The standardized Jamu pahitan, an Indonesian antidiabetic formulation, stimulating the glucose uptake and insulin secretion in the in-vitro models.

Jamu pahitan is a polyherbal formulation commonly used for the traditional management of diabetes in Indonesia and is mainly prepared from Andrographis paniculata (Burm.f.) Nees. It is widely varied in herbal composition for every region has their own plant component addition to the formulation. A version of the formulation used in the greater Surakarta area contained five plant constituents. This study evaluated the in-vitro glucose uptake and insulin secretion stimulatory activities of Jamu pahitan to provide scientific evidence on its efficacy and safety of use. The water and ethanol extracts of three Jamu pahitan formulations were prepared. The total phenolic content (TPC) of the extracts was evaluated by the standard Folin-Ciocalteau method. Their effects on the viability of L6 skeletal muscle and RIN-m5F pancreatic cells were evaluated by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The glucose utilized by L6 myotubes treated with Jamu pahitan was assessed indirectly by the glucose oxidase method. The insulin secreted by RIN-m5F treated with the formulation extracts was analyzed by the enzyme-linked immunosorbent assay (ELISA). The correlation between TPC and the profile of safety and efficacy of the formulation was statistically evaluated. The water extracts of Jamu pahitan were safe and exerted significant glucose uptake and insulin secretion stimulatory activity in L6 and RIN-m5F, respectively. The ethanol extracts showed more potent effects than their water counterpart, albeit they exerted cytotoxic effects on the cells at the higher tested concentrations. The formulations at lower concentrations stimulated the proliferation of RIN-m5F. In addition, the TPC was strongly correlated with the glucose uptake and insulin secretion stimulatory activities and also the IC50 of the cells in positive manner. The present study supported the use of Jamu pahitan for the traditional management of diabetes in Indonesia by stimulating glucose uptake in the muscle cells and improving insulin secretion in ß-cells.

Hypoglycaemic activity of Mathurameha, a Thai traditional herbal formula aqueous extract, and its effect on biochemical profiles of streptozotocin-nicotinamide-induced diabetic rats.

BACKGROUND: The Thai traditional herbal formula-Mathurameha, consisting of 26 medicinal plants, has been used as an alternative and complementary medicine for diabetes treatment in Wangnamyen Hospital, Thailand. To provide scientific evidences on the efficacy and safety of this herbal formula, in vivo hypoglycaemic activity, effect on serum biochemical profiles and acute toxicity were investigated. METHODS: Experimental type 2 diabetes was induced in male Sprague-Dawley rats by intraperitoneal injection of nicotinamide 15 min prior to intravenous injection of streptozotocin. The most effective extract from the oral glucose tolerant test (OGTT) was administered daily via the oral route to diabetic rats for 2 weeks. Two-hour postprandial plasma glucose (2h-PPG) levels were measured on days 0, 7, and 14. Biochemical data were measured at the end of daily oral administration experiment. RESULTS: Aqueous extract of the herbal formula was the most potent extract for improving glucose tolerance of streptozotocin-nicotinamide-induced diabetic rats after single oral administration. After 2 weeks of daily oral administration, the aqueous extract showed a dose-dependent glucose lowering effect. At doses of 12.5, 25, and 50 mg/kg, the 2h-PPG level of diabetic rats decreased by 3.32%, 15.78%, and 17.94%, respectively. Most of the biochemical profiles of diabetic rats were improved, including the total cholesterol (TC), alkaline phosphatase (ALP), total protein, albumin, globulin, creatinine, and uric acid levels. The significantly increased triglyceride (TG) level observed in treated diabetic rats indicated a lack of a beneficial effect of the extract on lipid homeostasis. Nevertheless, there were no signs or symptoms of acute toxicity observed after oral administration of aqueous extract (5 g/kg) to both male and female rats. CONCLUSIONS: The results revealed that the herbal formula aqueous extract has hypoglycaemic activity, beneficial effects on biochemical profiles and a lack of acute toxicity. This study confirms the efficacy and safety of the Mathurameha herbal formula used for treating type 2 diabetes mellitus.

Development of phyllanthin-loaded self-microemulsifying drug delivery system for oral bioavailability enhancement.

Phyllanthin, a poorly water-soluble herbal active component from Phyllanthus amarus, exhibited a low oral bioavailability. This study aims at formulating self-microemulsifying drug delivery systems (SMEDDS) containing phyllanthin and evaluating their in-vitro and in-vivo performances. Excipient screening was carried out to select oil, surfactant and co-surfactant. Formulation development was based on pseudo-ternary phase diagrams and characteristics of resultant microemulsions. Influences of dilution, pH of media and phyllanthin content on droplet size of the resultant emulsions were studied. The optimized phyllanthin-loaded SMEDDS formulation (phy-SMEDDS) and the resultant microemulsions were characterized by viscosity, self-emulsification performance, stability, morphology, droplet size, polydispersity index and zeta potential. In-vitro dissolution and oral bioavailability in rats of phy-SMEDDS were studied and compared with those of plain phyllanthin. Phy-SMEDDS consisted of phyllanthin/Capryol 90/Cremophor RH 40/Transcutol P (1.38:39.45:44.38:14.79) in % w/w. Phy-SMEDDS could be emulsified completely within 6 min and formed fine microemulsions, with average droplet range of 27-42 nm. Phy-SMEDDS was robust to dilution and pH of dilution media while the resultant emulsion showed no phase separation or drug precipitation after 8 h dilution. The release of phyllanthin from phy-SMEDDS capsule was significantly faster than that of plain phyllanthin capsule irrespective of pH of dissolution media. Phy-SMEDDS was found to be stable for at least 6 months under accelerated condition. Oral absorption of phyllanthin in rats was significantly enhanced by SMEDDS as compared with plain phyllanthin. Our study indicated that SMEDDS for oral delivery of phyllanthin could be an option to enhance its bioavailability.

Antidiabetic Activities of Abutilon indicum (L.) Sweet Are Mediated by Enhancement of Adipocyte Differentiation and Activation of the GLUT1 Promoter.

Abutilon indicum (L.) Sweet is an Asian phytomedicine traditionally used to treat several disorders, including diabetes mellitus. However, molecular mechanisms supporting the antidiabetic effect of A. indicum L. remain unknown. The aim of this study was to evaluate whether extract of A. indicum L. improves insulin sensitivity. First, we observed the antidiabetic activity of aqueous extract of the entire plant (leaves, twigs and roots) of A. indicum L. on postprandial plasma glucose in diabetic rats. The subsequent experiments revealed that butanol fractions of the extract bind to PPARγ and activate 3T3-L1 differentiation. To measure glucose uptake enhanced by insulin-like activity, we used rat diaphragm incubated with various concentrations of the crude extract and found that the extract enhances glucose consumption in the incubated solution. Our data also indicate that the crude extract and the fractions (water and butanol) did not affect the activity of kinases involved in Akt and GSK-3ß pathways; however, the reporter assay showed that the crude extract could activate glucose transporter 1 (GLUT1) promoter activity. These results suggest that the extract from A. indicum L. may be beneficial for reducing insulin resistance through its potency in regulating adipocyte differentiation through PPARγ agonist activity, and increasing glucose utilization via GLUT1.

Antioxidant activity and antiapoptotic effect of Asparagus racemosus root extracts in human lung epithelial H460 cells.

The present study examined the antioxidant activity and protective effect of extracts from Asparagus racemosus roots against Lipofectamine-induced apoptosis. Five fractions from a successive extraction process ranging from non-polar to more polar solvents were obtained. The total saponin content as a marker in terms of diosgenin equivalent value of the root extracts was found to be in the range of 240-420 µg/mg extract, with higher values for the ethanol and aqueous fractions. The antioxidant activity measured using the DPPH method in terms of mean effective concentration (EC(50)) of the aqueous fraction was found to be 600 µg/ml as compared to 1.5 µg/ml of ascorbic acid. It is proposed that Asparagus racemosus root extracts effectively inhibit Lipofectamine-induced apoptosis by their protective effect, and may serve as an advantageous alternative option for gene delivery.

Molecular targets of apigenin in colorectal cancer cells: involvement of p21, NAG-1 and p53.

Persuasive epidemiological and experimental evidence suggests that dietary flavonoids have anti-cancer activity. Since conventional therapeutic and surgical approaches have not been able to fully control the incidence and outcome of most cancer types, including colorectal neoplasia, there is an urgent need to develop alternative approaches for the management of cancer. We sought to develop the best flavonoids for the inhibition of cell growth, and apigenin (flavone) proved to be the most promising compound in colorectal cancer cell growth arrest. Subsequently, we found that pro-apoptotic proteins (NAG-1 and p53) and cell cycle inhibitor (p21) were induced in the presence of apigenin, and kinase pathways, including PKCδ and ataxia telangiectasia mutated (ATM), play an important role in activating these proteins. The data generated by in vitro experiments were confirmed in an animal study using APC(MIN+) mice. Apigenin is able to reduce polyp numbers, accompanied by increasing p53 activation through phosphorylation in animal models. Our data suggest apparent beneficial effects of apigenin on colon cancer.

Nasal absorption and local tissue reaction of insulin nanocomplexes of trimethyl chitosan derivatives in rats.

OBJECTIVES: The objective of this work was to explore the potential and safety of trimethyl chitosan (TMC) and PEGylated TMC for improved absorption of insulin after nasal administration. METHODS: The nasal absorption of insulin nanocomplexes of TMC or PEGylated TMC was evaluated in anaesthetized rats. Concomitantly, the histopathological effects of these nanocomplexes on rat nasal mucosa were studied using a perfusion fixation technique. KEY FINDINGS: All insulin nanocomplexes containing TMC or PEGylated TMC showed a 34-47% reduction in the blood glucose concentration, when the insulin absorption through the rat nasal mucosa was measured indirectly. In addition, the relative pharmacodynamic bioavailability (F(dyn)) of the formulations was found to be dependent upon the charge ratio of insulin and polymer, regardless of polymer structure. The F(dyn) apparently decreased with increasing charge ratio of insulin : polymer. Although acute alterations in nasal morphology by the formulations were affected by the charge ratio of insulin and polymer, the formulation of insulin/PEGylated TMC nanocomplexes was shown to be less toxic to the nasal epithelial membrane than insulin/TMC nanocomplexes. CONCLUSIONS: PEGylated TMC nanocomplexes were a suitable absorption enhancer for nasal delivery of insulin.

Aqueous extract of Abutilon indicum Sweet inhibits glucose absorption and stimulates insulin secretion in rodents.

The objective of this study was to evaluate the antidiabetic effects of the aqueous extract derived from the Thai Abutilon indicum Sweet plant and to explore its effects on intestinal glucose absorption and insulin secretion. The authors hypothesized that the plasma glucose level could be reduced through the inhibition of glucose absorption and/or the enhancement of insulin secretion. Administration of the extract (0.5 and 1 g/kg body weight) in an oral glucose tolerance test led to a significant reduction in plasma glucose levels in 30 minutes after the administration in moderately diabetic rats, as compared with untreated rats (P < .05), and this was at a faster rate than the use of an antidiabetic drug, glibenclamide. The inhibition of glucose absorption through the small intestine was investigated using an everted intestinal sac. The results showed that the extract at concentrations of 0.156 to 5 mg/mL caused a reduction of glucose absorption in a dose response manner. The maximum response was noted at a dose of 2.5 mg/mL. The promotion of the extract on insulin secretion was confirmed by incubating beta cell of pancreatic islets and INS-1E insulinoma cells with the extract at 1 to 1000 microg/mL. These observations suggest that the aqueous extract from the A indicum plant has antidiabetic properties, which inhibited glucose absorption and stimulated insulin secretion. Phytochemical screening also revealed that the extract contained alkaloids, flavonoids, tannins, glycosides, and saponins that could account for the observed pharmacologic effects of the plant extract.

Development of mannosylated liposomes for bioadhesive oral drug delivery via M cells of Peyer's patches.

The aim of this study was to develop mannosylated liposomes as bioadhesive carriers for oral drug delivery. Two kinds of acyclovir (ACV)-entrapped mannosylated liposomes, i.e. ManN-ACV-lip and PAM-ACV-lip, were prepared by the use of mannosamine HCl (ManN) and p-aminophenyl-alpha-D-mannopyranoside (PAM), respectively. The mean sizes, drug entrapment efficiency, and loading capacity values of all liposomal formulations were in the ranges of 233-371 nm, 82-95%, and 42-47%, respectively. The mean size of PAM-ACV-lip was significantly smaller than those of conventional ACV liposomes and ManN-ACV-lip due to the more conical packing parameter of mannose-conjugated phospholipid. The mannosylating group grafted into bilayer membrane resulted in a decrease in drug entrapment, owing to competitive binding. The in vitro drug absorptions through everted sacs of mice ileum of both mannosylated ACV liposomes were significantly higher than those of conventional ACV liposomes or suspension.