Effect Of Polysaccharides From Marmeniorum On The Ischemia - Induced Diabetic Rats And Its Related Complications

Objective: To study Crepis polysaccharide(CTP) of alloxan diabetes model blood Ⅰglucose and insulin levels, the impact of glucose and lipid metabolism; research Crepis polysaccharide(CTP) of the heart and other related rat model diⅠ abetes the impact of complications, and to explore its mechanism.Method: Impact(1) Crepis polysaccharide(CTP) of alloxan type Ⅰdiabetic rats: intraperitoneal injection alloxan diabetic rats Ⅰtype; randomly divided into six groups, namely the control group( CON group); control group + Crepis polysaccharide(CON group + CTP group); model group(DM); model group + Crepis polysaccharide(DM + CTP group); model group with insulin(DM + insulin group) : model group + insulin + Crepis polysaccharide(DM group + insulin + CTP group) 10, male and female; Crepis polysaccharide(CTP) to 40 mg / kg dose oral administration; insulin group was given insulin subcutaneously, 3IU / only, insulin plus Crepis polysaccharide group was given the corresponding dose Crepis polysaccharide solution, while another subcutaneous injection 2IU / only insulin, blank add Crepis polysaccharide group received doses Crepis polysaccharide solution, blank group and model group was given normal saline, 1 / day, administered for 8 weeks. Measured once weekly body weight, blood glucose, food intake and water intake. Six weeks after the last administration before fasting 6h, 1h after oral administration 2g · kg-1 of glucose, glucose after detecting the change of administration in each group of rats after 0,30,60,120 min. Administered eight weeks after the last fasting 12 h, blood glucose 2h after administration of rat, retro-orbital venous plexus blood, serum preparation, serum insulin, cholesterol(TC), triglyceride(TG), high density lipoprotein(HDL), low density lipoprotein(LDL).(2)CTP alloxan-induced type Ⅰcardiac complications of diabetic rats rats: intraperitoneal injection of alloxan diabetic rat model Ⅰ; fasting after the last administration 12 h, intraperitoneal injection of 10% hydration chloral hydrate anesthesia, quickly made the carotid artery, connecting the ECG analyzer, measuring electrocardiogram; connection Bl-420 S biological and functional experimental measurement of mean arterial pressure, heart rate, pressure change rate and pressure decay rate. After rapid thoracotomy to expose the heart, blood, centrifugation, serum was prepared,-20 ℃preservation. For creatinine, creatine kinase(CK-MB), lactate dehydrogenase(LDH), C- reactive protein(CRP) content. Isolated rat heart, free excess tissue and weighed. And then separating the left and right ventricles, weighing, calculating cardiac hypertrophy index(CHI), left ventricular hypertrophy index(LV), left ventricle weight(LVW / RVW) than the right ventricle, the heart is heavier than the total weight(HW / BW); and measuring Left ventricular wall thickness(using micrometer screw left ventricular wall thickness); section ventricle with 4% paraformaldehyde-fixed, paraffin-embedded, cross-sectional cut the number of sheets every 1mm thick sections 4um to HE staining. Part of the heart tissue in a 3% glutaraldehyde and fixed for electron microscopy.Results:(1) stable Ⅰdiabetic rat model set up: with saline to alloxan(Alloxan) to a concentration of 5% solution, now with the current. Rats fasted 12 hours after a single intravenous injection of alloxan(160mg / kg) was added. 72 h after detection using random blood glucose meter(not fasting), blood glucose> 16.7mmol / L is made touch success, such as glucose <16.7mmol / L, again the tail vein injection of alloxan 100 mg / kg, periodic testing random blood glucose, glucose> 16.7mmol / L, and stable two weeks of successful modeling.(2) Crepis Crepis polysaccharide and polysaccharide joint subcutaneous insulin can induced weight gain T1 DM, compared with DM group, the DM + 5,8 week and a significant increase in body weight of rats INS CTP group INS group DM + + The difference was statistically significant, P <0.05 or P <0.01; compared with DM + INS group, DM + INS + CTP significantly decreased body weight in the first eight weeks, the difference was significant, P <0.05. Reduced food consumption, compared with DM group, the first 2,8 weeks DM + INS group and DM + INS + CTP food intake was significantly decreased, the difference was significant, P <0.05; compared with DM + INS group, DM + INS + CTP decreased food intake in the first eight weeks, the difference was not statistically significant. Reducing water intake, compared with DM group, the 2,5,8 weeks DM + INS group and DM + INS + water intake significantly decreased in rats CTP group, the difference was statistically significant, P <0.05 or P <0.01; and DM + INS group, DM + INS + CTP significantly decreased the amount of water in the first eight weeks, the difference was significant, P <0.05. Insulin levels did not change significantly. Oral glucose tolerance increase, compared with the DM group, DM INS significantly increased + INS group and DM + + CTP oral glucose tolerance group, the difference was significant, P <0.01; compared with DM + INS group, DM + INS + CTP group oral glucose tolerance increased, the difference was not statistically significant. Glycated hemoglobin decreased, compared with DM group, DM + INS group and DM + INS + CTP significantly reduce glycated hemoglobin levels, the difference was statistically significant P <0.05 or P <0.01.(3) Crepis polysaccharide can T1 DM rat serum lipid content decreased, compared with DM group, DM + INS group and DM + INS + CTP serum triglycerides and LDL were significantly reduced high-density lipoprotein protein significantly increased, P <0.01 or P <0.05; and DM + INS group, DM + INS + CTP group were significantly increased high-density lipoprotein levels, the difference was significant, P <0.05. Creatinine and urea nitrogen decreased, compared with DM group, DM + CTP group creatinine was significantly decreased, DM + INS group creatinine, blood urea nitrogen content was significantly reduced, P <0.05. Lactate dehydrogenase(LDH), creatine kinase isoenzyme(CK-MB) and C- reactive protein(CRP) levels were reduced, compared with DM group, DM + INS group and DM + INS + CTP serum LDH, CK-MB, CRP levels of significantly decreased, P <0.01 or P <0.05; compared with DM + INS group, DM + INS + CTP serum LDH, CK-MB, CRP levels of significantly decreased, P <0.05.(4) changes in the electrocardiogram of rats in each group: CON group and CON + ECG CTP group of rats were normal. DM rats were after abnormal electrocardiogram or myocardial infarction waveform. DM + INS + CTP group ECG abnormalities were less than other groups.(5) The kinetic parameters of each group blood flow in rats: Compared with CON group, blood pressure was significantly higher in DM group, the difference was statistically significant, P <0.05; compared with DM group, DM + INS group and DM + INS + CTP group reduced blood pressure, DM + INS + CTP group reduced blood pressure significantly, P <0.05; compared with DM + INS group, DM + INS + CTP group, blood pressure was significantly lower, the difference was significant, P <0.05.Compared with the CON group, DM group significantly increased heart rate, the difference was statistically significant, P <0.01; compared with DM group, DM + INS group and DM + INS + CTP group significantly decreased heart rate, the difference was significant, P <0.05; DM + INS + CTP group DM + INS group heart rate slightly decreased heart rate.Compared with the CON group, the pressure change rate DM group was significantly lower, the difference was significant, P <0.01; compared with DM group, DM + INS + CTP pressure change rate was significantly increased, the difference was statistically significant, P <0.05; DM + INS + CTP group pressure and heart rate change rate of DM + INS group were significantly increased, the difference was significant, P <0.05.Compared with the CON group, DM group pressure decay rate was significantly lower, the difference was significant, P <0.05; compared with DM group, DM + INS group pressure decay rate was significantly decreased, the difference was statistically significant, P <0.05; DM + INS + CTP group and heart rate DM + INS group pressure decay rate was significantly increased, the difference was statistically significant, P <0.05.(6) of the rats ventricular wall thickness changes: Compared with CON group, the rat heart CHI DM group, LVW / RVW and left ventricular wall thickness were significantly increased, the difference was statistically significant, P <0.05; and DM group, DM + INS group and DM + INS + CTP group heart CHI, LVW / RVW and left ventricular wall thickness were significantly lower, the difference was significant, P <0.05; DM + INS + CTP group than DM + INS group CHI significantly decreased, the difference was significant, P <0.05.(7) staining light microscopy test results myocardium of rats: CON group and CON + CTP group: no significant change.DM and DM + CTP group: cardiomyocytes extensive necrosis, severe muscle fiber rupture, a lot of muscle nuclei dissolving, disappearing, neutrophils large number of leaking.DM + INS group: most consecutive complete muscle fibers, edema is not obvious, morphologically normal nuclei. DM + INS + CTP Group: most consecutive complete muscle fibers, edema is not obvious, morphologically normal nuclei.(8) The results of electron microscopy to detect myocardial cells of rats: CON group and CON + CTP Group: visible myocardial cells intact, the muscle fibers arranged in neat, light and dark with clear, rich in mitochondria and normal structure ridge neat, dense; nuclear membrane integrity, chromatin uniform, fine.DM and DM + CTP groups: myofibrillar dissolution, myofascial mitochondrial membrane damaged significantly, showing mitochondrial swelling, cristae derangement, partial or total dissolved mitochondrial cristae and vacuoles, partial destruction of the nuclear membrane, and can see the withered dead bodies.DM + INS group: muscle fiber structure is substantially normal mitochondrial morphology were normal, but there are still some swelling of mitochondria, a small ridge disordered dissolution; muscle membrane, nuclear membrane intact.DM + INS + CTP group: the degree of structural damage of myocardial light, myofascial basic integrity of myofibrils arranged in parallel with the basic visible light and shade, and a few broken filaments; mitochondria accumulate, mild swelling, matrix fades, basic nuclear structure normal.Conclusion: CTP on blood sugar by intraperitoneal injection of alloxan-induced diabetic rat model Ⅰ, creatinine, urea nitrogen content was not significantly changed.CTP can be improved by intraperitoneal injection of alloxan-induced diabetic rats Ⅰtype of lipid metabolism in vivo.CTP joint subcutaneous injection of insulin can reduce type 1 diabetic rat serum CRP, CK-MB and LDH levels and protect cardiomyocytes from injury, but also can improve hemodynamics, inhibiting cardiac hypertrophy, myocardial fibrosis.