Antidiabetic, antidyslipidemia, and renoprotector potency of butterfly pea flower extract (Clitorea ternatea L.) in diabetes mellitus and dyslipidemia rats model

Abstract

Background: Diabetes mellitus (DM) is a long-term condition marked by high blood glucose levels caused by insulin resistance which will lead to complications of other diseases such as dyslipidemia, which also affects the health of the liver and kidneys. Butterfly pea flower (Clitorea ternatea L.) has phenolic and flavonoid compounds which have the potential as herbal medicines for antidiabetics.

Aim: The purpose of this study is to examine the potential of butterfly pea flower extract (BPE) as an antidiabetic, anti-dyslipidemia, and renoprotection.

Methods: In vivo test was performed on Sprague Dawley rats (Rattus norvegicus L.) induced by Streptozotocin-Nicotinamide and High Fat Diet-Propylthiouracil as models of DM and dyslipidemia, and BPE was administered orally (200, 400, and 800 mg/kg BW) for 28 days. glutathione peroxidase (GSH-Px), glutathione S-transferase (GST), tumor necrosis factor-α (TNF-α), nuclear factor-kappa beta (NF-kB), alkaline phosphatase (ALP), liver albumin levels, serum blood urea nitrogen (BUN), serum creatinine, and serum uric acid (UA), were measured by ELISA and colorimetry methods.

Results: Treatment of BPE 800 mg/kg BW increased levels of GSH-Px, GST, albumin, and serum protein. BPE decreased TNF-α, NF-kB, and ALP. BPE also decreased BUN, serum CR, and serum UA.

Conclusion: BPE has the potential to be used as a drug alternative for the treatment of DM and dyslipidemia as well as a hepatoprotective and renoprotective agent.

Keywords: Antidiabetic; Clitorea ternatea L.; Diabetes mellitus; Dyslipidemia; Renoprotective.

Fig. 1.. Effects of various treatments on GSH-Px activity and GST activity in DM and dyslipidemia rat models; (A) GSHPx activity, (B) GST activity. *Data are presented as means ± SD of four repetitions. The different superscript marks a, ab, bc, cd on GSH-Px activity (U) and a, ab, bc, c on GST activity (U/ml) showed a significant difference (p < 0.05). Group I: negative control (aquadest), Group II: positive control (HFD, PTU, STZ 60 mg/kg BW, NA 120 mg/kg BW), Group III: positive control + BPE 200 mg/kg BW/day, Group IV: positive control + BPE 400 mg/kg BW/day, Group V: positive control + BPE 800 mg/kg BW/day, Group VI: positive control + Simvastatin 0.9 mg/kg BW, Group VII: positive control + Glibenclamide 0.45 mg/kg BW, Group VIII: positive control + Glibenclamide 0.45 mg/kg BW + Simvastatin 0.9 mg/kg BW.

Fig. 2.. Effects of various treatments on TNF-α levels in DM and dyslipidemia rat models. *Data are presented as means ± SD of four repetitions. The different superscript marks a, b, c, d, de, e on TNF-α level (pg/ml, pg/mg prot) showed a significant difference among treatments (p < 0.05). Group I: negative control (aquadest), Group II: positive control (HFD, PTU, STZ 60 mg/kg BW, NA 120 mg/kg BW), Group III: positive control + BPE 200 mg/kg BW/day, Group IV: positive control + BPE 400 mg/kg BW/day, Group V: positive control + BPE 800 mg/kg BW/day, Group VI: positive control + Simvastatin 0.9 mg/kg BW, Group VII: positive control + Glibenclamide 0.45 mg/kg BW, Group VIII: positive control + Glibenclamide 0.45 mg/kg BW + Simvastatin 0.9 mg/kg BW.

Fig. 3.. Effects of various treatments on NF-kB levels in DM and dyslipidemia rats model. *Data are presented as means ± SD of four repetitions. The different superscript marks a, ab, bc, c on NF-kB levels (pg/ml) and a, ab, c, d on NF-kB levels (pg/mg prot) showed a significant difference among treatment (p < 0.05). Group I: negative control (aquadest), Group II: positive control (HFD, PTU, STZ 60 mg/kg BW, NA 120 mg/kg BW), Group III: positive control + BPE 200 mg/kg BW/day, Group IV: positive control + BPE 400 mg/kg BW/day, Group V: positive control + BPE 800 mg/kg BW/day, Group VI: positive control + Simvastatin 0.9 mg/kg BW, Group VII: positive control + Glibenclamide 0.45 mg/kg BW, Group VIII: positive control + Glibenclamide 0.45 mg/kg BW + Simvastatin 0.9 mg/kg BW.

Fig. 4.. Effects of various treatments on ALP activity in DM and dyslipidemia rats model. *Data are presented as means ± SD of four repetitions. Different superscript marks a, ab, c, cd, d, de, e on ALP activity (King Unit/100 ml) and a, ab, bc, c, d, e, f on ALP activity (King Unit/mg protein) showed a significant difference among treatments (p < 0.05). Group I: negative control (aquadest), Group II: positive control (HFD, PTU, STZ 60 mg/kg BW, NA 120 mg/kg BW), Group III: positive control + BPE 200 mg/kg BW/day, Group IV: positive control + BPE 400 mg/kg BW/day, Group V: positive control + BPE 800 mg/kg BW/day, Group VI: positive control + Simvastatin 0.9 mg/kg BW, Group VII: positive control + Glibenclamide 0.45 mg/kg BW, Group VIII: positive control + Glibenclamide 0.45 mg/kg BW + Simvastatin 0.9 mg/kg BW.

Fig. 5.. Effects of various treatments on albumin levels in DM and dyslipidemia rat models. *Data are presented as means ± SD of four repetitions. The different superscript marks a, b, c, d for albumin content (g/l) and a, b, bc, d for albumin content (g/mg prot) showed significant differences among treatments (p < 0.05). Group I: negative control (aquadest), Group II: positive control (HFD, PTU, STZ 60 mg/kg BW, NA 120 mg/kg BW), Group III: positive control + BPE 200 mg/kg BW/day, Group IV: positive control + BPE 400 mg/kg BW/day, Group V: positive control + BPE 800 mg/kg BW/day, Group VI: positive control + Simvastatin 0.9 mg/kg BW, Group VII: positive control + Glibenclamide 0.45 mg/kg BW, Group VIII: positive control + Glibenclamide 0.45 mg/kg BW + Simvastatin 0.9 mg/kg BW.

Fig. 6.. Effects of BPE on protein content in DM and Dyslipidemia Rats. * The data are shown as mean ± standard deviation from 4 repetitions. Different superscript marks A, B, BC, and C on protein content day 14 (μmol/l) and a, b, bc, and c on protein content day 28 (mmol/l) showed significant differences (p < 0.05). Group I: negative control (aquadest), Group II: positive control (HFD, PTU, STZ 60 mg/kg BW, NA 120 mg/kg BW), Group III: Positive control + BPE 200 mg/kg BW/day, Group IV: Positive control + BPE 400 mg/kg BW/day, Group V: Positive control + BPE 800 mg/kg BW/day, Group VI: Positive control + Simvastatin 0.9 mg/kg BW, Group VII: Positive control + Glibenclamide 0.45 mg/kg BW, Group VIII: positive control + Glibenclamide 0.45 mg/kg BW + Simvastatin 0.9 mg/kg BW.

Fig. 7.. Effects of BPE on BUN, Creatinin, and UA content in DM and Dyslipidemia rats; (A) BUN, (B) Creatinin, (C) UA. * The data are shown as mean ± standard deviation from 4 repetitions. The different superscript marks A, AB, and B on BUN Content day 14 (μmol/l) and a, ab, and b on BUN content day 28 (mmol/l) showed significant differences (p < 0.05). Group I: negative control (aquadest), Group II: positive control (HFD, PTU, STZ 60 mg/kg BW, NA 120 mg/kg BW), Group III: Positive control + BPE 200 mg/kg BW/day, Group IV: Positive control + BPE 400 mg/kg BW/day, Group V: Positive control + BPE 800 mg/kg BW/day, Group VI: Positive control + Simvastatin 0.9 mg/kg BW, Group VII: Positive control + Glibenclamide 0.45 mg/kg BW, Group VIII: positive control + Glibenclamide 0.45 mg/kg BW + Simvastatin 0.9 mg/kg BW.

Fig. 8.. Proposed mechanism of BPE as hepatoprotector and renoprotector in DM and Dyslipidemia rat model. In diabetic conditions, ROS in the body such as H₂O₂ are produced in excess. Normally this H₂O₂ can be overcome by the activity of the GPX enzyme to become H₂O through the oxidation of GSH to (Glutathione Disulfide) GSSG. However, the large number of H₂O₂ molecules disrupts the activity of the GSH-Px enzyme. On the other hand, other antioxidant enzymes, namely GST, can also convert GSH and xenobiotics, namely foreign substances that are not beneficial to the body, into safer Glutathione S-Conjugates. The presence of free radicals can also activate NFkB which causes the production of TNF-α which has an impact on inflammation. The presence of oxidative stress and inflammation in cells over time will develop into damage to organs such as the liver and kidneys. Liver damage is characterized by increased ALP enzymes, as well as decreased protein and albumin. In the kidney, inflammation and oxidative stress have an impact on decreasing the GFR which causes BUN, SCr, and UA in the blood cannot be excreted. Treatment with BPE can neutralize H₂O₂ in cells by increasing GSH-Px and GST enzyme activity, also downregulating NF-kB and TNF-a, as well as reducing the risk of liver damage by reducing ALP enzyme expression and increasing total protein and albumin. BPE also increases GFR so that BUN, SCr, and UA can be excreted properly.