OBJECTIVE

Manihot esculenta (Crantz) leaves have been used for the management of diabetes based on cited ethnopharmacological studies. However, scientific evidence to support its efficacy is lacking. The aim of the study was to assess the cytotoxicity of the aqueous leaf extract and fractions of Manihot esculenta and its potential hypoglycemic effect on male Swiss albino mice.

The cytotoxicity assay was necessary to screen which extract and fractions will be used for the in vivo hypoglycemic study.

Phytochemical screening identified the composition of the aqueous crude extract and its fractions. The phytochemical results showed the presence of alkaloids, anthraquinone glycosides, carbohydrates, phenolic glycosides, saponins, and flavonoids.

Cytotoxicity was screened using CytoTox 96® (Promega), a NonRadioactive Cytotoxicity Assay on liver cancer (HepG2) and normal kidney (HK-2) cell lines. Five samples were tagged as highly cytotoxic and were flagged for further assays. These samples were DCM fraction (100 ppm), n-Hexane fraction (1 and 10 ppm), Ethyl acetate fraction (1 ppm), and Aqueous fraction (100 ppm).

The hypoglycemic activity was examined in alloxan-induced diabetic mice using in vivo hypoglycemic study. The aqueous crude extract at dose levels of 200 mg/kg and 300 mg/kg body weight showed significant reduction in blood glucose levels compared to the diabetic control but not exceeding the results in the metformin treatment group. A p-value of 0.05 set a priori was used to consider whether the intervention had a statistically significant difference compared to the diabetic control.

The effectiveness of aqueous crude extract in reduction of blood glucose in mice may be attributed to the synergistic effects of phytochemicals present, especially the alkaloids which were retained in the extract but were undetectable in the fractionated samples. The results of this study also support the findings of existing ethnopharmacological studies on M. esculenta leaf extract as a hypoglycemic agent.

Objective: Manihot esculenta (Crantz) leaves have been used for the management of diabetes based on cited ethnopharmacological studies. However, scientific evidence to support its efficacy is lacking. The aim of the study was to assess the cytotoxicity of the aqueous leaf extract and fractions of Manihot esculenta and its potential hypoglycemic effect on male Swiss albino mice. Methods: The cytotoxicity assay was necessary to screen which extract and fractions will be used for the in vivo hypoglycemic study. : Phytochemical screening identified the composition of the aqueous crude extract and its fractions. The phytochemical results showed the presence of alkaloids, anthraquinone glycosides, carbohydrates, phenolic glycosides, saponins, and flavonoids. : Cytotoxicity was screened using CytoTox 96® (Promega), a NonRadioactive Cytotoxicity Assay on liver cancer (HepG2) and normal kidney (HK-2) cell lines. Five samples were tagged as highly cytotoxic and were flagged for further assays. These samples were DCM fraction (100 ppm), n-Hexane fraction (1 and 10 ppm), Ethyl acetate fraction (1 ppm), and Aqueous fraction (100 ppm). Results: The hypoglycemic activity was examined in alloxan-induced diabetic mice using in vivo hypoglycemic study. The aqueous crude extract at dose levels of 200 mg/kg and 300 mg/kg body weight showed significant reduction in blood glucose levels compared to the diabetic control but not exceeding the results in the metformin treatment group. A p-value of 0.05 set a priori was used to consider whether the intervention had a statistically significant difference compared to the diabetic control. Conclusion The effectiveness of aqueous crude extract in reduction of blood glucose in mice may be attributed to the synergistic effects of phytochemicals present, especially the alkaloids which were retained in the extract but were undetectable in the fractionated samples. The results of this study also support the findings of existing ethnopharmacological studies on M. esculenta leaf extract as a hypoglycemic agent.
alkaloids ; hypoglycemic agents ; ethnopharmacology

Background: One of the therapeutic strategies for type 2 diabetes mellitus involves suppressing postprandial hyperglycemia by inhibiting key enzymes in carbohydrate digestion, α-glucosidase and α-amylase. While such inhibitors are commercially available, some researchers have turned to plants for potentially cheaper and safer alternatives. Objectives: The study aimed to investigate the in vitro α-glucosidase and α-amylase inhibitory activities of the leaf methanolic extracts of two native Philippine plants Ficus nota Blanco Merr. and Ficus septica Burm F, as well as their effects on postprandial blood glucose levels in a mouse model. Methodology: The in vitro activities of the leaf methanolic extracts were evaluated against porcine pancreatic α-amylase and yeast αglucosidase. The most active extract was partially purified into fractions by sequential solvent partitioning and subjected to in vitro testing. Postprandial antihyperglycemic activity was then assessed in normoglycemic ICR mice. Phytochemical analysis was also performed Results: The most active extract and fraction in vitro were FS-crude and FS-HexF, respectively, having significantly more potent αglucosidase inhibitory activity than the commercial drug acarbose. FS-crude and FS-HexF exhibited strong inhibition of αglucosidase and weak inhibition of α-amylase, which is considered favorable for novel inhibitors as it is hypothesized to reduce gastrointestinal adverse effects. However, FS-crude and FS-HexF did not significantly attenuate postprandial blood glucose levels in the oral starch tolerance test. Phytochemical analysis of FS-HexF putatively identified 6-gingerol as one of the possible bioactive components. Conclusion F. septica could be a potential source of glycoside inhibitors as it showed promising in vitro inhibition of α-amylase and α-glucosidase. While it did not exhibit significant postprandial antihyperglycemic activity in this study, more robust testing is recommended to make a definitive conclusion.
Amylases ; Glucosidases ; Hypoglycemic Agents

Humans ; Diabetes Mellitus, Type 1 ; Hypoglycemia/chemically induced* ; Hypoglycemic Agents/adverse effects* ; China ; Blood Glucose ; Insulin

OBJECTIVE: To observe the hypoglycemic effect of electroacupuncture (EA) at "Tianshu" (ST 25) combined with metformin on rats with type 2 diabetes mellitus (T2DM) as well as its effect on expression of adenosine monophosphate activated protein kinase (AMPK) in liver and pancreas. METHODS: Thirty-six male SD rats were randomly divided into a blank group (6 rats) and a model establishing group (30 rats). The rats in the model establishing group were fed with high-fat diet and treated with intraperitoneal injection of low-dose streptozotocin (STZ) to establish T2DM model. The rats with successful model establishment were randomly divided into a model group, a control group, a metformin group, an EA group and a combination group, 6 rats in each group. The rats in the EA group were treated with EA at "Tianshu" (ST 25), dense-disperse wave, 2 Hz/15 Hz in frequency and 2 mA in current intensity, 20 min each time. The rats in the metformin group were treated with intragastric administration of metformin (190 mg/kg) dissolved in 0.9% sodium chloride solution (2 mL/kg). The rats in the combination group were treated with EA at "Tianshu" (ST 25) and intragastric administration of metformin. The rats in the control group were treated with intragastric administration of 0.9% sodium chloride solution with the same dose. All the treatments were given once a day for 5 weeks. After the intervention, the body mass and random blood glucose were detected; the serum insulin level was detected by ELISA; the expression of AMPK and phosphorylated adenosine monophosphate activated protein kinase (p-AMPK) in liver and pancreas was detected by Western blot method; the expression of protein gene product 9.5 (PGP9.5) was detected by immunofluorescence. RESULTS: ①Compared with the blank group, the body mass in the model group was decreased (P<0.05); compared with the model group, the body mass in the EA group and the combination group was decreased (P<0.05); the body mass in the EA group and the combination group was lower than the metformin group (P<0.05). Compared with the blank group, the random blood glucose in the model group was increased (P<0.01); compared with the model group, the random blood glucose in the metformin group, the EA group and the combination group was decreased (P<0.01). The random blood glucose in the combination group was lower than the metformin group and the EA group (P<0.05). ②Compared with the blank group, the insulin level in the model group was decreased (P<0.05); compared with the model group, the insulin level in the metformin group, the EA group and the combination group was all increased (P<0.05). The insulin level in the combination group was higher than the metformin group and the EA group (P<0.05). ③Compared with the blank group, the protein expression of AMPK and p-AMPK in liver tissue was decreased (P<0.05), and the protein expression of AMPK and p-AMPK in pancreatic tissue was increased (P<0.05) in the model group. Compared with the model group, the protein expression of AMPK and p-AMPK in liver tissue in the metformin group, the EA group and the combination group was increased (P<0.05, P<0.01); the protein expression of AMPK in pancreatic tissue in the metformin group was increased (P<0.05); the protein expression of AMPK in pancreatic tissue in the EA group and the combination group was decreased (P<0.05); the protein expression of p-AMPK in pancreatic tissue in the metformin group, the EA group and the combination group was decreased (P<0.05). The protein expression of AMPK and p-AMPK in liver tissue in the combination group was higher than that in the metformin group and the EA group (P<0.05); the protein expression of AMPK in pancreatic tissue in the EA group and the combination group was less than that in the metformin group (P<0.05), and the expression of p-AMPK protein in pancreatic tissue in the combination group was less than that in the metformin group and the EA group (P<0.05). ④Compared with the blank group, the expression of PGP9.5 in pancreatic tissue in the model group was increased (P<0.01); compared with the model group, the expression of PGP9.5 in pancreatic tissue in the metformin group, the EA group and the combination group was decreased (P<0.05, P<0.01). The expression of PGP9.5 in pancreatic tissue in the EA group was lower than the metformin group and the combination group (P<0.05). CONCLUSION Electroacupuncture at "Tianshu" (ST 25) could promote the effect of metformin on activating AMPK in liver tissue of T2DM rats, improve the negative effect of metformin on AMPK in pancreatic tissue, and enhance the hypoglycemic effect of metformin. The mechanism may be related to the inhibition of pancreatic intrinsic nervous system.
Animals ; Male ; Rats ; Acupuncture Points ; AMP-Activated Protein Kinases/genetics* ; Blood Glucose ; Diabetes Mellitus, Type 2/drug therapy* ; Electroacupuncture ; Hypoglycemic Agents ; Insulins ; Metformin ; Rats, Sprague-Dawley

Humans ; Incretins/therapeutic use* ; Diabetes Mellitus, Type 2/drug therapy* ; Hypoglycemic Agents/therapeutic use* ; Risk Assessment

OBJECTIVE: The main aim of this study is to investigate whether acupuncture could be an effective complementary treatment for reducing the risk of macrovascular complications in diabetic patients currently taking antidiabetic medications using a nationwide population-based database. METHODS: We conducted a retrospective cohort study to assess the efficacy of acupuncture on cardiovascular complications in diabetic patients using data from patients between 40 and 79 years of age, newly diagnosed with diabetes between 2003 and 2006, found in the National Health Insurance Service-National Sample Cohort (NHIS-NSC) in Korea. From the data, we identified 21,232 diabetic patients who were taking antidiabetic medication between 2003 and 2006. The selected patients were divided into two groups-those who received acupuncture at least three times and those who received no acupuncture (non-acupuncture) in the year following their diagnosis of diabetes. After 1:1 propensity score matching (PSM), each group had 3350 patients, and the observation ceased at the occurrence of a major adverse cardiovascular event (MACE), which was defined as either myocardial infarction, stroke, or death due to cardiovascular cause. RESULTS: After PSM, the acupuncture group had a lower incidence of MACE (hazard ratio [HR]: 0.87; 95% confidence interval [CI]: 0.81-0.94; P = 0.0003) and all-cause mortality (HR: 0.77; 95% CI: 0.70-0.84; P < 0.0001) than the non-acupuncture group; the HRs for stroke-related mortality (HR: 0.75; 95% CI: 0.56-1.00; P = 0.0485), ischemic heart disease mortality (HR: 0.53; 95% CI: 0.34-0.84; P = 0.006) and circulatory system disease mortality (HR: 0.67; 95% CI: 0.55-0.82; P < 0.0001) were lower in the acupuncture group than in the non-acupuncture group in the secondary analysis. CONCLUSION Our results indicate that diabetic patients receiving acupuncture treatment might have a lower risk of MACE, all-cause mortality and cardiovascular mortality. This population-based retrospective study suggests beneficial effects of acupuncture in preventing macrovascular complications associated with diabetes. These findings call for further prospective cohort or experimental studies on acupuncture treatment for cardiovascular complications of diabetes. Please cite this article as: Jung H, Won T, Kim GY, Jang J, Yeo S, Lim S. Efficacy of acupuncture on cardiovascular complications in patients with diabetes mellitus in Korea: A nationwide retrospective cohort. J Integr Med. 2023; 21(2): 176-183.
Humans ; Retrospective Studies ; Cardiovascular Diseases/etiology* ; Diabetes Mellitus, Type 2/drug therapy* ; Hypoglycemic Agents/therapeutic use* ; Stroke/complications* ; Acupuncture Therapy ; Republic of Korea/epidemiology*

Metformin has been used for the treatment of type II diabetes mellitus for decades due to its safety, low cost, and outstanding hypoglycemic effect clinically. The mechanisms underlying these benefits are complex and still not fully understood. Inhibition of mitochondrial respiratory-chain complex I is the most described downstream mechanism of metformin, leading to reduced ATP production and activation of AMP-activated protein kinase (AMPK). Meanwhile, many novel targets of metformin have been gradually discovered. In recent years, multiple pre-clinical and clinical studies are committed to extend the indications of metformin in addition to diabetes. Herein, we summarized the benefits of metformin in four types of diseases, including metabolic associated diseases, cancer, aging and age-related diseases, neurological disorders. We comprehensively discussed the pharmacokinetic properties and the mechanisms of action, treatment strategies, the clinical application, the potential risk of metformin in various diseases. This review provides a brief summary of the benefits and concerns of metformin, aiming to interest scientists to consider and explore the common and specific mechanisms and guiding for the further research. Although there have been countless studies of metformin, longitudinal research in each field is still much warranted.
Humans ; Metformin/pharmacokinetics* ; Diabetes Mellitus, Type 2/metabolism* ; Hypoglycemic Agents/pharmacology* ; AMP-Activated Protein Kinases/metabolism* ; Aging

Diabetes mellitus is a chronic disease, typified by hyperglycemia resulting from failures in complex multifactorial metabolic functions, that requires life-long medication. Prolonged uncontrolled hyperglycemia leads to micro- and macro-vascular complications. Although antidiabetic drugs are prescribed as the first-line treatment, many of them lose efficacy over time or have severe side effects. There is a lack of in-depth study on the patents filed concerning the use of natural compounds to manage diabetes. Thus, this patent analysis provides a comprehensive report on the antidiabetic therapeutic activity of 6 phytocompounds when taken alone or in combinations. Four patent databases were searched, and 17,649 patents filed between 2001 and 2021 were retrieved. Of these, 139 patents for antidiabetic therapeutic aids that included berberine, curcumin, gingerol, gymnemic acid, gymnemagenin and mangiferin were analyzed. The results showed that these compounds alone or in combinations, targeting acetyl-coenzyme A carboxylase 2, serine/threonine protein kinase, α-amylase, α-glucosidase, lipooxygenase, phosphorylase, peroxisome proliferator-activated receptor-γ (PPARγ), protein tyrosine phosphatase 1B, PPARγ co-activator-1α, phosphoinositide 3-kinase and protein phosphatase 1 regulatory subunit 3C, could regulate glucose metabolism which are validated by pharmacological rationale. Synergism, or combination therapy, including different phytocompounds and plant extracts, has been studied extensively and found effective, whereas the efficacy of commercial drugs in combination with phytocompounds has not been studied in detail. Curcumin, gymnemic acid and mangiferin were found to be effective against diabetes-related complications. Please cite this article as: DasNandy A, Virge R, Hegde HV, Chattopadhyay D. A review of patent literature on the regulation of glucose metabolism by six phytocompounds in the management of diabetes mellitus and its complications. J Integr Med. 2023; 21(3): 226-235.
Humans ; PPAR gamma/metabolism* ; Curcumin/therapeutic use* ; Phosphatidylinositol 3-Kinases ; Diabetes Mellitus/drug therapy* ; Hypoglycemic Agents/pharmacology* ; Hyperglycemia/drug therapy* ; Glucose

OBJECTIVE: To explore the association between the use of metformin and the risk of ischemic stroke in patients with type 2 diabetes. METHODS: A prospective cohort study was designed from the Fangshan family cohort in Beijing. According to metformin use at baseline, 2 625 patients with type 2 diabetes in Fangshan, Beijing were divided into metformin group or non-metformin group and the incidence of ischemic stroke between the different groups during follow-up was estimated and compared by Cox proportional hazard regression model. The participants with metformin were first compared with all the parti-cipants who did not use metformin, and then were further compared with those who did not use hypoglycemic agents and those who used other hypoglycemic agents. RESULTS: The patients with type 2 diabetes were with an average age of (59.5±8.7) years, and 41.9% of them were male. The median follow-up time was 4.5 years. A total of 84 patients developed ischemic stroke during follow-up, with a crude incidence of 6.4 (95%CI: 5.0-7.7) per 1 000 person-years. Among all the participants, 1 149 (43.8%) took metformin, 1 476 (56.2%) were metformin non-users, including 593 (22.6%) used other hypoglycemic agents, and 883 (33.6%) did not use any hypoglycemic agents. Compared with metformin non-users, the Hazard ratio (HR) for ischemic stroke in metformin users was 0.58 (95%CI: 0.36-0.93; P = 0.024). Compared with other hypoglycemic agents, HR was 0.48 (95%CI: 0.28-0.84; P < 0.01); Compared with the group without hypoglycemic agents, HR was 0.65 (95%CI: 0.37-1.13; P=0.13). The association between metformin and ischemic stroke was statistically significant in the patients ≥ 60 years old compared with all the metformin non-users and those who used other hypoglycemic agents (HR: 0.48, 95%CI: 0.25-0.92; P < 0.05). Metformin use was associated with a lower incidence of ischemic stroke in the patients with good glycemic control (0.32, 95%CI: 0.13-0.77; P < 0.05). In the patients with poor glycemic control, and the association was not statistically significant (HR: 0.97, 95%CI: 0.53-1.79; P>0.05). There was an interaction between glycemic control and metformin use on incidence of ischemic stroke (P_interaction < 0.05). The results of the sensitivity analysis were consistent with the results in the main analysis. CONCLUSION Among patients with type 2 diabetic in rural areas of northern China, metformin use was associated with lower incidence of ischemic stroke, especially in patients older than 60 years. There was an interaction between glycemic control and metformin use in the incidence of ischemic stroke.
Humans ; Male ; Middle Aged ; Aged ; Female ; Metformin/adverse effects* ; Diabetes Mellitus, Type 2/drug therapy* ; Cohort Studies ; Ischemic Stroke/complications* ; Prospective Studies ; Hypoglycemic Agents/adverse effects* ; Stroke/prevention & control* ; Retrospective Studies