Experimental Study On Protective And Therapeutic Effect Of Shenshuning During The Development From Mesangial Proliferative Glomerulonephritis To Glomerulosclerosis In Rats

OBJECTIVE: Glomerulosclerqsis is the end-stage morphological expression of many kinds of renal diseases, and no effective way has been found to cure it currently. Mesangil proliferative glomerulonephritis (MsPGN) is one of the most common pathologic types of renal diseases, so it's very important to cure and prevent the development of MsPGN to FSGS. Based on the animal experiment, this research discussed the mechanism of glomerulosclerosis and the mechanism of Shenshuning(SSN) which preventing the development from MsPGN to glomerulonephritis.METHOD: The model of Mesangial Prolifereative Glomerulonephritis were made by injecting steophylocdccus enterotoxin B via rats' caudal vein, feeding bovine serum albumin, injecting Freunds adjuvant periodical. In order to make the glomerulonephritis rat model, the experiment period was prolonged to 16th and 20th week. The rats were treated with SSN, and benazepril was used as control. At the end of the 12th and 16th week, we examined the 24 hours urine protein quantity and the renal function , the renal morphological expression and ultrastructure, and deposits of circulating immune complexes via immunofluorescence. Immunohistochemical staining was performed to explore the extracellular matrix of the renal tissue as collegenIV(COLrV), fibronection(FN), a-smooth muscle actin (a-SMA), transforming growth factor β₁(TGF-β1) and basic fibroblast growth factor(bFGF). Reverse transcription-polymerase chain reaction (RT-PCR) was used to mensurate the gene expression of TGF-β1, bFGF, tissue inhibitor of metailoproteinase -1 (TIMP-1).RESULTS: The pathological evidence of 16th week indicates :extensive tubular atrophy, interstitial fibrosis, capillary loop appearing to be sclecrotic, mesangial and matrix proliferation subsiantialiy, capillary getting extensively deformed and some of it becoming partly obstructed under microscope. An increase in mesangial matrix and mesangial cells, effacement of foot processes locally, thickening of GBM, and irregular structure of GBM were found by means of Electron Microscopy. The Immunofluorescence shows IgG deposits in the mesangial and endothelial cells, which presents as shapes of granularity and clump, therefore, indicating that the MsPGN and FSGS have been obtainedsuccessfully.Rats of the MsPGN model group show more proteinuria, injury of renal function, pathological injury of mesangial cells, which worsened accompanied with sustained course of making model. While SSN and benazepril group show less injury compared with model group. The renal tissue morphologic showed the significantly protective effect to glomerulim capillary cavity and glomerula cells, the MsPGN model group rats show high expression of TGF-|31, bFGF, a -SMA,FN, the protein expression of Col IV at mesangial cells, and high mRNA expression of TGF- $ \, bFGF> TIMP-1 at the renal tissue. The two therapy groups both can decrease the expression of TGF- 3 i, bFGF, a -SMA,FN, the protein expression of ColIV at mesangial cells, and high mRNA expression of TGF-0 ]N bFGF^ TIMP-1 at the renal tissue. Compared with benazepril, the 16th week therapy effect of SSN appeared more significantly difference .The effects of SSN on MsPGN rats: SSN and benazepril can decreased proteinuria , protected renal function of the model rats. At 12th and 16th week, light microscopic show SSN can protect and repaired the glomeruii capillary network, renal cells, renal epithelia cells and interstitial tissule. The immunohistochemical staining reveal that the expression of collagen IV, FN is low in normal group. At 12& week, the protein expression of collagen IV and FN of MsPGN model rats are much higher than that of normal group (p<0.01). The protein expressions of collagen IV and FN of Glomerulosclerosis model rats at 16 weeks are much higher than that group at 12th week, it has evidently difference (p<0.01). At 12th and 16th week, SSN and benazepril treatment group rats' protein expression of collagen IV and FN are distinctly lower than model group. But for the same treatment group, there is no obvious difference between the result of 12th and 16th week. SSN show advantages than benazeprl at depressing FN. (p<0.05)The normal group rats show high expression of a-smooth muscle actin at capillary while not find at glomeruii and tubular. At I2th and 16th week, the MsPGN model group rats show high expression at the renal tissue, while the glomerulosclerosis model group rats are much higher (pO.Ol). At 12 and 16 week, the expression was obviously attenuated after treated with shenshuning and benazepril(P<0.01). For the same group, there is no obvious difference between the result of 12th and 16th week.