Intervention Effect Of Compatibility Of Salvianolic Acid A、B On PDGF-C/PDGFR-α Signaling Pathway In Renal Fibrosis

Objective: To investigate the intervention effect of Salvianolic acid A、B componentmolecules of drug compatibility on PDGF-C/PDGFR-α signalig pathway in renalfibrosis,which can establish the new mode of molecular medicine compatibility andprovide a new idea for clinical research of Chinese medicine to the molecular theory ofTCM medicine application.Methods:50male Sprague-Dawley rats were randomly divided into normal group (n=10), model group (n=40). The renal obstruction model of rats was established byunilateral ureteral obstruction (UUO).and then the model group were divided into thecontrol group (n=10), salvianolic acid A group (n=10), salvianolic acid B group (n=10)and salvianolic acid A and B group (n=10). Two days after the model were set up,corresponding treatments were given to individual group.Two weeks after treatment, allrats were killed for collecting samples. Renal histology， renal function, urinaryprotein NAG, urinary β2microglobulin, were examined and the expression of PDGF-C、PDGFR-α in rental tissue was tested by western bloting as well. Analysing the mRNAexpression levels of PDGF-C、PDGFR-α by RT-PCR technique.Results:1HE staining result In the normal group, the renal tissue no pathologicalchange; The renal tissue of model groups showed a large number of atrophic tubularepithelial cells, compensatory expansion and inflammatory cells infiltration.Renalinterstitial fibrosis was significantly.Compared with control group, the pathologicalchanges of kidney tissue were improved to different degree in all treatment groups. thedegree of pathological change in salvianolic acid B group and salvianolic acid A and Bgroup were significantly lower than that of salvianolic acid A group (P <0.05).2Masson staining result In the normal group,the renal tissue collagen deposition is not obvious, the model group showed renal capsule, tubular basement membranefilamentous apparent quality light blue collagen deposition, Compared with the modelgroup collagen deposition in each treatment group were significantly reduced. Eachtreatment group, salvianolic acid A and B group was significantly lower than salvianolicacid A group (P <0.05).3. Renal function test results：Compared with the normal group, model group and Salgroup B serum Cr levels were significantly increased (P <0.05)，Model group and thetreatment group, serum BUN levels were significantly increased (P <0.05). Comparedwith the model group, the treatment group showed decreases in serum Cr levels, but SalB group decreased obviously (P <0.05), Each treatment group showed decreases in serumBUN levels, but Sal group A and group B decreased Sal obviously (P <0.05), among thetreatment groups, serum Cr and BUN levels showed no significant difference (P>0.05).4. Urine β2microglobulin test results：Compared with the normal group, model groupand the treatment of urinary2-MG levels were significantly increased (P <0.05)，Compared with the model group, the treatment of urinary2-MG levels weresignificantly lower (P <0.05)，Between treatment groups, the difference was notstatistically significant (P>0.05).5. Urinary NAG protein test results：Compared with the normal group, model group andthe treatment of urinary NAG protein levels were significantly increased (P <0.05)，Compared with the model group, the treatment group were significantly lower urinaryNAG (P <0.05), Between treatment groups, the difference was not statisticallysignificant (P>0.05).6. CTGF immunofluorescence test results： CTGF expression signal was green,expression sites mainly localized in renal tubular epithelial cells. Normal rat kidneytubular epithelial cells were mostly negative, the fluorescence intensity is weak (±); Thenumber of rat renal tubular epithelial cells positive cells in model group than the controlgroup significantly increased the fluorescence intensity was significantly enhanced (++ +); By Sal A, B, and component compatibility after treatment, CTGF fluorescenceintensity was significantly reduced compared with the model group, in which Sal A+Bgroup decreased more significantly (±), followed by Sal group B (+), Sal group A slightdecrease compared with the model group fluorescence (++).7. Par-3immunofluorescence test results：Par-3expression signal was green, expressionsites mainly localized in the cytoplasm of renal tubular epithelial cells. Normal ratkidney tubular epithelial cells were mostly positive, strong fluorescence intensity (+++); The number of positive cells in rat renal tubular epithelial cells in the model groupwas significantly less than in the normal group, the fluorescence intensity wassignificantly reduced (±); By Sal A, B, and component compatibility after treatment, SalA, Sal B, Sal A+group B Par-3fluorescence intensity was significantly enhancedcompared with the model group, in which Sal A+B group enhanced obvious (+++),followed by Sal group B (++), Sal A group of slightly enhanced fluorescence (+).8.RT-PCR results：Compared with the normal group, the renal tissue of rats in the modelgroup PDGF-CmRNA and PDGFR-αmRNA relative expression was significantlyincreased (P <0.05); compared with the model group, the renal tissue of rats in eachtreatment group PDGF-CmRNA and PDGFR-αmRNA relative expression wassignificantly lower (P <0.05), among the treatment groups, Sal A+B group significantlydecreased renal tissue relative expression of PDGF-CmRNA, which is better than Salgroup A the difference was statistically significant (P <0.05), Sal A+B group cansignificantly reduce the renal tissue relative expression of PDGFR-αmRNA, which isbetter than Sal groups A and B, the difference was statistically significance (P <0.05).9.Western blot analysis：Compared with the normal group, the renal tissue PDGF-C andPDGFR-α protein in rat model group was significantly increased relative expression (P<0.05); compared with the model group, the renal tissue of rats in each treatment groupPDGF-C and PDGFR-α protein expression was significantly reduced relative (P <0.05),among the treatment groups, Sal A+B group of renal tissue relative expression ofPDGF-C protein was significantly lower in Sal group A and Sal group B (P <0.05), compared with Sal B group，the kidney tissue PDGFR-α protein relative expression ofSal group A+B has no significant difference (P>0.05).Conclusions:1. Sal B, A+B group can significantly improve the pathological changesin the kidney tissue, which is better than Sal Group A; Sal A+B group can significantlyreduce the deposition of collagen in rat kidney tissue, which is better than Dan phenol Sala+B group could significantly inhibit the expression of CTGF in renal tissue, which isbetter than Sal a and group B; acid a group Sal a+B group can significantly promotekidney tissue Par-3expression, which is better than Sal a and group B; Sal a+B groupcan significantly reduce the relative expression of PDGF-CmRNA kidney tissue, which isbetter than Sal group A; Sal a+B group can significantly reduce the relative expressionof PDGFR-αmRNA kidney tissue, which is better than Sal a and group B; Sal a+Bgroup can significantly reduce the relative expression in renal tissue PDGF-C protein, itseffect better than Sal A and group B; while Sal A, B and molecular medicine forcompatibility no significant difference in improvement in renal function and renal tubularfunction in rats with respect.2.Sal A, B and molecular medicine to some extent on the compatibility of improvingrenal function, renal tubular function and renal pathology, UUO kidney of rats has aprotective effect, which may be related to the promotion of Par-3in UUO large rat kidneytissue, reduced expression of CTGF, effectively reducing PDGF-C protein and geneexpression and PDGFR-α in the kidney tissue, renal fibrosis PDGF-C/PDGFR-αinterference signal pathway. Embodies the Chinese multi-channel, multi-target, thecombined effect of multiple ingredients.