Mechanism of Action of Natural Dipeptidyl Peptidase-IV Inhibitors (Berberine and Mangiferin) in Experimentally Induced Diabetes with Metabolic Syndrome.

Background: Berberine (BER) and mangiferin are known natural dipeptidyl peptidase (DPP-IV) inhibitors. Hence, the study was designed to elucidate the mechanism of action of natural DPP-IV inhibitors (BER and MNG) in experimentally induced diabetes with metabolic syndrome. Aim: The aim of this study was to observe mechanism through which natural DPP-IV inhibitor works in diabetes with metabolic syndrome rat model. Materials and Methods: Wistar rats were fed high-fat diet for 10 weeks and challenged with streptozotocin (STZ) (40 mg/kg) at the 3rd week (high-fat diabetic control [HF-DC] group). After the confirmation of metabolic syndrome in the setting of diabetes, monotherapy (metformin [MET], vildagliptin [VIL], BER, and MNG) and combination (MET + VIL, MET + BER, and MET + MNG) therapy was orally fed to these rats from the 4th to 10th weeks. Results: Insulin resistance (IR) was seen in the HF-DC group as indicated by raised homeostasis model assessment of IR (HOMA-IR) in HF-DC group as compared with normal control (NC) groups. The treatment groups reduced IR as shown by a decrease in HOMA-IR as compared with HF-DC group rats. The marked reduction (P < 0.001) of beta-cell function was observed in the HF-DC group as a reduced level of HOMA for beta-cell function (HOMA-ß) was found as compared with the NC group. Increases in HOMA-ß as compared to the HFDC group were observed in the therapy groups. The treatment group significantly reduced cholesterol and atherogenic index. The treatment group showed significant preservation of beta-cell mass as per immunohistochemistry and significant anti-apoptotic activity as per Terminal Deoxyribonucleotidyl Transferase-Mediated dUTP Nick End Labeling assay report. The treated rats significantly (P < 0.05) reduced high-sensitivity C-reactive protein. Lipid peroxidation (thiobarbituric acid reactive substances) marker (P < 0.001) was significantly reduced in the treatment group. Conclusion: The natural DPP-IV inhibitors BER and MNG treatment showed beneficial effects on various components of metabolic syndrome.

Antidiabetic activity of Commiphora mukul and Phyllanthus emblica and Computational analysis for the identification of active principles with dipeptidyl peptidase IV inhibitory activity.

The medicinal plants may serve as natural alternatives to synthetic antidiabetic medications such as dipeptidyl peptidase-IV (DPP-IV) inhibitors, which are commonly prescribed in clinical practise. The medicinal plants: Commiphora mukul and Phyllanthus emblica have considerable DPP-IV inhibitory efficacy, according to our findings. The present study is an extension of the previous study conducted in our laboratory and was designed to confirm the antidiabetic effects of C. mukul and P. emblica in the streptozotocin diabetes model and elucidate the active principles responsible for DPP-IV inhibition. C. mukul (Guggul) and P. emblica (Amla) have the ability to inhibit DPP-IV and have anti-diabetic properties in a Type 2 diabetes mellitus experimental model. The binding sites and affinity of the active principles of C. mukul (Gluggusterone E, Gluggusterone Z) and P. emblica (Pzrogallol, beta-glucogallin, and gallic acid) responsible for DPP-IV enzyme inhibition were identified using in silico studies and compared to Vildagliptin, a synthetic DPP-IV inhibitor. The Vildagliptin and therapy groups had significantly lower glycated hemoglobin and DPP-IV levels. The anti-diabetic effect of C. mukul and P. emblica is due to their DPP-IV inhibitory action. The DPP-IV inhibitory action of Gluggusterone E, Gluggusterone Z, and beta-Glucogallin was found to be superior to Vildagliptin in docking tests.

Dipeptidyl peptidase IV Inhibitory activity of Terminalia arjuna attributes to its cardioprotective effects in experimental diabetes: In silico, in vitro and in vivo analyses.

BACKGROUND: The marketed synthetic (Dipeptidyl peptidase-IV) DPP-IV Inhibitors are expensive antidiabetic drugs and have been reported to cause unacceptable adverse effects such as pancreatitis, angioedema, thyroid and pancreatic cancers. In this scenario research to develop novel DPP-IV Inhibitors from alternative sources is the need of the hour. HYPOTHESIS/PURPOSE: Terminalia arjuna, a medicinal herb with antidiabetic and cardioprotective activities may represent a natural DPP-IV Inhibitor, the DPP-IV Inhibitory activity of which may translate into demonstrable therapeutic benefits in setting of diabetes with cardiovascular co-morbidities. STUDY DESIGN: The study type used for the present study was an experimental (In vitro, In vivo and In silico) design. METHOD: The DPP-IV Inhibitory, antidiabetic and cardioprotective effects of Terminalia arjuna was evaluated in the experimental model of myocardial infarction co-existing with diabetes. To determine the active principle of Terminalia arjuna responsible for DPP-IV Inhibitory activity, the crystal structure of DPP-IV was considered as receptor which was docked against Arjunetin, Arjungenin, Arjunic acid, Arjunone, Ellagic acid, Gallic acid, Sitagliptin and Vildagliptin. The binding sites as well as affinity of various active ingredients of Terminalia arjuna for DPP- IV enzyme was elucidated using in silico studies and compared to Vildagliptin. RESULTS: Terminalia arjuna demonstrated significant DPP-IV Inhibitory, antidiabetic (significant reduction in HbA1C) and cardioprotective effects (restoration of myocardial CPK-MB) in the experimental model of myocardial infarction co-existing with diabetes. The cardioprotective efficacy correlated to its DPP-IV Inhibitory activity. The active ingredients of Terminalia arjuna (Arjunetin, Arjungenin, Arjunic Acid Arjunone, Ellagic acid and Gallic acid) demonstrated significant inhibition of DPP-IV enzyme. Arjunic acid and Arjunone prefers the active site pocket of DPP-IV enzyme. Compounds like Arjunetin and Vildagliptin prefers to bind near the interface region of the DPP-IV as their biological active forms are homodimer. Sitagliptin binds near the α/ß hydrolase domain. CONCLUSION: The DPP-IV Inhibitory activity of Terminalia arjuna was found to be comparable to Vildagliptin. The DPP-IV Inhibitory activity translated into significant cardioprotective effects in the setting of diabetes. The active ingredient of Terminalia arjuna; Arjunetin, Arjungenin, Ellagic acid and Arjunic acid showed superior DPP-IV Inhibitory activity as compared to synthetic DPP-IV inhibitors (Sitagliptin and Vildagliptin) based on results of docking studies.

Acceptance of Contraceptive Methods Among Postpartum Women in a Tertiary Care Center.

BACKGROUND: In India, a large proportion of women with an unmet need for contraception are within their first year after childbirth. Therefore, concentrating efforts to educate postpartum women on the importance of attending contraceptive clinics could have a proportionally bigger impact on increasing postpartum contraception usage. METHODS: Hundred and seventy-eight (178) women were followed up to determine the proportion of postpartum women who attended the family planning clinics for contraceptive counseling. The reason for non-attendance, choice and effectiveness of contraceptive method selected was determined. RESULTS: Out of 178 postpartum women who were followed up, only 12 (6.8 %) attended the contraceptive clinic. IUD, POPs and Inj-DMPA are the preferred contraceptive methods selected by postpartum women. Hundred percent of the postpartum women who attended contraceptive selected a contraceptive method as compared to only 44 % of the postpartum women who did not attend a contraceptive clinic. Only 29.2 % of these postpartum women selected highly effective contraceptive methods as compared to 83.3 % by the postpartum women who attended family planning clinics. The common reasons cited for not attending contraceptive clinic was found to be time constraint (43.9 %) followed by 'stay far away' (39 %), followed by 'already have information' (9.7 %). CONCLUSIONS: Family planning service that is scheduled to be delivered at the 6-week postpartum is rarely attended. The common reason cited by postpartum women for poor attendance in these family planning clinics was time constraint.

Impact of Structured Counseling on Choice of Contraceptive Method Among Postpartum Women.

BACKGROUND OF THE STUDY: In India, although a number of contraceptive choices are available, the usage of contraceptive methods among postpartum women is rather low. The current study intended to determine the impact of "structured contraception counseling" on women's decision making on selection of contraceptive methods and the reasons behind the selection of a contraceptive method. METHODS: One-hundred-and-seventeen postpartum women in the age group of 18-35 years, requesting contraception, were enrolled in the study. "Structured contraception counseling" was provided using a standardized protocol with balanced and comprehensive education material on the available hormonal and nonhormonal contraceptive methods. Questionnaires with information on the women's pre- and post-counseling contraceptive choice, her perceptions, and the reasons behind her postcounseling decision were filled by the participating women. RESULTS: Maximum women enrolled for the study were in the age group of 21-25 years. In pre-counseling, 36 % postpartum women selected a contraceptive method, 23.1 % a nonhormonal method, and 12.8 % a hormonal method. After "structured contraception counseling," 92.25 % of women chose a contraceptive method. There were significant differences between the women's choices of contraceptive methods in the pre- and post-counseling sessions, respectively [progesterone-only pills (POP): 5.1 vs. 38.46 %, (p < 0.001); injectable-depot medroxy progesterone acetate (DMPA): 2.56 vs. 21.356 %, (p < 0.01); and intra uterine device (IUD): 10.28 vs. 23.92 %, (p < 0.001). 38.46 % chose a POP, 21.36 % injectable-DMPA, and 23.9 % the IUD]. CONCLUSION: "Structured contraception counseling" using standardized protocol resulted in significant improvements in the selection of contraceptive methods by postpartum women.

Natural dipeptidyl peptidase-IV inhibitor mangiferin mitigates diabetes- and metabolic syndrome-induced changes in experimental rats.

BACKGROUND: Mangiferin (MNG) is known to possess antidiabetic and antioxidant activity. However, there is no experimental evidence presently available in the literature with regard to its ameliorating effects on diabetes mellitus coexisting with metabolic syndrome. OBJECTIVE: The present study was designed to evaluate the protective effects of MNG on various components of metabolic syndrome with diabetes as an essential component. MATERIAL AND METHODS: Adult Wistar rats were fed high-fat diets for 10 weeks and challenged with streptozotocin (40 mg/kg) at week three (high-fat diabetic control group). After the confirmation of metabolic syndrome in the setting of diabetes, MNG 40 mg/kg was orally fed to these rats from the fourth to tenth week. RESULTS: The treatment with MNG showed beneficial effects on various components of metabolic syndrome, such as reduced dyslipidemia (decreased triglyceride, total cholesterol, low-density lipoprotein cholesterol, and increased high-density lipoprotein cholesterol) and diabetes mellitus (reduced blood glucose and glycosylated hemoglobin). In addition, an increase in serum insulin, C-peptide, and homeostasis model assessment-ß and a reduction in homeostasis model assessment of insulin resistance-IR were observed in MNG-treated group compared with high-fat diabetic control group. MNG was also found to be cardioprotective (reduction in creatine phosphokinase-MB levels, atherogenic index, high-sensitivity C-reactive protein). Reduction in serum dipeptidyl peptidase-IV levels in the MNG-treated group correlated with improvement in insulin resistance and enhanced ß-cell function. CONCLUSION: The present study has demonstrated antidiabetic, hypolipidemic, and cardioprotective effects of MNG in the setting of diabetes with metabolic syndrome. Thus, MNG has the potential to be developed as a natural alternative to synthetic dipeptidyl peptidase-IV inhibitors beneficial in this comorbid condition.

Cardioprotective effects of rutin via alteration in TNF-α, CRP, and BNP levels coupled with antioxidant effect in STZ-induced diabetic rats.

Diabetic cardiomyopathy (DCM) is a dreadful complication of diabetes responsible for 80 % mortality in diabetic patients, but unfortunately its pharmacotherapy is still incomplete. Rutin is a naturally occurring flavonoid having a long history of use in nutritional supplements for its action against oxidative stress, inflammation, and hyperglycemia, the key players involved in the progression of DCM, but remains unexplored for its role in DCM. This study was conducted to address this lacuna. It was performed in 4-week-old Streptozotocin-induced (45 mg/kg) diabetic rats for a period of 24 weeks to mimic the cardiotoxic effect of chronic hyperglycemia in diabetic patient's heart and to investigate the effect of rutin (50 mg/kg/day) in ameliorating these effects. Heart of the diabetic rats showed altered ECG parameters, reduced total antioxidant capacity, increased inflammatory assault, and degenerative changes. Interestingly, rutin treatment significantly ameliorated these changes with decrease in blood glucose level (p > 0.001), % HbA1c (p > 0.001) and reduced expression of TNF-α (p < 0.001), CRP (p < 0.001), and BNP (p < 0.01) compared to diabetic control rats. In addition, rutin provided significant protection against diabetes associated oxidative stress (p < 0.05), prevented degenerative changes in heart, and improved ECG parameters compared to diabetic control rats. The heart-to-body weight ratio was significantly reduced in rutin treatment group compared to diabetic control rats (p < 0.001). In conclusion, this study implicates that oxidative stress and inflammation are the central players involved in the progression of DCM and rutin ameliorates DCM through its antioxidant and anti-inflammatory actions on heart.

Myocardial Salvaging Effects of Berberine in Experimental Diabetes Co-Existing with Myocardial Infarction.

INTRODUCTION: Berberine, an isoquinoline alkaloid isolated from the Berberis aristata, has been shown to display a wide array of pharmacological activities (hypoglycaemic and hypolipidemic). AIM: The present study was designed to investigate whether these pharmacological properties translate into the cardioprotective effects of Berberine in the setting of diabetes mellitus. MATERIALS AND METHODS: Necessary approval from the Institutional Animal Ethics Committee was taken for the study. Experimental diabetes was produced with single dose of Streptozotocin (STZ): 45mg/kg ip and myocardial infarction was induced by administering Isoproterenol (ISP): 85mg/kg, sc to rats on 35(th) & 36(th) day. After the confirmation of diabetes on 7(th) day (>200mg/dl), Berberine (100 mg/kg) was administered orally to experimental rats from day 8 and continued for 30 days thereafter. Various anti-diabetic (Glucose, HbA1c), cardioprotective (CPK-MB), metabolic (lipid profile), safety {liver function (SGPT, kidney function (Creatinine)} and histopathological indices of injury were evaluated in Healthy Control, Diabetic Control and Berberine treated groups. RESULTS: Administration of STZ-ISP resulted in a significant decrease in body weight (p<0.001), diabetic changes (increase in blood glucose, HbA1c), cardiac injury (leakage of myocardial CPK-MB), altered lipid profile, SGPT, creatinine levels (p<0.001) in the diabetic control group rats as compared to healthy control. Berberine treatment demonstrated significant antidiabetic as well as myocardial salvaging effects as indicated by restoration of blood glucose, HbA1c and CPK-MB levels (p<0.001) compared to diabetic control group. In addition, Berberine favourably modulated the lipid parameters (total cholesterol, triglycerides, HDL, LDL). Subsequent to ISP challenge, histopathological assessment of heart, pancreas and biochemical indices of injury confirmed the cardioprotective effects of Berberine in setting of diabetes. In addition, Berberine was found to be safe to the liver and kidney. CONCLUSION: Berberine treatment produced myocardial salvaging effects in the setting of diabetes challenged with ISP induced myocardial necrosis. Cardioprotection may be attributed to anti-diabetic and hypolipidemic activities.

Genistein ameliorates cardiac inflammation and oxidative stress in streptozotocin-induced diabetic cardiomyopathy in rats.

The present study was undertaken to evaluate the protective effects of genistein against cardiac inflammation and oxidative stress in streptozotocin (STZ) (45 mg/kg body weight)-induced diabetic rats. genistein (300 mg/kg/day) was administered orally for 24 weeks to STZ-induced diabetic rats. The effects of genistein on blood glucose, % glycosylated hemoglobin (HbA1c), C-reactive protein, tumor necrosis factor (TNF- α), transforming growth factor (TGF-ß1), and total antioxidant were studied. Ultrastructural and histopathological assessment of injury were also undertaken using transmission electron microscope. STZ-induced diabetes resulted in significant increase in the levels of blood glucose, HbA1c, C-reactive protein, TNF- α and TGF-ß1, and a decline in total antioxidant reserve of the myocardium. Administration of genistein to diabetic rats resulted in a decrease in blood glucose (p < 0.001), % HbA1c (p < 0.0001), C-reactive protein (p < 0.001), and expression of TNF- α (p < 0.001) and TGF-ß1 (p < 0.0001) proteins. In addition, genistein treatment results in augmentation of total antioxidant (p < 0.01) reserve of the hearts. The above findings were supported by histological as well as immunohistochemical localization of NF-κB (p65) in the heart. Genistein treatment ameliorated the ultrastructural degenerative changes in the cardiac tissues as compared to the diabetic control. The result demonstrates that genistein restored the integrity of the diabetic myocardium by virtue of its anti-inflammatory and antioxidant effects.

Survey of reasons for discontinuation from in vitro fertilization treatment among couples attending infertility clinic.

BACKGROUND: With the increase in infertility burden, more and more couples are opting for in vitro fertilization (IVF). Despite the availability of various treatment options, the major concern that needs to be addressed is the reasons why such couples, initially motivated so strongly, drop out in fairly high numbers from IVF cycles. With this point of view the study was designed. AIM: The objective of this study was to explore the reasons why couples discontinue fertility treatment. SETTINGS AND DESIGN: This retrospective study was carried out among couples in the age group of 20-40 years who opted for IVF at Tertiary care hospital and a private infertility center. MATERIALS AND METHODS: Medical records for 3 years (2009-2012) were taken out and included in the study for analysis. Socio-demographic details along with indication for IVF and reasons for drop-separate IVF therapy were recorded on case record form and were analyzed. RESULTS: Twenty-one percent of the patients had tubal pathology, thus making it the commonest female related factor for indication of IVF. Oligoasthenospermia (13%) was the commonest cause of male related infertility factor. Financial burden was the primary cause for terminating treatment in majority of the IVF cases. CONCLUSIONS: Financial burden (62.5%) was the commonest reason for drop out among couples from IVF cycle.

Bacopa monniera protects rat heart against ischaemia-reperfusion injury: role of key apoptotic regulatory proteins and enzymes.

OBJECTIVES: Rat isolated hearts were perfused in a Langendorff model to study the cardioprotective effects of Bacopa monniera, a medicinal herb used in the Indian system of medicine, on cardiomyocyte apoptosis and antioxidant status following ischaemia-reperfusion (I-R) injury. METHODS: Forty-eight rats were randomly divided into four groups (12 in each group): sham group (no ischaemia-reperfusion injury), B. monniera control group (orally fed B. monniera at a dose of 75 mg/kg, for three weeks); ischaemia-reperfusion control group(subjected to ischaemia-reperfusion-induced myocardial injury) and B. monniera-treated group (same protocol as ischaemia-reperfusion control group except that rats also fed B. monniera). KEY FINDINGS: Post-ischaemic reperfusion injury resulted in significant cardiac necrosis, apoptosis, depression of heart rate, decline in antioxidant status and elevation in lipid peroxidation. Oral administration of B. monniera per se for three weeks to healthy rats caused augmentation of myocardial antioxidants, superoxide dismutase, catalase and glutathione, along with induction of heat shock protein 72 (HSP72). Ischaemia-reperfusion-induced biochemical and histopathological perturbations were significantly prevented by B. monniera (75 mg/kg) pre-treatment. Interestingly, B. monniera also restored the antioxidant network of the myocardium and reduced myocardial apoptosis, caspase 3 and Bax protein expression. CONCLUSIONS: Histopathological studies and myocardial creatine phosphokinase content further confirmed the cardioprotective effects of B. monniera (75 mg/kg) in the experimental model of ischaemia-reperfusion injury. The study provides scientific basis for the putative therapeutic effect of B. monniera in ischaemic heart disease.

Withania somnifera provides cardioprotection and attenuates ischemia-reperfusion induced apoptosis.

BACKGROUND & AIMS: The present study was undertaken to evaluate the cardioprotective mechanisms of Withania somnifera (Ws), in the setting of ischemia and reperfusion (IR) injury. METHODS: Wistar rats were divided into three groups and received orally saline (sham, control IR) and Ws-50 mg/kg (Ws-IR), respectively, for 1 month. On the 31st day, in the rats of control IR and Ws-IR group, LAD coronary artery occlusion was undertaken for 45 min followed by 1 h reperfusion. Subsequently, all the animals were sacrificed for biochemical, immunohistochemical {Bax and Bcl-2 protein}, terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) positivity and histopathological studies. RESULTS: Post-ischemic reperfusion injury resulted in significant cardiac necrosis, apoptosis, decline in antioxidant status and elevation in lipid peroxidation in the IR control group as compared to sham. Ws prior-treatment favorably restored the myocardial oxidant-antioxidant balance, exerted marked anti-apoptotic effects {upregulated Bcl-2 (p<0.001) protein, decreased Bax (p<0.01) protein, and attenuated TUNEL positivity (p<0.01)}, and reduced myocardial damage as evidenced by histopathologic evaluation. CONCLUSIONS: The antioxidant and anti-apoptotic properties of Ws may contribute to the cardioprotective effects.