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 Wysłany: Sob 22:24, 05 Mar 2011 Temat postu: Cardiac function in type 2 diabetes and early coro |
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,[link widoczny dla zalogowanych]
Cardiac function in type 2 diabetes and early coronary flow reserve
The occurrence and development of diabetes plays an important role, the role of type 2 diabetes is inconclusive. In addition. Abnormal cell death in diabetic chronic vascular complications occurred well also play a role. We peripheral blood lymphocytes and pancreas of diabetic rats commanded owned small and Insulin on the death of two apoptosis-tung ring were studied. Diabetes to understand the culture of pancreatic cells in peripheral blood lymphocytes and the owner Insulin treatment of death as well as whether the inhibition of apoptosis. I. Materials and Methods 1. Animal model: selected male weighing 250 grams which Wistar rats were only 12 to l8 streptozotocin (ST'Z) 60mWkg [Fu in 01mmol / L sodium citric buffer 2 towels liquid malaria (pH4.5 .) STZ concentration of 1%] A Jung intraperitoneal injection. After 48 hours, blood glucose measured with ONETOUCHII type mouse tail vein blood,[link widoczny dla zalogowanych], blood culture barrier. When the blood culture ≥ 139mmoL / l time as diabetes; the remaining 6 rats were injected with the same volume of buffer solution of sodium lemon malaria. As a normal control group (N group). 12 Pui Pui diabetic rats were divided into diabetic group (D group) and Insulin treatment group (I group) 6, each free food and water. Group D to maintain blood glucose at a high level; I group according to the level of blood culture intraperitoneal injection of protamine zinc Insulin (PZI) 2 ~ 3U / d,[link widoczny dla zalogowanych], control any blood culture to 11ImmollL so. After 4 weeks the blood and pancreatic tissue to fill detection. 2 methods: (1) blood culture instrument with ONETOUCHII three groups of rats were measured before and after any venous blood of experimental training. (2) the blood from rat heart 3. Zan placed in sterile hepatic cord anticoagulation. The lymphocyte separation medium of the blood of the new unit: Add oo1l Shanghai Second Medical University Ninth People Hospital,[link widoczny dla zalogowanych], Prince Consort ■ Department of Geriatrics (Caiwen Wei, Zhu Jian), Department of Endocrinology (NG million blind}); Shanghai Second Medical University, Institute of immune (Shen Bai Hua, Lining Li) 7, /; isolated lymphocytes. After washing by PBS. Containing 10% human AB type serum RPMI1640 cell culture medium adjusted to 1.5 × 10/ral. Will be placed on inspection of the cell 37 ℃ 5% of the o. 2 for 24 hours. DNA ends by drawing cell labeling (TDT) tags. (3) of rat was removed under sterile pancreatic tail section (by Mallory ~ color stain in light microscopy confirmed the end of the ancient pancreatic islet cells more ...), placed in 2% newborn calf serum ancient (NSS) of saline in the cell suspension into 96 mesh nylon mesh filter, wash eulogy. The cells were transferred Qi 1 × 10,, in the DMEM containing 10% NBS medium. 37 ℃ 5% of the c02 for 24 hours,[link widoczny dla zalogowanych], to take out the Tunel staining. Observation of islet cell apoptosis. (4) Refer to Gorczyca and other mouth method. Slightly improved the cell pellet containing 1% formaldehyde PBS1ml, 4 ℃ 30 minutes. Washed twice. 0.5 TdT, 0.5nmol / 1 biological claim a 16-dUTP. 37 ℃ 3 hours after the removal. PBS after the addition of biological washing cable cyano fluorescent isothiocyanate labeled cable (FITc), at room temperature for 30 minutes. PBS wash. Death occurred a false charge of fine commanded DNA be broken. Labeled dUTP can be commanded in the flow meter and the fluorescence was fine shake fluorescence microscope was positive. (5) will be picking up the pancreatic tissue in 10% NBSRPMII640 placed into cell suspension. Add 0.5mg/mlRNAase digestion, 37 ℃ 30 分钟 propidium iodide (PI) staining and flow cytometry before the peak of computing Go/G1. Apoptosis in the amount of par value to the peak. (6) fluorescence microscope morphology of apoptosis: early apoptosis staining was ring; mid-and crescent-shaped patch of apoptosis was a small body; late apoptotic cell shrinkage, was a pull bright fluorescent group, or showed apoptotic bodies in the form of a dictionary of poly Village. 3. Statistical analysis: All the data are j ± funeral instructions. Difference between the groups using t test. Second, the results I change in blood glucose in three groups: Before and after the blood culture N group, respectively (2.764-0.3 mmol / L and (2.95 ± 0.36) mmol / l, D group, respectively (17.284-2.70) mmol / L and (16.5O4-305) mmol / l. Were significantly higher than the normal group (P <0001). I diabetic rat tissue culture Insulin treatment, blood glucose by the (1735 ± 29) mmol / L decreased (12424-182) mmd / l, blood sugar levels before and after treatment were significantly different (P <0.05) . Vertical ship 1
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