| Objective:To observe the expression of Cathepsin B and cystatin C in different stage of diabetic rats and investigates their potential roles,to study the change of CB,CC,ColIV and FN after treated by hirsutella sinensis, and to find the most possibly mechanism of this changeMethod:section one:Sixty rats were divided into diabetes mellitus group induced by intravenous injection of streptozotocin(55mg/kg) and normal group injected with citrate buffer.Ten rats sacrificed respectively at the end of fourth week,eighth week and sixteenth week in both groups.24h urine excretion was collected in each rat before sacrifice.The blood and the kidney were collected from each rat.The mRNA and protein expression of the CB and CC in kidney were detected by real time PCR and immunohistochemical staining respectively,section two:100 healthy male wistar rats were divided into three groups randomly as follows:normal rats Normal group),diabetic rats,the diabetic rat model was built by ntraperitoneal injection of STZ(55mg/kg),then the diabetic rats were divided randomly into model rats(model group) and treat rats(treat group).in the end of the fourth,eighth and sixteenth week,the urine of each rats was Collected,then ten rats of each groups were sacrificed and the kidney of them was harvested and blood were Collected.The expression of CB,CC,ColIV and FN in renal tissue were measured by real time PCR and imunohistochenical technique.Result:Compared with fourth week's,the expression of Ccr,24h urinary protein excretion,CB,CC of M group were increased significantly at the end of eighth week.(P＜0.01 or P＜0.05).with the aggravation of diabetic nephropathy,the expression of CC,ColIV,FN,24h urinary protein excretion were up-regulate significantly(P＜0.01 or P＜0.05).The expression of CB in M group was up-regulated at eighth week significantly(P＜0.01),whereas, at the end of sixteenth week it was down-regulated significantly (P＜0.01).The 24h urinary protein excretion and the protein and mRNA expression of ColIV,FN were negatively correlated with CB(P＜0.01).compare with Normal group,the serum glucose,renal weight,body weight,renal weight index of rats increased significantly(P＞0.05),but there were no significantly difference of serum glucose in model and treated groups(P＜0.01 or P＜0.05).compared with model group,the renal weight, body weight and renal weight index in treated group decreased obviously(P＜0.01),the Ccr and urine protein excretion of model group and treat group increased in the fourth week(P＜0.01),the expression of CB,CC, ColIV and FN was significantly up -regulated in eighth week(P＜0.01 or P＜0.05),but the expression of CB in model group in the sixteenth week was significantly down -regulated(P＜0.01).compare with the model group,the expression of CC,ColIV and FN in treat group were significantly downregulated in the sixteenth week(P＜0.01 or P＜0.05) but CB was up-regulated significantly(P＜0.01).Conclusions:The unbalance between CB and CC existed in diabetic nephropathy renal tissue which is likely to lead to the accumulation of ECM. After treated by hirsutella sinensis,the expression of CB were increased,on the contrary,the expression of CC was decreased obviously,the degrade of ColIV and FN were increasing.So the accumulations of ECM in diabetic rat's renal were decreased.So the most probably mechanism of hirsutella sinensis protect renal in diabetic rats is recover the balance of the expression of CB and CC.the protect effect of hirsutella sinensis to diabetic nephropathy renal is relative to the sustaining time of treatment. |
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