## 4 Discussion  
A major issue considered in the present study was whether the aqueous extract of ripe _Carissa carandas_ Linn. (CCA) could exert an antidiabetic effect or not. The outcomes of this study provide evidence to suggest that the antidiabetic effect of CCA is possibly mediated via reducing inflammation and insulin resistance using the in vitro model and in vivo model of T2 DM rats. The percentage yield of _C. carandas_ dried fruits extracted in an aqueous solution (CCA) was 38.8%, which is comparable to a previous study by [PERSON] et al. with a yield of 32.0% by using aqueous extraction [20]. The amount of total phenolic and flavonoid contents in the CCA extract (18.23 \(\pm\) 0.4 mg GAE/g extract and 12.04 \(\pm\) 2.7 mg RUE/g extract, respectively) were higher than the aqueous extract of _C. carandas_ fruit reported by [PERSON] et al. (5.3 \(\pm\) 3.8 mg GAE/g extract and 2.7 \(\pm\) 1.5 mg CE/g extract, respectively) [20]. The ripe fruit of _C. carandas_ is a rich source of the anthocyanin pigment. The major anthocyanin observed in this study was cyanidin-3-O-galactoside. This result confirms a previous report by [PERSON] et al., who found cyanidin-3-O-galactoside  
Figure 8: Photomicrographs of pancreatic tissues: normal control group (**A**,**G**); normal rats treated with 400 mg/kg BW of CCA (**B**,**H**); diabetic control group (**C**,**I**); 200 mg/kg BW of CCA (**D**,**J**); 400 mg/kg BW of CCA (**E**,**K**) and metforminin (**F**,**L**) given to diabetic rats. The black-boxed areas in A-F are magnified in G–L AC: acinar cells; IL: islets of Langerhans cells. Cytoplasmic vacuolar degeneration (red arrow); Pyknotic nuclei (yellow arrow head); hematoxylin and eosin; (**A–F**) (40\(\times\)), (**G–L**) (100\(\times\)). to be the major anthocyanin compound in the ripe _C. carandas_ fruit [10]. This finding was in contrast to the earlier reports. Previously, [PERSON] et al. reported that cyanidin-3-O-galactoside is a major pigment found in the ripe _C. carandas_ fruit [11]. Moreover, the ripe _C. carandas_ fruit has been reported by [PERSON] et al. to have three anthocyanin pigments. The major anthocyanin of the _C. carandas_ ripe fruit was cyanidin-3-O-glucoside, followed by peonidin-3-O-glucose and malvidin-3-O-glucose [12]. In addition, the _C. carandas_ ripe fruit was previously reported to contain eight anthocyanins, which are cyanidin-3-O-glucose, cyanidin-3-O-galactoside, delphinidin-3-O-galactoside, delphinidin-3-O-glucose, delphinidin-3-O-rutinoside, 5-carboxypyanopelargonidin-3-beta-glucopyranoside, delphinidin, and peonidin-3-O-arabinoside, respectively [21]. The observed differences in phytochemical contents in the _C. carandas_ fruit in this study might be due to several factors, including fruit ripeness, geographical locations, climatic conditions, and extraction methods [22; 23; 24; 25]. An imbalance between the production of free radicals and the ability of the antioxidant system in the human body results in oxidative stress. Moreover, studies revealed that oxidative stress upregulates inflammatory mediators and also induces cellular or tissue damage, which leads to the pathogenesis of several diseases, such as insulin resistance, T2 DM, and cardiovascular diseases [26]. The results from this study showed the abilities of the CCA extract to scavenge ABTS free radicals in a dose-dependent manner. Since the higher total antioxidant capacity of the diet has been found to decrease the risk of T2 DM and metabolic diseases, the antioxidant activity of the CCA extract might be involved with its anti-inflammation and anti-insulin resistance [27; 28]. A previous study found more macrophage migration into adipose tissue in obese mice than in lean mice [2]. Macrophages can activate the nitric oxide (NO) and the pro-inflammatory cytokines, including IL-1\(\beta\), IL-6, and TNF-\(\alpha\) production and secretion, resulting in a low-grade chronic inflammation condition in adipose tissue which can promote insulin resistance in adipocytes [3].