| Objective:To investigate the expression and the function of RAGE in pancreas and kidney of rats with SAP, and study the influence and the mechanism of QYG on the expression of RAGE in pancreas and kidney of rats with SAP.Methods:Eighty SD(Sprague-Dawley) rats were randomly divided into four groups:①SAP group (n=24),②QYG treatment group (n=24),③sham-operation(SO) group (n=24),④normal control group(n=8),8rats were killed in12h,24h and48h respectively in the former three groups; SAP model of rats were induced by injecting5%sodium taurocholate (STC)(0.1ml/100g) though retrograde cholangiopancreatography, rats in SO group were injected saline (0.1ml/100g) instead. All rats were injected saline (lml/100g weight) on the outside of the legs after modeling successfully. Rats in second group were given QYG (lml/100g) through gavage after modeling for two hours, and then administered once every12hours, the QYG was insteaded by saline in first group and third. The ratio of wet weight and dry weight of kidney and the weight coefficient of kidney were measured before and after treatment. The level of amylase (AMS), Malondialdehyde (MDA) and Superoxide dismutase (SOD) in serum were tested by colorimetriy. The concentration TNF-alpha, IL-2and IL-6in serum were measured by Enzyme-linked immunosorbent assay (ELISA). The expression of RAGE protein and it’s mRNA in pancreas and kidney were detected by Immunohistochemistry, Western blotting and RT-PCR methodes.Results:(1) The phenomenon of lots of inflammatory cells infiltrated, tissue congested and edema, hemorrhage or necrosis was obseraved in pancreas and kidney in first and second groups, The pathology score of pancreas and kidney in second group was lower than that in first group after treatment of QYG (P<0.05); There were no significant difference of pathological changes in pancreas and kidney in the latter two groups.(2) The ratio of wet weight and dry weight of kidney in the former two groups were lower than that the latter two groups (P<0.05), the ratio of wet and dry weight of kidney in the second group was increased after treatment of QYG, there was no statistical significant difference of the ratios between the second group and the third group at48h.(3) The coefficient of kidney in the first group was smaller than the latter three groups; the difference between each groups were not statistically significant (P>0.05).(4) The leveles of AMS and MDA in serum of the former two groups were both higher than the third group, which were decreased significantly in the second group after QYG treatment (P<0.05), which showed a gradual downward trend in three time point of12h,24h and48h. The total serum SOD levels in the former two groups were lower than that in third group, which were elevated in the second group after QYG treatment, but no significant difference was observed within12h, however which were increased significantly at24h (P<0.05).(5) The contents of IL-6and TNF-alpha in serum were higher in the former two groups than that in the third group, which were rised more abviously in the second group than that in the first group, and peaked at24h. But the levels of IL-2in serum showed an opposite performance, the efficacy of QYG shew a time-dependent relationship. IL-2and TNF-alpha showed a negative correlation (r=-0.873), IL-6and TNF-alpha showed a negative correlation (r=-0.897), IL-6and TNF-alpha exhibitted a positive correlation (r=0.931).(6) There were numbers of RAGE expression positive cells in tissues of pancreas and kidney in the former two groups, which were positively correlated with the severity of lesions. the expression leveles of RAGE in pancreas and kidney were reduced in the second group after QYG treatment when compared with that in the first group. The results of the RAGE protein and it’s RNA contents change trend tested by Western blotting and RT-PCR had the similar trend with Immunohistochemistry.(7) The efficacy of QYG for SAP was in a time-dependent manner.(8) After the statistical correlation analysis, the expression quantitation of RAGE protein in tissues of pancreas and kidney of rats showed a positive correlation with serum TNF-alpha contents(r=0.865, P<0.05), and which showed a positive correlation with serum IL-6contents(r=0.879, P<0.05), and a negative relationship with serum IL-2levels(r=-0.794, P<0.05). Conclusions:(1) The level of AMS in serum of rats in former two groups, which indicated the models of SAP were established successfully.(2) There were lots of RAGE proteins expression in pancreas and kidney of rats with SAP, RAGE may be involved in the damage of pancreas and kidney in rats with SAP.(3) Oxidative stress have took part in the process of SAP, the ability of antioxygen of the body was depressed by SAP.(4) The RAGE protein and it’s RNA in pancreas and kidney were down-regulated by QYG treatment, which can lessen the injury of pancreas and kidney of rats with SAP.(5)The protective mechnism of QYG in the damages of pancreas and kidney in rats with SAP, which maybe work by inhibiting the inflammatory response, reducing the production of AGEs, and then down-regulating the expression of RAGE in tissue of pancreas and kidney, ultimately mitigating the injury of pancreas and kidney. |
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