COMPARISON

OF

ANTI

INFLAMMATORY

EFFECT

BETWEEN

ANTHOCYANINS EXTRACT OF BLACK RICE (Oryza sativa L) AND PURPLE

SWEET POTATO (Ipomea batatas poiret) IN TYPE 2 DIABETIC RATS

Mislachah Nuril
Faculty of Agricultural Technology, Universitas Gadjah Mada, Indonesia
Mary Astuti**
Faculty of Agricultural Technology, Universitas Gadjah Mada, Indonesia
Sunarti
Faculty of Medicine, Universitas Gadjah Mada, Indonesia

**corresponding author
Phone number : +62 85729707222
Email

: ana_food@yahoo.com; mislachahnuril@gmail.com

Keywords:
anthocyanin extract
black rice
purple sweet potato
hyperglycemia
anti-inflammatory.

ABSTRACT
Diabetes Mellitus is a metabolic disease due to disruption of insulin and more exacerbated by
oxidative stress and inflammation. Animal models induced hyperglycemia with streptozotocin
(STZ) and nicotinamide (NA) can cause inflammation and damage to pancreatic cells,
thereby reducing the amount of insulin secreted. Anthocyanin is a polyphenolic compound
that can act as antioxidants and anti-inflammatory. This research is an experimental research
by testing the effect of anthocyanin extract of black rice as well as the provision of
anthocyanin extract of purple sweet potato in rat of hyperglycemia to decrease proinflammatory cytokines TNF- and IL-6. The results showed that black rice anthocyanin
extract and anthocyanins extract of purple sweet potato in rats for 6 weeks hyperglycemia
give positive effect. It decreased pro-inflammatory cytokines levels until the end of the
intervention.
INTRODUCTION
Diabetes Mellitus (DM) is a metabolic disease, which is characterized by increased blood
glucose levels (hyperglycemia). There are two types of diabetes, namely: is insulindependent diabetes mellitus (type 1 diabetes) and non-insulin-dependent diabetes mellitus
(type 2 diabetes). Type 2 diabetes is associated with insulin resistance and relative insulin
deficiency and characterized by high blood glucose levels.
Insulin resistance is a condition associated with organ failure to respond targets
normally the activity of the hormone insulin. Insulin resistance can be induced due to
inflammatory factors. Cytokines have an important role in the inflammatory process
(Gerszten RE, et al. 1999). TNF- is an inflammatory cytokine that is involved in
metabolic disorders, including insulin resistance (Kern PA, et al. 1995). In addition to TNF, IL-6 is also an inflammatory cytokine that can alter insulin sensitivity by triggering
different insulin signaling pathway (Bastard JP et al, 2000). Insulin resistance can also be

induced by oxidative stress. Oxidative stress can lead to apoptosis of pancreatic cells
resulting in insulin resistance (Evans, 2002; Oprescu, 2007).
Anthocyanins are believed to have very good antioxidant effects, because it can inhibit a
variety of free radicals causing oxidative stress. Anthocyanin antioxidant properties lies in
its ability to donate an electron or hydrogen atom transfer from the hydroxyl group to the
free radicals, so that free radicals are relatively stable and will not attack the other
components in the cell or the cell membrane (Prior, 2003).
The purpose of this study is to examine the effect of anthocyanin extracts of black rice
compared with anthocyanin extract of purple sweet potato to decrease inflammation in
rat models of hyperglycemia.
MATERIAL AND METHOD
Samples

of the study using experimental animals such as

male strain rats

(Rattusnorvegicus) of Sprague-Dawley, 3 months old, weighing between 200-250 grams.

Experimental animals were divided into 4 groups, control (C), diabetes mellitus group
(DM), diabetes mellitus group with anthocyanin extract of black rice (BR), diabetes mellitus
group with anthocyanin extract of purple sweet potato (PSP).
Induction of diabetes mellitus used STZ (streptozotocin) (65 mg/kg) and NA (nicotinamide)
(230 mg/kg body weight) intraperitoneally (IP). Anthocyanin extract was given for 6 weeks
with different doses, at week 1 to 3, anthocyanin given was 13.5 mg/kg/day and then at
week 4 to 6, it was given additional doses greater than 5 times greater ie 67.5 mg/kg/day.
Blood glucose calculation was performed using GOD-PAP enzymatic method, proinflammatory cytokines TNF- and IL-6 using ELISA Kit.
RESULTS
Blood Glucose Levels
Testing of blood glucose levels is an important analysis of the conditions of hyperglycemia.

Changes in blood glucose in each group during the maintenance period can be seen in Table
1.
(Table 1)
After induction using STZ and NA, partly rat pancreatic beta cells would be damaged and
was unable to produce insulin. This resulted in increased blood glucose levels of rat week 0.
Treated group of rat which received the extract of black rice anthocyanin or purple sweet
potato showed that blood glucose levels decreased until the end of the intervention. This
showed the positive effect of anthocyanin extract to decrease blood glucose. Guo et al.
(2007) reported a decline in insulin resistance (HOMA-IR impairment amounted to 27.67%)
due to the provision of anthocyanins from black rice and black soybeans that blood
glucose also decreased. Takikawa et al., (2010) also reported the repair mechanisms of
hyperglycemia conditions as indicated by a decrease in blood glucose levels and improved
insulin sensitivity after administration of bilberry extract in diabetic rats for 4 weeks.
Increased doses of 67.5 mg of anthocyanin extract did not have a significant influence
with the administration of a 13.5 mg dose of anthocyanin extract. According to Hermawati
(2013), administration of purple sweet potato anthocyanin extract at a dose of 50 mg/kg
and 100 mg/kg for 35 days was capable of lowering blood glucose hyperglycemic rats
respectively 33.90% and 38.83%.
Pro-inflammatory cytokines TNF- and IL-6
(Table 2)
Levels of TNF- and IL-6 on diabetic control rat showed a high value and continue
to increase until the end of the week intervention. While on treatment of anthocyanin
extracts of black rice and purple sweet potato, inflammation showed a significant
improvement status by decreasing the amount of secreted cytokines. TNF- and IL-6 has a
close relationship with the occurrence of insulin resistance and high levels of glucose.

DISCUSSION
High glucose levels occur due to insulin resistance which can be induced due to
inflammatory factors. Inflammation was defined as a phenomenon that occurs due to
pathological stimuli and wounding network (Cone, JB. 2001). The inflammatory response is
simply divided into two phases: acute phase usually followed by a chronic phase. The
purpose of inflammation is to destroy and eliminate the agent destroyer. Cytokines have an
important role in the inflammatory process (Gerszten RE et al., 1999). Increased levels of
cytokines in inflammatory markers were also involved in metabolic disorders (Fasshauer
and Paschke R, 2003).
TNF- and IL-6 could alter insulin sensitivity by triggering different insulin signaling
pathway (Bastard JP, et al., 2000). These cytokines can stimulate phosphorylation of serine
residues in the insulin receptor substrate-1 (IRS-1), so it prevented activation of insulin
signaling and resulted in insulin resistance (Fasshauer and Paschke R, 2003). This was in line
with the results of the correlation calculations, which showed that blood glucose levels
positively correlated with the levels of TNF- and IL-6. Ford ES (2003) and Yudkin et al.,
(1999), also found IL-6 and TNF- were positively correlated with the development of
insulin resistance.
TNF- could alter insulin sensitivity in several different ways, such as by attenuation of
insulin receptor signaling pathway (Hotamisligil, 2006), decreases GLUT-4 in adipocytes
(Stephens JM, et al., 1997), and suppresses adiponectin (Wang et al., 2006). High Free Fatty
Acids that cause oxidative stress also contributed to insulin resistance by activating Toll-Like
Receptor (TLR) and resulted in cytokine response or by directly activating June Nterminal Kinase (JNK) which could stimulate phosphorylation of serine residue found on
the IRS -1 (Hotamisligil, 2008).

CONCLUSION
Based on the results that have been described, the conclusions that can be drawn include: (1)
black rice anthocyanin extract and purple sweet potato has antiinflamatory effect of being
able to lower blood glucose levels, and reduce inflammatory markers TNF- and IL-6, (2)
using different dose of anthocyanin extract without washingout has been able to provide
decrease diabetic and inflammatory conditions on blood glucose levels, inflammatory
markers TNF- and IL-6.
ACKNOWLEDGMENTS
The author would like to thank the directorate of research and community service, the
ministry of research, technology and higher education, which has funded this research.
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