| Objective:Diabetic nephropathy(DN) is one of the most important microvascular complications of diabetes mellitus(DN), which is one of the most important causes of the end stage renal disease(ESRD). The pathogenesis of the DN is very complex,some studies found that persist inflammation is very important in the incident and development of DN. Recent studies have found that Janus Kinase(JAK)/ Signal transducers and activators of transcription(STAT) signaling pathway is required for the occurrence and development of DN. Recent studies have found that tripterygium wilfordii glycosides(TWP) and berberine(BBR) could decrease the level of proteinuria, but the mechanisms have not been sufficient elucidated. In our study, type2 diabetes mellitus(T2DM) rats as our object of study, and we used TWP and BBR as interventional medcine. The aim of this study is to determine the effect of the different doses of TWP and BBR on diabeties kidneys whether through inactivation the JAK/STAT signaling pathway.Methods:130 clean grade male SD rats were raised, 10 of which were randomly selected as normal control(NC) group, which were all fed with standard diet. Others were fed with high sugar and high fat diet for 8 weeks and streptozotocin(STZ)(30mg/kg)was injected through tail vein to induce T2 DM animal model. The successfully induced rats were randomly divided into eight groups: the diabetic rats without drug intervention(DM), TWP treatment group with low, middle and high doses(TL,1mg/kg·d; TM, 3mg/kg·d; TH, 6mg/kg·d), BBR group(150mg/kg·d), TWP combine with BBR(TBL, 1mg/kg·d+150mg/kg·d; TBM, 3mg/kg·d+150mg/kg·d; TBH,6mg/kg·d +150mg/kg·d). The SD rats were started with the TWP and BBR administration by gavage after the injuction of STZ. And the adminstration time is 8week. Among this study, blood and urine samples were collected for biochemical examination. The levels of ICAM-1 and IL-6 in serum were dectected by enzyme linked immunosorbent assay(ELISA). The m RNA levels of JAK2, STAT3, SOCS1,ICAM-1 and IL-6 were detected by q PCR, and protein levels of the p-JAK2,p-STAT3, SOCS1, ICAM-1 and IL-6 were detected by western blotting analysis. The distribution of ICAM-1 and IL-6 were detected by immunohistochemistry.Results1) Compared with the NC group, the level of TG, TC, FINS, HOMA-IR and HOMA-? were significantly increased in the DM group after the 8 weeks administration of high-sugar and high-fat diets(P<0.05), However the blood glucose have no significantly changes(P<0.05). Besides, the blood glucose was significantly increase after the injection of STZ compared with the NC group(P>0.05). After the administration of TWP and BBR, there was no difference in the level of blood glucose between DM group and the treatment of TWP and BBR(P>0.05), the level of ALT, AST, BUN, Scr and the number of the WBC and RBC have no significant change before and after the administration of TWP and BBR(P>0.05).2) The level of 24 h UMA: The level of UMA was higher in DM group than that in the NC group(P<0.05). However, after the administration of the TWP and BBR,the level of UMA was significantly decreased in a dose dependent(P<0.05).3) The levels of ICAM-1 and IL-6 in serum were dectected by ELISA:Compared with the NC group, the levels of ICAM-1 and IL-6 in serum were higher than those in the DM group(P<0.05). However, after the administration of the TWP and BBR, the levels of ICAM-1 and IL-6 were significantly decreased in a dose dependent(P<0.05).4) The level of JAK2?STAT3?SOCS1?ICAM-1 and IL-6 m RNA were detected by q PCR in kidney of SD rats: Compared with the NC group, the levels of SOCS1,ICAM-1 and IL-6 m RNA were higher than those in DM group(P<0.05). However,after the administration of the TWP and BBR, the levels of ICAM-1 and IL-6 m RNA were decreased and the level of SOCS1 was increased in a dose dependent(P<0.05).In the same time, we found the levels of JAK2 and STAT3 have no significant differences in each group(P>0.05).5) Western blot were applied to detect the changes of p-JAK2, p-STAT3, IL-6,ICAM-1 and SOCS1 at proteins levels in kidney: The levels of p-JAK2, p-STAT3,IL-6, ICAM-1 and SOCS1 were obviously increased in the renal cortex of the DM group compared with the NC group(P<0.05), and increased of p-JAK2, p-STAT3,ICAM-1 and IL-6 expressions were significantly inhibited by administration of TWP and BBR in a dose-dependent manner, however, the level of SOCS1 was obviously increased in a dose manner(P<0.05).6) The distribution of ICAM-1 and IL-6 were examined in glomerulus and tubules by immunohistochemical staining: The area and density of ICAM-1 and IL-6 weresignificantly increased in the glomeruli of DM rats compared with those in the NC group, positive particles were yellow brown, and renal tubular epithelial cells were also expressed partly(P<0.05). Compared with DM group, TWP and BBR decreased the distribution of IL-6 and ICAM-1 in the glomeruli and interstitium in a dose dependent(P<0.05).Conclusions:1. The activation of JAK/STAT signal pathway and the expression of ICAM-1 and IL-6 which could induce the injury of kidney in T2 DM rats.2. TWP and BBR could decrease the level of proteinuria in a dose dependent in T2 DM rats.3. TWP and BBR could decrease the expression of ICAM-1 and IL-6 of kidney in the T2 DM rats, may be related to inactivation the JAK2/STAT3 signaling pathway and increase the level of SOCS1. |
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