| Objectives1. Literature researchThrough the literature review, progress in the understanding of traditional Chinese medicine and Western Medicine on diabetic nephropathy, especially to understand the impact of blood stasis, inflammatory factors and related signal pathway on diabetic nephropathy and interventions, laying the theoretical basis for clinical research and experiment.2. Clinical StudyThrough clinical research, to understand the proportion of blood stasis in DN and the characteristics of blood stasis, clear effects on diabetic nephropathy; to investigate the correlation between TGF-β1, NF-κB and blood stasis; analysis of patients with blood stasis syndrome of blood stasis using, to provide basis for the research on the establishment and activating blood and resolving stasis Chinese medicine TCM preventing and treatment mechanism of DN.3. Experimental StudyTo explore the relationship between TGF-β1 and NF-κB pathway and renal damage through animal experiment, and give the Chinese herbal medicine composition of tanshinone A intervention, compared to observe the damage of kidney and improve the situation and the impact on TGF-β1 and NF-κ B pathway.In the cell experiments, cultured rat mesangial cells in vitro high glucose (HBZY-1), to observe the effects of high glucose on proliferation of rat mesangial cells, to observe and investigate the mechanism of intracellular TGF-β1 and NF-κB pathway traffic dialogue, further observe the effect of Huoxue traditional Chinese medicine composition of tanshinone ⅡA in rats renal mesangial cells and the intervention effect on TGF-β1 and NF-κB pathway dialogue, provide the basis for promoting blood circulation and removing blood stasis of Chinese medicine composition of tanshinone A control DN.Methods1. Literature researchSystematic literature review and comb in diabetic nephropathy of TCM disease name, etiology and pathogenesis, syndrome characteristics, treatment of traditional Chinese medicine and modern epidemiology, etiology and pathogenesis, modern treatment.2.Clinical studiesThrough a cross-sectional survey analysis of the clinical data of patients with DN in patients with blood stasis syndrome, blood stasis syndrome in understanding proportion in DN and the characteristics of. Prospective observation and comparison of blood stasis syndrome and non blood stasis syndrome of diabetic nephropathy patients during the course of the process of change of kidney damage degree, determine the effect of blood stasis in diabetic nephropathy. In addition, differences are observed in the comparison of DN of patients with blood stasis syndrome and non blood stasis syndrome in patients with serum TGF-β1 and NF-κB concentration, a clear correlation between these pathways and blood stasis, and through the questionnaire survey analysis of patients with blood stasis and Invigorate the circulation of drugs.3. Experimental StudyIn animal experiments, first by the intraperitoneal injection of STZ combined with high energy diet induced rat DN model, observation of the characterization of the model to observe the animal model, to observe the pathological changes in fully automatic biochemical instrument related indexes and HE, PAS staining, the expression of qRT-PCR, Western blot method for detection of renal TGF-β1 and NF-κB in order to investigate the relationship between TGF-β1 and NF-KB pathway and renal damage in DN rats; intramuscular injection of tanshinone Ⅱ A, compared to observe the damage of kidney and improve the situation and the impact on TGF-β1 and NF-κB pathway using the experiment method.In the cell experiments, in-vitro cultured rat mesangial cells (HBZY-1), the use of MTT technology to observe the effects of high glucose on proliferation of rat mesangial cells, and to explore the mechanisms of intracellular TGF-β1 and NF-κB pathway traffic dialog using siRNA technology, the use of MTT count, qRT-PCR to study the effect of Activating Blood Circulation Herbs composition tanshinone II A in rat renal mesangial cells and the intervention effect on TGF-β1 and NF-κB pathway dialogue.Results1.Literature researchLiterature research results show, diabetic nephropathy is an important complication of diabetes mellitus, with the rapid development and poor prognosis, etc. It has become a public health problem affecting human health. In the past to explore the pathogenesis, around the endowment deficiency, improper diet, emotional disorders and-excessive labor, the pathogenesis is deficiency of foundation the emergence of phlegm, stasis, turbidity, drugs and other pathological product, mutual cementation, produce sustained damage to the human body. The blood stasis is a key factor in the development and prognosis of DN occurred. Chinese medicine in the treatment of DN deficiency at the same time, often taken into account in the treatment of patients with blood stasis, phlegm damp, wet, etc. Modern research suggests, inflammatory response throughout the whole of diabetic nephropathy, TGF-β1 and NF-κB pathway dialogue affects the progress of the disease, treatment methods and drugs but still not exact.2.Clinical studiesClinical results show that:(1) DN-patients with blood stasis syndrome accounted for 74.2%, is the main type of syndrome, blood stasis syndrome is closely related to diabetic duration. DN of blood stasis in platelets, whole blood viscosity (high), whole blood viscosity (cut) (100s-1), whole blood viscosity (low shear) (50s-1) and blood sedimentation equation K value increased significantly, suggesting that blood stasis is the important pathological product of DN. (2)Compared with non blood stasis syndrome, blood stasis syndrome patients in unit time,24h urinary protein and GFR showed obvious dynamic change, TGF-β1, NF-κB appeared in serum significantly increased, suggesting that blood stasis is an important factor to promote the progression of DN, and is associated with inflammatory cytokines and related pathway activation. (3)The current drug laws promoting blood circulation and removing blood stasis analysis, using the frequency order of Radix Salviae Miltiorrhizae, Rhizoma Chuanxiong, motherwort, safflower, tree peony bark, rhubarb. Reflects the doctors used more promoting blood circulation drugs, less breaking blood drugs.3. Experimental StudyIn the DN rat models induced by STZ, depression, increase the amount of drinking water, urine volume increased, form thin and DN clinical symptoms, urinary protein and hyperglycemia, given tanshinone Ⅱ A intervention, the symptoms improved, urinary protein excretion and fasting blood glucose decreased, suggesting that the damage of kidney pathology reduce, may be associated with down-regulation of TGF-β 1 and NF-κB, up-regulation of Smad7.In high glucose induced rat mesangial cells along with the extension of time, cells were significantly increased,72h MTT counting results and more before the experiment have obvious difference. Give tanshinone Ⅱ A treatment, weaken, proliferation of mesangial cells and, tanshinone ⅡA high concentration (100 mol/L) with low concentration (10 mol/L) has more significant effect. In mesangial cells induced by high glucose added siTGF-beta 1, proliferation of mesangial cells decreased, decreased expression of NF-κB, while the Smad7 expression increased; join siP65 (important molecular NF-κB), proliferation of mesangial cells decreased, reducing the expression of TGF-β1,but Smad7 expression increased.Conclusion1. Literature researchDiabetic nephropathy is an important complication of diabetes mellitus, blood stasis is a key factor in the development and prognosis of DN occurred. Chinese medicine in the treatment of DN at the same time, promoting blood circulation to remove blood stasis runs through.2.Clinical researchBlood stasis is closely related to the course of diabetes. Blood stasis is not only an important pathological product of DN, it is also an important factor to promote progress in DN. Blood stasis and TGF-β1, the NF-κB pathway activation. In the treatment of DN blood stasis, promoting blood circulation drugs use more, broken blood drugs use fewer.3. Experimental StudyTanshinone ⅡA possibly through down-regulation of TGF-β1 and NF-κ B, up-regulation of Smad7 to improve DN kidney damage. In rat renal mesangial cell, there are interactive dialogue TGF-β1 and NF-κB pathway, Smad7 is one of the important nodes, tanshinone Ⅱ A can cut the TGF-β1 and NF-κB, raise of Smad7 inhibiting the proliferation of mesangial cells in high glucose. |
PDF Full Text Request