Wednesday, March 07, 2007 10:52:49 AM
RESEARCH DESIGN AND METHODS
This study was conducted in the Department of Human Nutrition, NWFP Agricultural University, Peshawar, Pakistan and was approved by the Ethics Committee and Human Studies Review Board of the University of Peshawar. Selection criteria for the study included the following for people with type 2 diabetes: age >40 years, not on insulin therapy, not taking medicine for other health conditions, and fasting blood glucose levels between 7.8 and 22.2 mmol/l (140–400 mg/dl). A total of 60 individuals with type 2 diabetes, 30 men and 30 women, were selected for the study. The mean age of the subjects was 52.0 ± 6.87 years in the placebo groups and 52.0 ± 5.85 years in the groups consuming cinnamon. The duration of diabetes was also similar: 6.73 ± 2.32 years for the placebo group and 7.10 ± 3.29 years for the cinnamon groups. There was also an equal number of men and women in the placebo and cinnamon groups. All subjects were taking sulfonylurea drugs, i.e., glibenclamide; medications did not change during the study.
Cinnamon (Cinnamomum cassia) certified by the Office of the Director, Research and Development/Non-Timber Forest Products, NWFP Forest Department, Peshawar, Pakistan, was used in this study. Cinnamon and wheat flour were ground finely and put into capsules (Mehran Traders Pharmaceutical Suppliers, Peshawar, Pakistan). Each capsule contained either 500 mg of cinnamon or wheat flour. Both the cinnamon and placebo capsules were packaged in plastic bags containing 40 capsules (1 g or two capsules per day for 20 days), 120 capsules (3 g or six capsules per day for 20 days), or 240 capsules (6 g or 12 capsules per day for 20 days) and prepared for distribution to the subjects. When subjects finished testing after the first 20 days, they were given the second package of capsules. Compliance was monitored by capsule count and contact with the subjects. Compliance was considered excellent and all capsules were consumed.
The study was conducted for 60 days with 60 individuals with type 2 diabetes divided randomly into six equal groups. Group 1 consumed two 500-mg capsules of cinnamon per day, group 2 consumed six capsules of cinnamon per day, and group 3 consumed 12 capsules of cinnamon per day. Groups 4, 5, and 6 were assigned to respective placebo groups, which consumed a corresponding number of capsules containing wheat flour. Subjects consumed their normal diets and continued their medications throughout the study. From days 41 to 60, no cinnamon or placebo was given. The 1-g dose of cinnamon and placebo was spread over the day as 0.5 g (one capsule) after lunch and 0.5 g after dinner. The 3-g and 6-g doses of cinnamon and placebo were spread over the day as 1 g (two capsules) and 2 g (four capsules) after breakfast, lunch, and dinner, respectively. The subjects were instructed to take the capsules immediately after meals.
On days 0, 20, 40, and 60, 5 ml of fasting blood was collected from each subject. Blood samples were transferred to sterilized centrifuge tubes and allowed to clot at room temperature. The blood samples were centrifuged for 10 min in a tabletop clinical centrifuge at 4,000 rpm for serum separation. Serum samples were stored in a freezer at 0°C for later analyses. Glucose level was determined using an autoanalyzer (Express Plus; Ciba Corning Diagnostics, Palo Alto, CA). Triglyceride levels were determined by the enzymatic colorimetric method of Werner et al. (http://care.diabetesjournals.org/cgi/content/full/26/12/3215#R13#R13 13) using an autoanalyzer (Express Plus; Ciba Corning) and an Elitech kit (Meditek Instrument, Peshawar, Pakistan). Cholesterol levels were determined by enzymatic colorimetric method of Allain et al. (http://care.diabetesjournals.org/cgi/content/full/26/12/3215#R14#R14 14) using the same autoanalyzer. Chylomicrons, VLDL, and LDL were precipitated by adding phosphotungstic acid and magnesium ions to the sample. Centrifugation left only the HDL in the supernatant (http://care.diabetesjournals.org/cgi/content/full/26/12/3215#R15#R15 15). LDL cholesterol was calculated by dividing the triglycerides by 5 and subtracting the HDL cholesterol (http://care.diabetesjournals.org/cgi/content/full/26/12/3215#R16#R16 16).
Two-way ANOVA and randomized complete block design were used for statistical analysis (http://care.diabetesjournals.org/cgi/content/full/26/12/3215#R17#R17 17). Values are means ± SD.
Human Clinical Study: Healthy Volunteers – Russian scientist Dr. M. Abidoff evaluated the Glucose lowering properties of blueberry leaves extract in double blind placebo controlled study at Moscow Center for Modern Medicine, Russian Ministry for National Defense Industries (Abidoff 1999, Abidoff-Farma), Russia.
According to this research the CA and HCA makes up as much as 18±2% of blueberry leaves extract. Seventy-five healthy volunteers age between 37-66 years were invited to participate in double -blind placebo-controlled five-week trial. Sixty days before beginning the drug phase of this trial, volunteers underwent a period of diet counseling and surveillance. Their dietary intakes were standardized to contain 55-60% % total calories from carbohydrates. At 3-week intervals throughout the study, volunteers were evaluated for fasting plasma glucose values. Standard food record analysis and a symptom questionnaire were also included at the laboratory intervention times.
After the initial dietary run-in phase, subjects were randomly assigned to receive 150mg of standardized blueberry leaves extract standardized or placebo, to be taken three times a day in 200 ml of water before meals. Results of this study revealed that there was significant change in blood glucose values for both groups of volunteers. The after meal blood glucose level increased from 102mg/dL±8md/dl (baseline) to 142mg/dL ±7mg/dL in placebo group. Those volunteers taking the blueberry leaves extract plasma glucose level increase from approximately 109mg/dL ± 9 mg /dL to 121mg/dL±6mg/dL. The results of this clinical trial indicate that the blueberry leaves extract possess physiologically significant glucose lowering property. According to Prof. Abidoff MD (1999) the glucose-lowering effect of blueberry leaves extract is due to unique properties of chlorogenic acid to inhibit the activity of G6P key enzyme in glycogenolysis and gluconeogenesis, although the direct inhibition of intestinal amylase– key enzyme in dietary carbohydrates absorption, by the blueberry leaves extract cannot be ruled out. In addition, results of Welsh et al. (1987) indicated that chlorogenic acid could inhibit the intestinal absorption of glucose.
Human Clinical Study: Diabetes Patients – In a second clinical trial the effect of blueberry leaves extract on plasma glucose level was studied in patients with Type II Diabetics (Abidoff 1999) Twenty-nine patients average age 50 years with type II diabetes were selected to participate in double -blind placebo-controlled 60 days trial. Sixty days before beginning the drug phase of clinical study, patients underwent a period of diet counseling and surveillance. Their dietary intakes were standardized to contain 40-45% total calories from carbohydrates. Patients in the study were asked to maintain their medications throughout the dietary and drug phase of the trial. On admission and at two-week intervals throughout the study, patients were evaluated for fasting glucose, triglycerides serum values. Food record analysis, body mass index, and a symptom questionnaire were also included at the laboratory intervention times. After this initial dietary run-in phase, subjects were randomly assigned to receive 200mg of standardized blueberry leaves extract powder in capsule
form or placebo, to be taken three times a day in 200 ml of water before meals. During the initial period of diet counseling there was no significant change in fasting blood glucose values for either of the groups. However, beginning with week 6 and continuing to the end of the trial, those individuals taking the blueberry leaves extract showed a
significant reduction in mean plasma glucose levels, from approximately 169 mg/dL to 136 mg/dL (p < 0.01). Furthermore, by the end of the clinical study, those taking the blueberry leaves extract showed a reduction triglyceride and LDL values from 179 ± 95 mg/dL to 130 ± 53 mg/L (p < 0.005) and 141 ± 47 mg/dL to 115 ± 34 mg/dL (p < 0.01) respectively. All patients tolerated well blueberry leaves extract at even 400mg/ three times a day (1200mg/day). Results of the clinical trial are confirmed well known and previously described phenomenon that the blueberry leaves extract possess antidiabetic properties. The use of blueberry leaves extract may provide a first line approach to the reduction of blood glucose in type II diabetes patients before other prescriptive avenues are employed. Improvement in total cholesterol and LDL level observed in various studies is possible due to protective role of caffeic and chlorogenic acids in LDL oxidation that was recently described in scientific literature.
Blueberry leaves extract: Science Re-discovers Another Ancient Truth of Folk Medicine. The long history of Blueberry leaves extract use in folk medicine and its growing popularity by the informed public is no longer a scientific mystery. Blueberry leaves extract is a safe, natural, potent source of critical chlorogenic and caffeic acids and has a long and venerable history and an even more promising future in the long-term care of diabetics everywhere. Once again, as has been the case with many other leading health supplements for the last two decades, science finally, reluctantly proves the efficacy of what it once not only ignored but openly denigrated! Yet what is profoundly fascinating is how often and how accurately the lore and legends of folk medicine so often proves to clinically accurate! When we delve deeper into the fast-growing, impressive body of this new research, we find in many cases the active compound(s) identified and responsible for an herb or food's traditional health benefits have been somehow empirically understood and used for the appropriate purpose in some form by many of our ancestors well into the past. Often such use occurred for thousands of years, in a variety of different cultures, spread over many continents. Although the folklore may have been local, taken as a whole the use of these natural medicines were regional, if not actually global. Ironically, modern researchers appear to be learning once again about the complex relationships between our bodies and these traditional food and medicine plants in a somewhat backward fashion. If researchers took a more humble and open-minded approach to the potential wisdom of traditional medicines like Blueberry leaves extract, they might accelerate their understanding and serve the mass acceptance and use of these very real "cultural treasures". Genuine scientific enthusiasm for this task would help set better standards for product formulation, consumer education, and public regulation, which, in turn, could help alleviate and even prevent the suffering of millions of people around the world from major chronic diseases such as diabetes in the future.
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