| Objective:To investigate the effect of ASTRAGALI RADIX and SALVIAE MILTIORRHIZAE RADIX ET RHIZOMA Compatibility and their effective components drug serum on HGF and PAR-3expression levels and JAK/STAT signaling pathway of rat renal tubular epithelial cell under high glucose condition.Methods:To adopt the sixth generation NRK-52E cell which was divided into seven groups: high glucose group,fosinopril group, salvianolic acids group, astragalus saponins group, components combination of ASTRAGALI RADIX and SALVIAE MILTIORRHIZAE RADIX ET RHIZOMA1:1group, granular formulation combination1:1group,normal serum high glucose group.To observe the effect of drug serum on NRK-52E cell proliferation by MTT method.And test the purity of HGF and PAR-3in cell culture supertant by ELISA.Analysing the mRNA expression levels of HGF、PAR-3、JAK2、STAT1、STAT3by RT-PCR technique.Results:1MTT experiment Compared with high glucose group, ASTRAGALI RADIX and SALVIAE MILTIORRHIZAE RADIX ET RHIZOMA Compatibility and their effective components drug serum had no obvious effect on NRK-52E cell proliferation after24h and48h intervention, and there is no statistical significance.But after72h intervention,there had statistical significance that components combination of ASTRAGALI RADIX and SALVIAE MILTIORRHIZAE RADIX ET RHIZOMA1:1group could promote the cell proliferation (P<0.05).After96h intervention then, there had obvious statistical significance that drug serum groups promote the cell proliferation,particularly in fosinopril group, followed by components combination of ASTRAGALI RADIX and SALVIAE MILTIORRHIZAE RADIX ET RHIZOMA1:1group,salvianolic acids group, astragalus saponins group, granular formulation combinationl:1group (P<0.05).2ELISA Compared with high glucose group,there had no statistical significance of HGF expression levels in each group after four time points intervention. Fosinopril group, salvianolic acids group, astragalus saponins group could promote PAR-3expression levels after48h intervention (P<0.05).3.RT-PCR results: compared with high glucose group, there had obvious significance that Salvianolic acids group and granular formulation combination1:1group could promote HGF mRNA expression level after24h intervention (P<0.05).However,there had no significance of PAR-3mRNAexpression levels after24h intervention (P>0.05). ASTRAGALI RADIX and SALVIAE MILTIORRHIZAE RADIX ET RHIZOMA1:1group and fosinopril group could inhibit JAK2expression which had statistical significance (P<0.05).But STAT-1and STAT-3mRNA just had the decreasing tendency.There was no obvious significance in high glucose group and normal rat serum high glucose group (P>0.05)Conclusions:ASTRAGALI RADIX and SALVIAE MILTIORRHIZAE RADIX ET RHIZOMA Compatibility and their effective components drug serum had no poisonous effect on NRK-52E cell.The expression of HGF、PAR-3of renal tubular epithelial cell after drug serum intervention or inhibiting JAK2protein expression in JAK/STAT signaling pathway would exert its anti-fibrosis effect. Objective:To investigate the effect of different does groups of ASTRAGALI RADIX and SALVIAE MILTIORRHIZAE RADIX ET RHIZOMA’s effective components and their compatibility on Collagen deposition levels and JAK/STAT signaling pathway in UUO rats models.Methods:To adopt UUO kidney fibrosis rats models.The56SPF rats were randomly divided into7groups:normal group(n=8), model group(n=8), fosinopril group(n=10), components combination of ASTRAGALI RADIX and SALVIAE MILTIORRHIZAE RADIX ET RHIZOMA1:1group(n=10),components combination of ASTRAGALI RADIX and SALVIAE MILTIORRHIZAE RADIX ET RHIZOMA1:2group(n=10),components combination of ASTRAGALI RADIX and SALVIAE MILTIORRHIZAE RADIX ET RHIZOMA2:1group(n=10).Each group was treated with different does drugs after UUO surgery for14days. Exam serum creatinine (Scr) together with urea nitrogen(BUN) in blood, urinary β2-microglobulin (β2-MG) and retinol binding protein (RBP)by ELISA. To test JAK/STAT signaling pathway protein expression in UUO rats models by Western Blot.Results:1.Renal function The serum Cr level in model group is higher than normal group obviously (P<0.05). And fosinopril group Cr level is lower than model group obviously (P <0.05).There is no statiscal significance of residual groups’serum Cr level (P>0.05).Additionaly, There is no statiscal significance of all groups’serum BUN level (P>0.05);2.renal tubular protein examination Components combination of ASTRAGALI RADIX and SALVIAE MILTIORRHIZAE RADIX ET RHIZOMA1:1group and fosinopril group could decrease β2-MG、RBP level in rat urine (P<0.05), there is no statistical significance in residual groups (P>0.05).3. Tubulointerstitial pathological changes (1) HE staining There were not any pathological changes in normal kidney.UUO groups showed a large number of atrophic tubular epithelial cells (particularly the tubular in cortex and recent marrow) with vacuolar degeneration, tubular luminal expansion, and inflammatory cells infiltration.The pathological changes in the treatment groups improved in vary degrees.(2) Masson staining Deposition of collagen in renal tissue was not obvious.Light blue collagen could be seen in renal capsule, base material of nephric tubules, and mesenchyme in UUO groups.All the treatment groups showed less collagen deposition than the model group.4.Test results of JAK2, STAT1and STAT3in kidney by western blot technique.The component compatibility group, fosinopril group and granules compatibility of Astragalus and Salvia group Astragalus and its effective components can reduce the expression of JAK, STAT1, STAT3protein in renal tissue. Astragalus saponin group was not obvious.Conclusions:Components combination of ASTRAGALI RADIX and SALVIAE MILTIORRHIZAE RADIX ET RHIZOMA1:1group,components combination of ASTRAGALI RADIX and SALVIAE MILTIORRHIZAE RADIX ET RHIZOMA1:2group,components combination of ASTRAGALI RADIX and SALVIAE MILTIORRHIZAE RADIX ET RHIZOMA2:1group could decrease JAK2、STAT1、STAT3protein expression levels in kidney tissue,and there is no obvious relationship in different proportion groups. |
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