| Objective:Renal fibrosis is a dynamic process that originates from various pathogenic factors and involves various aspects such as inflammatory response and oxidative stress.It eventually will develop into chronic renal failure.In this paper,we studied the molecular mechanism of kangxianling decoction resisting renal fibrosis from RAS system and TGF-?1/Smad signaling pathway,and provided theoretical basis for multi-target and multi-directional treatment of Traditional Chinese Medicine.Methods:(1).According to the Platt method,renal fibrosis model was induced through5/6 nephrectomized rats.Two weeks after surgery,blood was collected from the eyes and blood creatinine was measured.After successful modeling,according to the serum creatinine level,5/6 nephrectomized renal fibrosis group was divided into model group,kangxianling decoction group and losartan potassium group.So there was no statistical difference between the different groups(P>0.05).Kangxianling decoction group was kangxianling decoction,the concentration was 21g·kg-1,and losartan potassium group was given losartan potassium,the drug concentration was 33.3 g·kg-1,once a day,every time 10 ml·kg-1 was administered.The control group,the sham operation group,and the model group were all fed freely and given saline 10 ml·kg-1 once a day.Rats were continuously intragastrically administrated for 24 weeks,Scr,Bun,and 24h Upro were measured at 8th,16th,and 24th weekends.At the same time,the pathological changes were observed by HE and Masson staining.The expressions of CTGF,Col-I,Col-III,ACE1 and AT1R in renal tissues were detected by immunohistochemistry.The expressions of ACE1,Ang ?,AT1R,TGF-?1,Smad2/3 and Smad7 were detected by Western Blot.The expressions of ACE1,Ang ?,AT1R,TGF-?1,Smad2/3 and Smad7were detected by real-time fluorescent quantitative PCR.(2).Rat mesangial cells(HBZY-1)were randomly divided into control group,model group(Ang II stimulation group),low concentration kangxianling decoction group,high concentration kangxianling decoction group,and losartan potassium group.The kangxianling decoction group and the losartan potassium group were pretreated with 100?g/ml kangxianling decoction,200?g/ml kangxianling decoction and 10?M losartan potassium for 0.5 h.Then,the model group,100?g/ml kanxianling decoction group,200?g/ml kangxianling decoction group,and losartan potassium group were all stimulated with 100 nm Ang II for 24 hours,and finally the cells were harvested.The concentration of Ang II cytokines in each group was detected by Elisa,and the expression of TGF-?1,Smad2/3 and Smad7 in the mesangial cells induced by Ang II stimulation was detected by Western Blot.Result:(1).Kangxianling can significantly improve the levels of Scr,Bun and 24h Upro in rats with chronic renal failure,and effectively improve renal function and delay the progression of chronic renal failure.(2).Kangxianling decoction can significantly reduce the protein expressions of ACE1,Ang II,AT1R,Tgf-?1 and Smad2/3 in renal tissue of rats with chronic renal failure,increase the protein expression of Smad7,thereby inhibiting renal fibrosis.(3).Kangxianling decoction can reduce the expressions of ACE1 m RNA,Ang II m RNA,AT1 m RNA,Tgf-?1 m RNA,Smad2 m RNA and Smad3 m RNA in kidney tissue,and increase the expression of Smad7 m RNA.(4).Kangxianling decoction can inhibit the proliferation of HBZY-1 cells induced by Ang II,inhibit the transduction of Tgf-?1/Smad signaling pathway,decrease the expression of Tgf-?1 and Smad2/3 protein in Tgf-?1/Smad signaling pathway,and increase the Smad7 protein.There was a dose-dependent relationship between the concentration of kangxianling decoction and protein expression.The higher the concentration of kangxianling decoction,the stronger the inhibition and the lighter the degree of renal fibrosis.(5).Kangxianling decoction has the function of protecting kidney function and inhibiting renal fibrosis.The main mechanism may be related to inhibition of ACE-Ang II-AT1R signaling pathway activation and Tgf-?1/Smad signal transduction. |
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