| Objectives:The study aimed to explore whether the of alcohol-induced rat kidney damage is related to mitochondrial oxidation and TGF-?/Smads signaling pathways,and to studies of the effect and mechanism of traditional Chinese medicine compound Kangshen Prescription?KSP?on alcoholic kidney damage.Methods:A wistar rat model?35 rats?was given alcohol(400g·L-1,20ml·kg-1)that is diluted from 56°alcohol?441.84g/L?two times a day in the course of 6 weeks,the blank group was given 0.9%sodium chloride(40ml·kg-1·d-1)by gavage?15 rats?.After exposed to alcohol for6 weeks,blood urea nitrogen,serum creatinine,and kidney index were measured in control and blank groups?each of 5 rats?,and morphology of the kidneys were observed.After the model was established,the control and blank groups continues to be given 0.9%sodium chloride(20ml·kg-1·d-1,each of 10 rats)for gavage,KSP and Enalapril?Positive control?group respectively were given kangShen Prescription(12ml·kg-1·d-1,n=10)and Enalapril solution(10ml·kg-1·d-1,n=10)by gavage for 6weeks.After 2 weeks and 6weeks of treatment,blood of the rats was collected,and the kidneys were stored.Kidney index was calculated,renal function was tested by BUN and Scr separately,pathological changes of renal tissue were also observed.The apoptosis-related proteins?Cyt-c,Caspase-8,Caspase-9,Bax,Bcl2 and p53?in the kidney were analysed by WB,the protein expression of CD34 in endothelial cell was determined by immunofluorescence,and the gene expression levels of TGF-?,Smad3,Smad7 and Col1a1 in renal interstitial fibrosis were detected by RT-PCR.Results:1)After exposed to alcohol for 6 weeks,compared with blank group,The renaltubules and glomeruli of the control group showed obvious Collagen with a large number of inflammatory cell infiltration,and it can be obviously seen that a small number of sprouting blood vessels extending toward the medulla around the capillaries in the renal cortex.The RI and TIS have statistical differences?P<0.05?.2)After 2 weeks of treatment,compared with the control group,the expression of TGF-?and Smad3 genes were down-regulated,the Gene of Smad7 was up-regulated?P<0.05?in KSP group,and the expression level of TGF-?,Smad3,and Col1a1genes in enalapril group were down-regulated?P<0.05?.3)After 6 weeks of treatment,compared with blank group,the level of CD34,Cyt-c,Caspase-8,Caspase-9,Bax,Bcl2,and p53 proteins in renal tissue were elevated of the control group?P<0.05?.meanwhile,the gene expression of TGF-?,Smad3,and Col1a1 were up-regulated?P<0.01?,and the Smad7 gene expression was down-regulated?P<0.01?.After treatment,the expression of Collagen around the renal tubules were all decreased,the renal index,Scr,and BUN decreased significantly in KSP and enalapril groups.the same that the level of proteins?CD34,Cyt-c,Caspase-8,Caspase-9,Bax,and p53?in the kidney tissue are significantly reduced?P<0.05?,while the level of Bcl-2 rised?P<0.01?.The level of genes?TGF-?,Smad3,and Col1a1?in KSP and enalapril groups were down-regulated,but the Smad7 gene showed up-regulated?P<0.01?.Conclusions:1.Alcohol-induced rat kidney damage may be related to mitochondrialoxidation-activated JNK pathway.2.Alcohol-induced renal vascular endothelial cell proliferation and renal interstitial fibrosis may be related to TGF-?/Smad signaling pathways.3.KSP may relieve the development of alcoholic kidney damage by regulating TGF-?/Smad signaling pathway and inhibiting oxidation reaction,apoptosis and inflammatory response. |
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