| Diabetes is one of the chronic diseases doing great harm to human health, and diabetic nephrophathy is the main cause of End Stage Renal Disease (ESRD). At present, two problems exist in the establishment of diabetic nephropathy model. Firstly, the development of kidney damage is fairly slow. Secondly, because of the high-does usage of STZ, insulin had to be given to rats in order to control metabolic disturbance and keep the hyperglycemia on a middle or high level. Based on the above reasons, we have established the animal model of diabetic nephropathy and observed rat models' general condition, blood pressure, blood sugar, urinary protein in 24hours, renal function and pathologic abnormalities.Study shows that hypersugar can produce albuminuria. The typical clinical features of diabetic nephropathy include hypertension and albuminuria. At the same time, the extent of these two features positively relates to patients' prognosis. Effective control of albuminuria and hypertension can relieve the course of diabetic nephropathy worsening to renal functional failure. As our traditional medicine, Chinese medicine is very promising in treatment of immuno-inflammatory diseases. It has already been known that Bailin capsule is isolated from aweto. Its main contents include D-mannite, cordycepic acid, carrier alkaloid, 19 Kinds of amino acid, some vitamins and microelements. Bailin can protect renal tubular Na+-K+ATP enzyme, motivate the production of EGF (epidermal growth factor) by renal tubular epithelial cells, improve immunologic competence and stimulate renal tubular epithelial cells' proliferation. All of these can partially explain its effects, including restraining renal tubular atrophy and renal interstitial fibrosis. It is not quite cleur how Bailin Capsule acts on diabetic nephropathy so far. Based on the successful establishment of animal model, this study gave a further insight into the protective effect of Bailin capsule on diabetic nephropathy in rats.Recent research has indicated that progressive fibrosis features largely in diabetic nephropathy. It has been found that growth factor and cytokine play an important role in the generation and development of diabetic nephropathy Transforming growth factor-β1(TGF-β1) and connective tissue growth factor (CTGF) are both indispensable during the pathologic process. Studies also show that some herbal medicine could delay the progression of diabetic disease by affecting TGF-β1 and CTGF. In our study, the results show Bailin capsule, a Chinese herbal medicine preparation, can reduce the blood sugar level of rat diabetic nephropathy model, the blood pressure and albuminuria/24h and renal damage and meliorate renal function. This study focused on Bailin capsule's impact on the expression of rat models' TGF-β1 and CTGF, trying to elucidate diabetic nephropathy mechanism.Part one Experimental study of rats with diabetic nephropathyObjective To establish model of diabetic nephropathic rats, and to observe their general condition, blood pressure, blood sugar, urinary protein in 24hours, renal function, pathological changes of renal tissue.Methods Thirty-two rats were randomly divided into control group (control group), diabetic nephropathic model group (model group), which had two sub-groups according to different time (4w, 12w), every group had 8 rats. We established the rat model of diabetic nephropathy induced by streptozotocin (STZ), observed the blood sugar, urinary protein in 24hours, serum creatinine (Scr), blood urea nitrogen (BUN), and counted endogenous creatinine clearance rate (Ccr); we also observed the renal pathological changes by optics microscope and electronic microscope.Results The blood sugar of model group rats which were injected STZ was higher than that of control group in 2w, 4w, 6w, 8w, and 12w. Average arterial pressure, urinary protein in 24hours, SCr, BUN of model group rats were higher distinctly than those of control group, while Ccr of model group rats was lower than that of control group (P<0.05 for all). Furthermore, we found that the indexs of glomerulus sclerosis and renal tubular injury in the model group were also higher (P<0.05 for both).Conclusion We established successfully the rat model of diabetic nephropathy, which was the basis of our futher research of drug intervention.Part two Experimental study on effect of Bailin capsule on diabetic nephropathy in ratsExperiment one: Effect of Bailin capsule on biochemic al indexs and renal histopathological changes in rats with diabetic nephropathyObjective To observe the effect of Bailin capsule on blood pressure, blood sugar, urinary protein in 24hours, renal function, and pathological changes of renal tissue in rats with diabetic nephropathy.Methods Twenty-four rats were randomly grouped into control, model and treatment, 8 rats for each group. Two days after models' establishment, Bailin capsule was administrated (2.5g/kg) by a stomach tube in treatment group, and same amount of water was fed in model and control group. Then we detected the blood sugar, urinary protein in 24hours, Scr, BUN, calculated Ccr, and used optics microscope and electronic microscope to observe the the renal pathological changes.Results After 12w, the blood sugar, average arterial pressure, urinary protein in 24hours, Scr and BUN in model group were significantly higher compared with control group (P<0.05 for all). However, Ccr in model group was lower than that of control group, and there was obvious renal tissue damage (P<().05). After treatment of Bailin capsule, the average arterial pressure, urinary protein/24h, Scr, BUN were reduced, Ccr increased and renal damage eased (P<0.05 for all). Conclusion Bailin treatment can protect diabetic nephropathy rat models from kidney damage.Experiment two: Effect of Bailin capsule on the expression of TGF-β1 and CTGF in renal tissue of diabetic nephropathy rat modelObjective To observe the effect of Bailin capsule on the expression of transforming growth factor-β1 (TGF-β1) and connective tissue growth factor (CTGF) in rats with diabetic nephropathy.Methods Twenty-four rats were randomly grouped as control, model and treatment. There were 8 rats for each group. The expressions of TGF-β1 and CTGF in renal tissue were investigated by RT-PCR and immuno-histochemiccl method.Results In the control group, they had low level or nil expression of TGF-β1 and CTGF. In the model group, in contrast, expression of TGF-β1 and CTGF significantly increased (P<0.05 for both). After treatment of Bailin capsule, expression of TGF-β1 and CTGF was obviously reduced (P<0.05 for both). Correlation analysis showed that the expression of TGF-β1 and CTGF was positively relating to the damage of renal tissue. Their expression correlated with the indexs of glomerulus sclerosis and renal tubular injury (P<0.05 for both).Conclusion Bailin capsule can reduce the expression of TGF-β1 and CTGF in rats with diabetic nephropathy, which can reduce renal tissue injury. |
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