Study On The Mechanism Of Tongluo Baoshen Recipe On Renal Interstitial Fibrosis In UUO Rat Model

1Objective:In order to elucidate the mechanism of Compound Tongluobaoshen(TLBS) Prescription preventing and curing renal interstitial fibrosis, delay Renal pathological changes, we observed the effectivity of TLBS Solution by established the obstructed kidney after unilateral ureteral obstruction (UUO) model.We also researched the effection of TLBS Solution to the cytokine (TGF-?1?CTGF) and ECM(FN. Col I ColIII) from levels of organ, tissue, protein and gene.2Method:Male SD55rats were randomly divided according to weights. Rats weight was measured weekly, and24h Urine protein, Scr?BUN and NAG were measured at three weeks. Then3weekend take the blood of rats tests biochemical indicator.The kidney was taken at3weekend to be observed the changes of the pathomorphology by using HE and Mallory staining method The cytokine (TGF-?1? CTGF) and ECM(FN. Col I?ColIII) protein was detected in rats renal tissue by immunohistochemical method. The cytokine (TGF-?1? CTGF) mRNA was detected in rats renal tissue by In Real-Time PCR method.3Results3.1Weight of the rats gradually increased,two weeks after modeling, TLBS group compared with the model group the body weight of rats increased significantly(P<0.05);three weeks after the modeling, the benazepril group and TLBS compound group weight gain, but no significant difference compared with model group.3.2Model group, the renal hypertrophy index (kidney weight/body weight) increased significantly, compared to the treatment groups Renal hypertrophy index decreased (p <0.05).3.3Model rats24h urinary protein increased, no significant difference between the treatment groups.3.4Model rats of serum Scr increased significantly, compared with, reduced to varying degrees in all treatment groups (P<0.05or P<0.01); model rats,BUN and NAG increased,but the treatment group increasedobvi-Ous.3.5Kidney histopathology observed:normal group and sham operation group, no pathological changes of the kidneys, and inflammatory cell infiltration increased in model group, tubulointerstitial, and renal tubular epithelial cell vacuolar degeneration, necrosis, tubular dilatation, atrophy, or even the structure disappearance and the interstitial space was widened, a large number of collagen fibril formation, the emergence of severe renal interstitial fibrosis.Benetton Group and TLBS Group of interstitial inflammatory cell infiltration, tubular epithelial cell degeneration, atrophy and interstitial fibrosis is lighter than the UUO model group.3.6Immunohistochemical results:cell factor(TGF-?1in CTGF) and extracellular matrix(FN, Coll, Col-Ill collagen)in the normal group, sham operation group,only a small amount of expression,increased expression of the model group and treatment group.compared with model group,bena-zepril group and TLBS group,expression were significantly reduced.3.7Real-time PCR results:cell factor (TGF-?1in CTGF) mRNA in the normal group, sham operation group, only a small amount of expression, increased expression of the model group and treatment group, compared with model group, benazepril group and TLBS group,expression were significant-ly reduced.4. Conclusion4.1Pharmacodynamic results show:the TLBS compound group can increase the UUO rats weight; reduced renal hypertrophy index; lower serum Scr, BUN, and NAG; alleviate the renal pathological changes in UUO rats4.2The TLBS compound treatment of renal interstitial fibrosis (RIF) mechanism: the protein level, The TLBS compound could be reduced UUO rats cytokines (TGF-?1in CTGF) and extracellular matrix (FN? col I, Col. III) protein expression, and thus delay the development of the RIF. The gene level, the TLBS compound could be reduced UUO rats cell factor1(TGF-beta1, CTGF) mRNA expression, and thus delay the development of the RIF.In summary, through a variety of experimental methods of evidence, TLBS compound have a positive effect in rats RIF. Its prevention and treatment of RIF mechanism is complicated.