Dpn Rat Ar. The Ari Treatment, Parpmrna Expression And Oxidative Stress Changes In Research

Objective To study a process of insulin resistance and having a pathological process similar to human type 2 diabetic peripheral neuropathy in animal model approach, features, experimental value, and provide a reliable experimental evidence for the clinical research of type 2 diabetes mellitus complicated by chronic Patients with diabetic peripheral neuropathy (DPN) in the pathogenesis and mechanism of drug therapy.Methods 8 weeks old weighing 180-220g SD male rats 120 were randomly divided into 30 control group and 90 in the model. Control rats with normal diet. In the model rats fed with high fat high sugar diet for 4 weeks, the first pulse intraperitoneal injection of streptozotocin (STZ,35mg/kg), detection of random blood glucose levels. 4 weeks after the second intraperitoneal injection (STZ,45mg/kg), detection of random blood glucose levels, glycated hemoglobin enzymatic colorimetric method. Automatic biochemical analyzer lipid water Level, measured by radioimmunoassay fasting insulin and insulin sensitivity index (lnsulinsensitivity index, IS I). Random blood glucose≥16.7mmol/L were to become the standard model, and models were prepared in 4 weeks and 8 weeks after the measurement of body weight and blood sugar levels, where the blood glucose level<15 mmol/L rats to be removed. Then the normal physiological nerve function as the baseline value, at 4 weeks after STZ injection and 8 weeks to diabetic rats in the nervous system into a model neural electrophysiological function tests, the use of sensory nerve conduction velocity, motor nerve conduction velocity, H reflex latency, M wave amplitude changes measured by the sciatic nerve, tibial nerve can be excited about change of circumstances to determine whether there is peripheral neuropathy in diabetic rats.Results The diabetic rat model of peripheral neuropathy elevated blood lipids:LDL was 0.75±0.32mmol/L, TC was 1.43±1.07mmol/L, TG 1.72±0.6mmol/L, compared with the control group was statistically significant (P<0.05); reduced insulin sensitivity, sensitivity index:-2.61±0.34, compared with the control group was statistically significant (P<0.05); glycosylated hemoglobin increased:GHb 6.71 ±1.37%, with the control group statistical significance compared (P<0.05); modeling process physiological tibial nerve:0.66ms, compared with the control group was±H wave latency:10.49±0.66ms, statistically significant (P<0.01); M wave amplitude 4.20±2.96mv, compared with the control group was statistically significant (P<0.01); SNCV38.44±1.56m/s, compared with the control group was statistically significant (P<0.01); MNCV37.36±1.14m/s, compared with the control group was statistically significant (P<0.01).Conclusion:1. high sugar high fat diet fed with two different doses of STZ injection can be successfully copied with type 2 diabetes similar to human pathogenic mechanisms of peripheral neuropathy in type 2 diabetic rats with experimental peripheral neuropathy model, can be used for type 2 diabetes mellitus complicated by chronic Patients with diabetic peripheral neuropathy (DPN) pathogenesis and mechanism of drug therapy to be investigated further.2. DPN model of hyperglycemia, hyperlipidemia, insulin resistance, neuro-physiological changes, such as:nerve EMG, sensory nerve conduction velocity, motor nerve conduction velocity and changes in peripheral neuropathy of type 2 diabetes characteristics, is the study of type 2 pathogenesis of diabetic neuropathy and peripheral lesions of the ideal animal model for drug research. Objective To study the AR, PARPmRNA expression of peripheral nerve and blood in DPN rats and AR, oxidative stress, PARP activity of changes in the role in the pathogenesis of DPN and clear ARI (epalrestat) before and after intervention in DPN AR, Oxidative stress, PARP changes associated with the pathogenesis of DPN, and ARI (epalrestat) protective mechanism of the treatment, and ARI treatment of the role of targets and key mechanism for the clinical application of ARI treatment of DPN to provide a more reliable experimental evidence.Methods Experimental rat model using DPN, DPN in the STZ induction into the model after 8 weeks (12 weeks of experiment) the experimental rats were divided into 4 groups:nDM(ARI-); nDM(ARI+); DPN(ARI-); DPN(ARI+). Application of ARI (epalrestat) 50mg·kg-1·day-1 orally means dissolved administered intervention for 6 weeks, Rats were sacrificed 19 weeks after the experimental, and detected sciatic nerve AR, PARP molecule mRNA expression in injured nerves, tissue distribution and cellular localization of expression by FISH, and rat femoral artery experiments AR, PARP for relative quantification Expression by RT-PCR method, and ultrastructural changes of experimental rat sciatic nerve by, compared Epalrestat changes before and after treatmet.Results 1, FISH analysis showed that:The nDM(ARI-) and nDM(ARI+), ARmR NA,PARPmRNA expression of the myelin Schwann cells, nerve membrane, myelin, the axon is only within a few and specificity After the probe made w-ith the green fluorescent signal in the fluorescence microscope. It showed that. the ex-pression of AR mRNA, PARP mRNA in normal peripheral nerve tissue without the DM have only a very small amount of ARmRNA, PARPmRNA g-ene expression, without the DM application of ARI(epalrestat) treatment to nor-mal peripheral nerve tissue ARmRNA, PARPmRNA have no significant chang-es in gene expression.The DPN(ARI-), the myelin Schwann cells, nerve memb-rane, myelin, the axon shows a large number of the AR mRNA, PARP mRNA combined with the specific probe made of green fluorescence signal that whent he damagednerve DPN The large number of AR mRNA, PARPmRNA express-ion in the fluorescence microscope., suggesting that AR activation and oxidati-ve stress, PARP activation in the pathogenesis of DPN play an important role. The DPN(ARI+), the myelin Schwann cells, nerve membrane, myelin, the axon shows a small amount of the AR mRNA, PARPmRNA combined with the sp-ecific probe madeof green fluorescence signal in the fluorescence microscope. suggesting that theARI epalrestat can reduce DPN peripheral nerve tissue AR mRNA, PARP mRNA expression,and reduce the AR, PARP activity, and oxida-tive stress.2 The transmission electron microscope sciatic nerve-tibial nerve ultrastructur e:The nDM(ARF) and nDM(ARI+) sciatic nerve myelin integrity and smooth, full, distortion-free variant form, while the DPN(ARF) group showed the sciatic nerve-tibial nerve myelin twisted variant, delamination, swelling, dissolution, loss; axonal shrinkage, necrosis; mitochondria and endoplasmic reticulum swell-ing;schwann cell nuclei pyknosis, swelling, and debris formation.3,l),ARmRNA 2-ΔΔct, variance analysis:RT-PCR 2-ΔΔct relative quantification:T-he nDM(ARI-), nDM(ARI+), DPN(ARI-), DPN(ARI+) 4 groups ARmRNA quan-titative F=134.182, p=0.000, indicating p<0.05, significantly different, statistical-ly significant. The DPN(ARI-)and nDM(ARI-) and nDM(ARI+), the DPN(ARI+) compared, p<0.01, significantly different, statistically significant. The DPN(ARI+) and DPN(ARI-), the nDM(ARI-)and nDM(ARI+), compared, p<0.01, significan-tly different, statistically significant. The nDM(ARI-) ARmRNA with the nDM (ARI+), compared, p> 0.05, the difference was not statistically significant. The DPN(ARI-)and the DPN(ARI+) compared, p<0.01,significantly different, statisti-ccally significant.2),PARPmRNA 2-ΔΔct, variance analysis:RT-PCR 2-ΔΔct relative quantification: nDM(ARI-), the nDM(ARI+), the DPN(ARI-), the DPN(ARI+) 4 groups PARP mRNA 2-ΔΔct, quantitative F=578.051, p=0.000, indicating p<0.05, significantly different, statistically significant. The DPN(ARI-) and nDM(ARI-) and nDM(AR I+), the DPN(ARI+) compared, p<0.01, significantly different, statistically signi- ficant. The DPN(ARI+) and DPN(ARI-), the nDM(ARI-)nd nDM(ARI+), comp-areed, p<0.01, significantly different, statistically significant. The nDM(ARI-)and the nDM(ARI+),compared, p>0.05, the difference was not statistically significant. The DPN(ARI-) and the DPN(ARI+) compared, p<0.01, significantly different, statistically significant.Conclusion 1,DPN rats with peripheral nerve tissue and blood AR, PARP mRN A expression increased, AR, PARP activity increased, suggesting that AR, PAR P activation and oxidative stress play an important role in the pathogenesis of DPN.AR, PARP activation and oxidative stress was caused by a series of perip heralnerve injury DPN metabolism, pathology, pathophysiology involved in the process of cooccurrence of DPN, development, Showing that there exists close intrinsic links among metabolic abnormalities of DM-AR activation-oxidative stress-PARPactivation-vascular and nerve lesions-DPN.2, ARI (Epalrestat) can reduce the peripheral nerve tissue and blood AR, PAR P mRNA expression, to reduce the AR, PARP activity, and oxidative stress.And reduce the AR, PARP activity and the role of AR, PARPactivation by reducing AR, PARP mRNA expression of peripheral nerve tissue and blood, which cau-sed increased oxidative stress, DNA damage, metabolic abnormalities, pathology injury,and played against oxidative stress, improving metabolic injurys of DPN.3, The results clarified the role of ARI site and AR activity and oxidative str-ess and the relationship among PARP activity and further clarify the mechanic-smof and ARI treatment of DPN key factors, suggesting regularity of ARI tre-atment of diabetic animals, providing reliable experimental evidence for clinical application of ARI treatment of DPN.