Clinical Study Of Diabetic Peripheral Neuropathy And Mechanism Tangbi Health Intervention

[Study background]Diabetic peripheral neuropathy (Diabetic peripheral neuropathy, DPN) is a common complication of diabetes mellitus (DM). DPN often involves sensory nerve and plant nerve, causing tingling, numbness, and even lead to exercise and nerve dysfunction in clinical. With high rate of incidence, it damages to health seriously and leads primary diabetes to disability. The world health organization estimates that there will be about360million DM patients in the world by2030, most of who suffer type2diabetes mellitus DPN. The pathogenesis of DPN is very complex and have not been fully elucidated causing limitation to the western medicine for the treatment and effectiveness yet. Traditional Chinese medicine treating DPN has overall advantages and obvious curative effect for its various means and ways. In recent years, there are many useful researches about DPN disease name, pathogenesis, clinical syndrome differentiation and treatment. The holistie therapy of DPN using TCM has its advantages and obviously curative effect. Professor Tonghua Liu insist the pathogenesis of DPN were deficiency of qi yin and poison stuck the shen collateral and brings up the new therapy of nourishing yin and qi, detoxify, dispersing blood stasis and clear the collateral. This project discusses the nerve protection mechanisms and targets of Chinese herbal compound Tangbikang treating DPN on the basis of previous research, and provides clinical basis of clinical application.[Objective]To objective evaluation of clinical efficacy and safety of treatment of diabetic peripheral neuropathy by Chinese herbal compound Tangbikang, the clinical trials were adopted by self control.[Methods]Clinic trial35DPN patients conforming to the diagnostic criteria, based on the basic treatment for blood pressure, blood glucose and blood lipid, were given Chinese herbal compound Tangbikang. After curing for8weeks, we measured some indicators, like TCM syndrome, laboratory examination and electromyography.Experimental research1.vivo experimentsThe experimental diabetes mellitus rats model were induced by feeding high fat forage and injection streptozotocin. Tangbikang groups were given Tangbikang with different dose, Body weight, blood glucose and sciatic nerves conduction velocitywere detected in every4weeks. Rats unilateral sciatic nerve were cutting-up after16weeks, pathological change of rat sciatic nerve of each group was observed with the light microscope, ultrastructure of rat sciatic nerve was observed by electron microscopy; the TC and TG were determined by the enzyme analysis method, the HDL-C and LDL-Cwere determined by immunoturbidimetry method; the p-EP,ET and FINS levels of rat plasma were determined by radioimmunoassay; the NEFA and NO levels of rat serum weredetermined by the method of Cul test paper; the expressions of NGF, BDNF, NT-3and VEGFproteinof rat sciatic nerve were determined by immunohistochemical assay and western blot; the expressions of NGF, BDNF, NT-3,and VEGF mRNA were detected by real-time fluorescence quantification.2.vitro experimentschwann cell model was establish in high glucose environment from schwann cells lineage.The effect of different dose Tangbikangon the proliferation of rat epidermal cells was detect by MTT method after stimulate by drug serum of different dose and serum at24h,48h,72h and96h. The content of TNF-a in supernatants were determined by radioimmunoassay after stimulate by drug serum of different dose and serum at48h; the content of sVCAM-1in supernatants were determined byELISA method; the expressions of schwann cell P38and P-P38were detected by Western blot method.[Results]Clinical trialAfter the treatment byTangbikang, the conduction velocitiesof MCv and SCvwas better than before, the difference was significant (P<0.05).Security Indexincludingblood glucose, glycosylated hemoglobin, blood lipid and blood pressurewith normal limits, the difference was insignificant (P>0.05), and there were no obvious side effects has been found during the treatment; the total effective rate of the Tangbikang was91.14%.Experimental research1.vivo experimentsCompared with normal group, model group weight was significantly reduced, the blood glucose was significantlyrisen, sciatic nerves conduction velocity were significantly reduced, the content of NO,β-EP, FINS,HDL-C, protein and mRNA of NGF, protein and mRNA of BDNF and protein and mRNA of NT-3were significantly risen; the content of ET, TG, TC、LDL-C、NEFA、protein and mRNA of VEGF were significantly reduced. Compared with model group, the rats body weight with different doses of Tangbikang were significantly increased; the content of NO,β-EP HDL-C,FINS, protein and mRNA of NGF, protein and mRNA of BDNF and protein and mRNA of NT-3were significantly reduced; the content of ET、TG、TC、LDL-C、NEFA、protein and mRNA of VEGF were significantly risen. The pathological results showed that sciatic nervefiber of normal group was hard-packed, homogeneous distribution and in alignment; and looselyarranged, larger gap and degeneration and fracture in multiple places of model group; degeneration and fracture of sciatic nerve was significantly reduced in Tangbikang group, it was hard-packed, homogeneous distribution and in alignment, but has degeneration and fracture insome places, the myelin sheath was thickened and even colouring. Electron microscopy (sem) results showed that the sciatic nervemyelin sheath was structural integrity, densification and well-distributed in normal group, the sciatic nervemyelin sheath was serious separation of lamellar structure, a disordered arrangement between each layer and a large number of vacuoles and gaps were in control group, the vacuoles and gaps of sciatic nerve were significantly reduced in Tangbikang group, but the lamellar structure was also abnormal, the structures of mitochondria and rough endoplasmic reticulum were in normality, structural integrityschwann cell was observed.2.vitro experimentCompared with blank group,50M GLU and75M GLU groups, the degree of cell proliferation was significantly reduced after24h,48h,72h and96h, but there were no significantly difference between50M GLU and75M GLU group. Compared with50mM model group, the degree of cell proliferation wassignificantly risenin Mecobalamin: Tangbikang1:1group, Mecobalamin:Tangbikang1:2group, Mecobalamin:Tangbikang1:8group.Compared with blank group,the contents of sVCAM-1, TNF-a. P38mitogen activated protein kinase and phosphorylated P38mitogen activated protein kinase were significantly risen, compared with50mM model group, the contents of sVCAM-1, TNF-a, P38mitogen activated protein kinase and phosphorylated P38mitogen activated protein kinase were significantly reduced in Mecobalamin:Tangbikanggroups.[Conclusion]Clinical trialChinese herbal compound Tangbikangcould improve of symptom, the nerve conductive velocity and the diabetic peripheral neuropathy integralsobviously, the total effective rate of the Tangbikang was91.14%.1.vivo experiments:1.1Chinese herbal compound Tangbikangcould improveblood glucose, body weight and blood lipid of STZ-DM rats; and reduce the content of non-esterified fatty acid in DPN rats serum; improvesciatic nerve conduction velocity of diabetic peripheral neuropathy rats, and it showed dose and time dependent.1.2Chinese herbal compound Tangbikang could protect the morphology ofsciatic nervein DPN rats, and repair its sciatic nerve myelin sheath and neuraxon.1.3Chinese herbal compound Tangbikang could rise the expression of FINS and NO in serum of DPN rats, reduce the expression of ET in plasma, these may be one of the mechanism of Tangbikang prevention and cure DPN.1.4Chinese herbal compound Tangbikang could rise the expression of P-EP in plasma of DPN rats, these may be one of the mechanism of Tangbikangneuroprotection and abirritation of DPN.1.5Chinese herbal compound Tangbikang could rise the expression of NGF, BDNF and NT-3protein and gene of sciatic nerve in DPN rats, reduce the expression of VEGF protein and gene of sciatic nerve in DPN rats, these may be one of the effective targets of Tangbikang prevention and cure DPN.2.vitro experiment2.1Tangbikang could proliferative activity of schwann cells in high sugar cultivation environment, Tangbikang is more effective then mecobalamin, Chinese herbal compound Tangbikangcould promoteproliferative activity of the rat sciatic nerve schwann cells,these may be one of the mechanism of Tangbikang neuroprotection and abirritation of DPN.2.2Tangbikang containing serum could reduce the expression of p38MAPK and activate the expression of p-p38MAPK protein of the rat sciatic nerve schwann cells in high sugar cultivation environment, and reduce the content of TNF-a protein in the rat sciatic nerve schwann cells supernatant, these may be one of the effective targets of Tangbikang neuroprotection and abirritation of DPN.2.3Tangbikang containing serum could reduce the content of sVCAM-1in the rat sciatic nerve schwann cells supernatant, this may be one of the effective targets of Tangbikang neuroprotection of DPN.