglyceride content in muscle and liver in hefty mice^4,5 This impact results from the expanded articulation of particles associated with both unsaturated fat ignition and vitality scattering in muscle. In addition, insulin opposition in lipoatrophic mice was totally turned around by the blend of physiological dosages of adiponectin and leptin, however just somewhat by either adiponectin or leptin alone⁶. We presume that diminished adiponectin is ensnared in the advancement of insulin opposition in mouse models of both corpulence and lipoatrophy. This information likewise demonstrates that the renewal of adiponectin may give a novel treatment methodology to insulin opposition and type 2 diabetes. Various medications focused towards restraint of amylase/α-glucosidase/lipase, loss of hunger, and improving unsaturated fat digestion, have been affirmed for the treatment of corpulence; be that as it may, the greater part of them have been pulled back from the market on account of their genuine unfriendly effects⁷. Along these lines, numerous investigations have been directed to discover and build up another enemy of stoutness sedate or a dietary enhancement with lesser side effects^8,9.

The plant Syzygium cerasoideum has wide medicinal uses as anti-rheumatic, hypoglycemic, rubefacient, back- unpleasant astringent is given in diarrhea and bronchitis. A concentrate of the root infusion is applied and rubbed over difficult joints. Aerial parts show hypoglycemic action. Syzygium cerasoideum comprises of phenolic mixes cyanidin, 3-glucoside, delphinidin 3-glucoside, ellagic corrosive, kaempferol, myricetin, quercetin and rutin¹⁰. Based on the above findings, Syzygium cerasoideum extracts was evaluated for it’s antiobesity/ antidiabetic effects on experimental induced obesity.

MATERIALS AND METHODS:

Preparation of S. cerasoideum extracts:

The plant was authenticated by Dr Madhava Chetty, SVU Tirupathi was voucher specimen number 1320 was deposited for future reference and are prepared for successive extraction with Pet-ether (40-60⁰ C), Chloroform, Ethyl acetate, and Methanol. The concentrates were set up by nonstop extraction utilizing soxhlet unit. The four concentrates were then thought under diminished strain to evacuate the dissolvable totally and lyophilized to be free from solvent and water¹¹. Based on the earlier study the chloroform and methanol extracts were used in the study¹²

Phytochemical analysis:

Phytochemical analysis was performed for all the extracts which include a test for Steroids, Triterpenoids, Glycosides, Flavonoids, Saponins, Alkaloids, Tannins, Carbohydrates, Fat and oil according to the standard protocol¹³.

Experimental procedure:

Eight week old male C57BL/6J animals were procured from Hylasco, Hyderabad, India. Thirty Wistar rats 100-150g were obtained from the Animal House, Dayananda Sagar University, Bengaluru. Animals were housed in polypropylene cages with provision for water bottle holder and feed hopper with corn cobs as bedding material, under standard air-conditioned laboratory conditions, Temperature of 24 ºC ± 1ºC, Relative humidity of 63% to 48%,12 h light/12 h dark cycle. The maximum and minimum temperature and relative humidity in the experimental room were recorded once daily. libitum Pelleted rodent feed (Teklad global, 14% protein, maintenance feed) and HFD. Water from Deep bore well water passed through charcoal filters and exposed to UV rays and water in polypropylene water bottles were provided to the animals.

Every single trial convention including creatures were endorsed by IAEC DSU/Ph.D./IAEC/13/2017-18 and all tests were led according to standards and rules of the CPCSEA, India.

In the present investigation, an eating routine incited stoutness mouse model was created by bolstering the HFD to C57BL/6J mice. The improvement of the hyperglycemic state was affirmed by checking the fasting blood glucose at one-week normal interims.

Study Design:

a) Oral Glucose Tolerance Test:

The key point is that more diabetics are beginning from a populace with typical fasting blood glucose than from subjects with disabled fasting glucose, the individuals who can be recognized by the new ADA proposals. Then again, the OGTT identifies all the more effectively early diabetes just as subjects with IGT, as the glycemia at the subsequent hour appears to be significant as a symptomatic instrument. All preliminaries went for sort 2 diabetes counteractive action included IGT subjects, who couldn't be perceived without OGTT. It appears to be in this way obvious that the normal execution of OGTT is by and by the main conceivable answer. The oral glucose resilience test has regularly been utilized to assess obvious insulin discharge and insulin obstruction in different clinical settings. Be that as it may, on the grounds that insulin affectability and insulin discharge are related. The most dependable information on the predominance of diabetes depend on two-hour plasma glucose esteems after an oral glucose resilience test, which is at present the highest quality level epidemiological and clinical symptomatic test for diabetes and disabled glucose tolerance¹⁴.

Oral glucose resistance test was trailed by the strategy for Prakasam et al¹⁵. (2003)

Animals were divided into 5 groups with 6 animals per group.

Group I : Normal control (just vehicle)

Group II : Received 200mg/kg MESC orally \

Group III : Received 400mg/kg MESC orally

Group IV : Received 200mg/kg CESC orally

Group V : Received 400mg/kg CESC orally

Every one of the animals were given oral organization of glucose (2g/kg) 60min after dosing. Blood tests were gathered from the retro-orbital plexus just before 0h and at 30min, 60min, 90min, and 120min after the glucose stacking, and blood glucose levels were evaluated. The values were communicated as mean ± SEM, and a diagram of time in minutes versus glucose focus was plotted.

b) Screening of concentrates for Antidiabetic/Anti-Obesity in C57BL/6 J mice

C57BL/6J mice:

Type 2 diabetic model by just sustaining high-fat feed to non-hefty, non-diabetic C57BL/6J mouse strain was at first created in Japan and is presently accessible at Jackson Laboratory, Bar Harbor. It is portrayed by checked corpulence, hyperinsulinaemia, insulin obstruction and glucose intolerance¹⁶. Moreover, they show stamped fasting just as basal hyperglycaemia rather than typical basal glucose level seen in C57BL/6J (ob/ob) mice. These mice are shown to create fringe leptin opposition. They show the vast majority of the trademark highlights of the patients with a hereditary inclination to create type 2 diabetes when they become large. This creature model speaks to both hereditary and ecological hazard factors rather than C57BL/6J (ob/ob) mouse in which the beginning of side effects is exceptionally hereditarily determined¹⁷. Further, its handiness for medication testing has been accounted for in the writing as these mice treated with an orally dynamic inhibitor of dipeptidyl peptidase-IV (LAF237) are appeared to have standardized glucose resistance in relationship with expanded insulin secretion¹⁸.

Following a high-fat eating regimen for 30days, the animals were randomized into explicit treatment groups.

Animals were partitioned into 6 groups with 6 animals for each group.

Group I : Lean control (typical chow diet)

Group II : HFD control

Group III : Received 200mg/kg MESC orally

Group IV : Received 400mg/kg MESC orally

Group V : Received 200mg/kg CESC orally

Group VI : Received 400mg/kg CESC orally

The animals in the lean control group were bolstered typical chow diet, while animals in the HFD control and treatment gatherings were encouraged with HFD all through the exploratory period, and all treatments were allowed once per day orally for 30days.

Assessment parameters

Bodyweight

The body weight was recorded once week by week, from the day of study commencement to the end.

Blood glucose estimation

All creatures have fasted for 6hr, and fasting blood glucose levels were estimated utilizing Accu-check glucometer (Roche Diagnostics) consistently until the part of the arrangement.

Estimation of Triglycerides:

Following a month of treatment, all animals have fasted for 6hr, and under isoflurane anesthesia, blood tests were gathered by retro-orbital cut and permitted to cluster for 30min at room temperature, centrifuged at 10000rpm for 10min at 4º C, and the serum was gathered for investigation.

The glycerol and free unsaturated fats were assessed by colourimetric technique utilizing monetarily accessible free unsaturated fat investigation unit (Sigma-Aldrich, Banglore) and Randox pack separately.

RESULTS AND ANALYSIS:

a. Oral Glucose Tolerance Test

Table 1: The effects of Syzygium cerasoideum extracts on Oral Glucose Tolerance Test

Groups	Dose	Glucose concentration in mg/dL
Groups	Dose	0 hr	0.5 hr	1hr	1.5 hr	2 hr
Group I	Control	100±1.81	84±0.7	102±1.6	94±1.02	93±0.91
Group II	MESC 200mg/kg	100±2.04	89±2.76	97±3.70	102±5.03	94±1.53
Group III	MESC 400mg/kg	100±3.62	93±3.82^*	94±1.14	100±3.51	95±1.02
Group IV	CESC 200mg/kg	98±3.04	84±1.06	96±1.90	94±0.57	92±0.76
Group V	CESC 400mg/kg	99±3.84	88±1.83	94±1.16	97±2.56	96±1.68

^*p˂0.05 when compared with the control

Values are expressed as Mean± SEM. All the data were statistically treated with Dunnet Multiple comparison test post one way ANOVA using graph pad version 5.0

Figure 1: The effects of Syzygium cerasoideum extracts on Oral Glucose Tolerance Test. Values expressed as average

Group-I: Control, Group-II: MESC 200mg/kg, Group-III: MESC 400mg/kg, Group-IV: CESC 200mg/kg, Group-V: CESC 400mg/kg, n-06, total duration: 2 hrs

B. Antidiabetic/anti-obesity activity in c57bl/6 j mice

Table 2: The effects of Syzygium cerasoideum extracts on high-fat eating regimen instigated heftiness and diabetes in C57BL/6 J mice

Groups	Dose	Bodyweight
Groups	Dose	Day 0	Day 7	Day 14	Day 21	Day 28
Group-I	control	18.5±0.43	19.5±0.43	20.6±0.33	22.0± 0.45^*	23.1± 0.31^**
Group-II	HFD	18.3±0.33	19.6±0.42	21.8±0.40^*	23.1±0.48^**	26.6±0.67^**
Group-III	MESC 200mg/kg	18.1±0.31	18.6±0.49	19.6±0.67	20.3±0.61	21.5±0.43^*
Group-IV	MESC 400mg/kg	18.5±0.43	19.5±0.56	19.3±0.61	19.0±0.58	19.5±0.43
Group-V	CESC 200mg/kg	18.0±0.37	19.0±0.52	19.3±0.56	20.3±0.49	20.5±0.34
Group-VI	CESC 400mg/kg	18.3±0.42	19.5±0.50	19.1±0.40	20.0±0.73	20.1±0.54

^*p˂0.05 when compared with the control, ^*p˂0.05 when compared with the HFD

Values are expressed as Mean± SEM. All the data were statistically treated with Dunnet Multiple comparison test post one way ANOVA using graph pad version 5.0

Figure 2: The effects of Syzygium cerasoideum extracts on high-fat eating regimen instigated heftiness and diabetes in C57BL/6 J mice Values expressed as average

Table 3: The effects of Syzygium cerasoideum extracts on Glucose and Triglycerides levels high-fat eating regimen instigated heftiness and diabetes in C57BL/6 J mice

Groups	Dose	Glucose in mg/dl	Triglyceride mg/dl
Group-I	control	98.9±1.72⁺⁺	77.3±1.20⁺⁺
Group-II	HFD	158±2.92^**	102±2.66^**
Group-III	MESC 200mg/kg	119±1.99^**++	97.2±1.66^**
Group-IV	MESC 400mg/kg	107±2.52⁺⁺	94.2±2.09^**
Group-V	CESC 200mg/kg	110±3.14^*++	95.4±2.89^**
Group-VI	CESC 400mg/kg	98.5±1.98⁺⁺	89.8±1.87^**++

+P˂0.05 is considered to be significant when compared to HFD

Values are expressed as Mean± SEM. All the data were statistically treated with Dunnet Multiple comparison test post one way ANOVA using graph pad version 5.0

Figure 3: The effects of Syzygium cerasoideum extracts on Glucose and Triglycerides levels in high-fat eating regimen instigated heftiness and diabetes in C57BL/6 J mice Values expressed as average

DISCUSSION:

Consequences of the impact of reviewed portions of Syzygium cerasoideum extricates on typical sound rodents and when managed 60min preceding the glucose stacking created critical decrease ( p˂0.05) with MESC at the portion of 400mg/kg delivered a decrease in blood glucose levels at 30min individually, when contrasted with vehicle-treated gathering at 30min.other gatherings were measurably not huge. The high-fat eating routine encouraged C57BL/6 mice demonstrated a noteworthy decrease of body loads on day 21 and 28, glucose and triglyceride levels toward the part of the bargain were measurably critical, *p˂0.05 when contrasted and the control, *p˂0.05 when contrasted and the HFD, shows the antidiabetic and antiobesity movement of the two concentrates. The results have exhibited that MESC 200 and 400mg/kg have Antidiabetic property.,

The C57BL/6 mouse model utilized in the present investigation is considered to firmly impersonate the movement of heftiness and diabetes in human¹⁷. Thusly this model is the most generally utilized and comprehensively acknowledged model, important to abstain from food instigated obesity¹⁹. In writing, it is accounted for that C57BL/6 mice when sustained with HFD not indispensable, create weight, hyperglycemia and hyperinsulinemia alongside hypertension.

The past examination, we found that a few plants could upgrade glucose take-up and furthermore indicated expanded articulation of adiponectin, leptin, PPAR-γ, and GLUT-4gene articulation in vitro^20,21.In writing, the job of these markers in the advancement of diabetes and insulin has been all around recorded. To put it plainly, PPAR-γ is an atomic factor which ties with the PPAR-responsive element(PPRE) and builds the adiponectin advertiser movement in adipocytes²². Adiponectin is the most gigantically communicated adipokine and is notable for its insulin-sharpening activity; through adiponectin receptors (AdipoR1 and AdipoR2) it initiates AMPK, PPAR-α, and other obscure pathways, to upgrade insulin affectability. In this manner, adiponectin has a positive relationship with insulin sensitivity²³. In skeletal muscles, adiponectin builds the statement of CD36 and acyl-coenzyme-An oxidase, and in this manner lessens the free unsaturated fat and triglyceride levels, which thus add to improved insulin flagging transduction²⁴. Further, adiponectin likewise invigorates phosphorylation of acetyl coenzyme-A carboxylase(ACC) and actives AMPK, which results in upgrade glucose take-up, expanded unsaturated fat burning, diminished gluconeogenesis, and every one of these progressions together adds to decreased blood glucose level²⁵. In addition, leptin is one more boss adipocytokine known to assume an essential job in keeping up the body weight, digestion and conceptive functions²⁶, leptin to a great extent exists in the focal sensory system and is known for its prevalent job in directing nourishment utilization, explicitly lesser degrees of leptin increment sustenance utilization; it is all around showed in leptin knockout animals²⁷, and furthermore it is notable that large people have exceptionally low leptin levels when contrasted with solid volunteers²⁸.

In concurrence with the writing reports, in the present investigation, the HFD control animals demonstrated a critical increment in body weight, blood glucose and insulin levels contrasted with control. MESC (200 and 400mg/kg) treatment lightened all these metabolic changes like body weight, blood glucose and hyperlipidemia that happened due to HFD.

CONCLUSION:

With the outcomes of the present study, we can conclude that MESC possesses antidiabetic by ameliorating the HFD induced body weight gain, hyperglycemia.

ACKNOWLEDGEMENT:

We are thankful to Dayananda Sagar University and Oxbridge College of Pharmacy, Bengaluru for their support.

CONFLICTS OF INTEREST:

The authors declare no potential conflicts of interest concerning the research, authorship, and/or publication of this paper.

REFERENCES:

1. Barness L.A., Opitz J.M., Gilbert-Barness E. Obesity: Genetic, molecular, and environmental aspects. Am. J. Med. Genet. Part A. 2007; 143: 3016–3034.

2. Friedman, J.M. Obesity in the new millennium. Nature 2000; 404: 632–634.

3. Hotamisligil, G.S. The role of TNFα and TNF receptors in obesity and insulin resistance. J. Intern. Med. 1999; 245: 621–625.

4. Shimomura, I. et al. Enhanced expression of PAI-1 in visceral fat: a possible contributor to vascular disease in obesity. Nature Med. 1996; 2: 800–803.

5. White, R.T. et al. Human adipsin is identical to complement factor D and is expressed at high levels in adipose tissue. J. Biol. Chem. 1992; 267: 9210–9213.

6. Steppan, C.M. et al. The hormone resistin links obesity to diabetes. Nature 2012; 409:307–312.

7. Powell A.G., Apovian C.M., Aronne L.J. New drug targets for the treatment of obesity. Clin. Pharmacol. Ther. 2011; 90: 40–51.

8. Lim D.W., Kim Y.T., Jang Y.J., Kim Y.O., Han D.S. Anti-obesity effect of Artemisia capillaris extracts in high-fat diet-induced obese rats. Molecules. 2013;18: 9241–9252.

9. Yun J.W. Possible anti-obesity therapeutics from nature—A review. Phytochemistry. 2010; 71:1625–1641.

10. Haron, N.W.; Moore, D.M,;Harbome, J.B distribution and taxonomic significance of flavonoids in the genus Eugenia (myrtaceae) Biochemical systematic and ecology Vol.20, 226-268 (1992).

11. Mehta RM Pharmaceutics-1.1^st edn, Vallabha Prakashan, Delhi. (1994) 138-140.

12. Sadik S, Geetha KM and Vasia: In-vivo antioxidant and antihyperlipidemic activity of Syzygium cerasoideum extracts in rats. Int J Pharm Sci & Res 2019; 10(6): 1000-1009.

13. Khandelwal KR. Practical pharmacognosy techniques and experiments. 14^th edn, Nirali Prakashan, Pune. (2005) 149-156.

14. Bartoli, G.P. Fra, G.P. Carnevale Schianca. The oral glucose tolerance test. European journal of internal medicine 2011; 22: 8-12.

15. Prakasam, A., Sethupathy, S., Pugalendi, K. Effect of Casearia esculent root extract on blood glucose and plasma antioxidant status in streptozotocin-diabetic rats. Polish Journal of Pharmacology .2003; 55: 43–49.

16. Surwit RS, Kuhn CM, Cochrane C, Mc Cubbin JA, Feinglos MN. Diet-induced type II diabetes in C57BL/6J mice. Diabetes 1988; 37 : 1163-1167.

17. Speakman J, Hambly C, Mitchell S, Krol E. Animal models of obesity. Obes Rev 2007; 8: 55-61.

18. Winzell MS, Ahren B. The high-fat diet-fed mouse: a model for studying mechanisms and treatment of impaired glucose tolerance and type 2 diabetes. Diabetes 2004; 53 : 215-219.

19. Johnson PR, Greenwood MR, Horwitz BA, Stern JS. Animal models of obesity genetic aspects. Annu Rev Nutr 1991; 11: 325-353.

20. Mahesh Kumar P, Venkataranganna MV, Manjunath K, Viswanatha GL, Ashok G. Methanolic extract of Momordica cymbalaria enhances glucose uptake in L6 myotubes in vitro by up-regulating PPAR-γ and GLUT-4. Chin J Nat Med 2014; 12: 895-900.

21. Mahesh Kumar P, Venkataranganna MV, Manjunath K, Viswanatha GL, Ashok G. Methanolic fruit extract of Momordica cymbalaria alleviates insulin resistance by enhancing expression of adiponectin and leptin in vitro. World J Pharm Pharm Sci 2016; 5: 1925-1935.

22. Iwaki M, Matsuda M, Maeda N, Funahashi T, Matsuzawa Y, Makishima M, Shimomura I. Induction of adiponectin, a fat-derived antidiabetic and antiatherogenic factor, by nuclear receptors. Diabetes2003; 52: 1655-1663.

23. Bouskila M, Panjvani UB, Scherer PE. Adiponectin: a relevant player in PPAR-γ agonist-mediated improvements in hepatic insulin sensitivity. Int J Obes 2005; 1: S17-S23.

24. Ronti T, Lupatteli G, Mannarino E. The endocrine function of adipose tissue: an update. Clin. Endocrinol 2006; 64: 355-365.

25. Zheng F, Zhang S, Lu W, Wu F, Yin X, Yu D, Pan Q, Li H. Regulation of insulin resistance and adiponectin signalling in adipose tissue by liver X receptor activation highlights a cross-talk with PPAR-γ. PLoS One 2014; 9: e101269.

26. Flier JS. Obesity wars: molecular progress confronts an expanding epidemic. Cell 2004; 23: 337-350.

27. Zhang Y, Proenca R, Maffei M, Barone M, Leopold L, Friedman JM. Positional cloning of the mouse obese gene and its human homologue. Nature 1994; 372: 425-432.

28. Farooqi IS, Matarese G, Lord GM. Beneficial effects of leptin on obesity, T cell hyporesponsiveness, and neuroendocrine/metabolic dysfunction of human congenital leptin deficiency. J Clin Invest 2002; 110:1093-1103.

Received on 29.07.2019 Modified on 20.08.2019

Research J. Pharm. and Tech 2020; 13(3): 1297-1302.

DOI: 10.5958/0974-360X.2020.00239.5