| Objective: Through experimental studies,observe the effects of Shenfu Decoction on renal function,kidney tissue morphology and TGF-β1/Smads signaling pathway in CKD rats,in order to explore the mechanism of Shenfu Decoction in delaying CKD progression and anti-renal fibrosis.Methods: In this study,60 SPF SD rats were selected,and after 7 days of adaptive feeding,they were randomly divided into 12 blank groups and 48 model groups.The model was constructed by adenine intragastric administration to establish a CKD rat model.After the model was completed,3 rats were randomly killed in the blank group and the model group.HE staining was used to verify the success of the model.After verification,the building blocks were randomly divided into model group,Niaoduqing granule group,Shenfutang low-dose group,Shenfutang medium-dose group,and Shenfutang high-dose group,9 in each group.Rats in each group were intervened for 28 days.During this period,attention was paid to observe changes in the general state and weight of rats;after 28 days,blood and kidney samples were collected for detection;HE and Masson staining methods were used to observe changes in kidney morphology;ELISA was used to detect CKD Expression of TGF-β1,CTGF,TNF-α,Smad7 in rat serum;Western Blot method was used to detect the expression level of TGF-β1,p-Smad2/3,Smad7 protein in rat kidney tissue;immunohistochemical staining To detect the expression levels of TGF-β1,Smad2,CTGF,IL-8,TNF-α and Smad7 in kidneys of CKD rats;use RT-PCR method to detect the relative expression of TGF-β1m RNA,Smad2 m RNA and Smad7 m RNA in CKD rat kidneys The amount of expression.Result:1.The general state of rats in each dose group of Shenfu Decoction has improved to varying degrees,and it is significantly better than the model group in terms of body weight,urine output,diet and mental state.The general status of Shenfutang high-dose group and Niaoduqing granule groupwas slightly better than that of Shenfutang medium-and low-dose group.2.Biochemical and blood routine results showed that: Compared with the blank group,the serum creatinine and urea nitrogen content of the model group were significantly increased(P<0.01);compared with the model group,the serum creatinine,The urea nitrogen content decreased in varying degrees(P<0.05).The levels of blood creatinine and urea nitrogen in Shenfutang high-dose group in each treatment group were similar to those in Niaoduqing granule group,and were superior to Shenfutang medium-and low-dose group.Compared with the blank group,the hemoglobin content of the model group decreased significantly(P<0.01);compared with the model group,the hemoglobin of each treatment group of Shenfu Decoction increased to varying degrees.The hemoglobin recovery level of the high-dose Shenfu Decoction group was similar to that of the Niaoduqing Granule group in each treatment group,and it was better than that of the medium-and low-dose Shenfu Decoction group.3.Pathological results showed that the degree of fibrosis in kidney tissue of CKD rats in each dose group of Shenfu Decoction improved to varying degrees.Compared with the model group,the percentage of vascularized area in each dose group of Shenfu Decoction decreased to varying degrees,and the difference has statistics Academic significance(P<0.05).4.ELISA test results showed that: Compared with the blank group,the expression of TGF-β1,CTGF,and TNF-α in the serum of the model group were significantly increased(P<0.01);compared with the model group,each dose group of Shenfu Tang It can reduce the expression of TGF-β1,CTGF and TNF-α in rat serum to a certain extent(P<0.05).5.Western Blot test results showed that: compared with the blank group,the expression of TGF-β1 and p-Smad2/3 in the kidney tissue of the model group were significantly increased(P<0.01);compared with the model group,Shenfu Tang The expression levels of TGF-β1 and p-Smad2/3 in the kidney tissues of rats in the dose group all decreased to varying degrees(P<0.05).Compared with the blank group,the expression of Smad7 in the kidney tissue of the model group was significantly decreased(P<0.01);compared with the model group,each dose group of Shenfu Decoction could increase the expression of Smad7 in the kidney tissue of the rat to a certain extent(P<0.05).6.The results of immunohistochemistry showed that: compared with the blank group,the expression of TGF-β1,Smad2,CTGF,IL8,and TNF-α in the kidney tissues of the model group were significantly increased(P<0.01);compared with the model group In contrast,the expressions of various inflammation indexes in each treatment group of Shenfu Decoction decreased to varying degrees(P<0.05).Compared with the blank group,the expression of Smad7 in the kidney tissue of the model group was significantly decreased(P<0.01);compared with the model group,the expression of Smad7 in each treatment group of Shenfu Decoction increased to varying degrees(P<0.05).7.RT-PCR test results showed that: Compared with the blank group,the relative expression levels of TGF-β1m RNA and Smad2 m RNA in the kidney tissues of the model group were significantly increased(P<0.01);compared with the model group,each treatment group of Shenfu Decoction The relative expression levels of TGF-β1m RNA and Smad2 m RNA were down-regulated to varying degrees(P<0.05).Compared with the blank group,the relative expression of Smad7 m RNA in the kidney tissue of the model group was significantly down-regulated(P<0.01);compared with the model group,the expression of Smad7 m RNA in each treatment group of Shenfutang was up-regulated to varying degrees(P<0.05).Conclusion:1.Shenfu Decoction can improve the general state of CKD rats,increase hemoglobin,improve renal function and kidney pathological damage.2.The TGF-β1/Smads signal transduction pathway may be one of the mechanisms of Shenfu Decoction against renal interstitial fibrosis.Based on this transduction pathway,Shenfu Decoction can effectively reduce theexpression of TGF-β1,CTGF,TNF-α and other pro-fibrotic factors in the serum and kidney of CKD rats,increase the expression of anti-fibrosis factor Smad7,and then reduce the synthesis of extracellular matrix And apoptosis.3.Shenfu Decoction has a certain anti-renal interstitial fibrosis effect in delaying the process of CKD.The high-dose group of Shenfu Decoction has better effect,suggesting that the amount of Chinese medicine is positively correlated with the efficacy in this study. |
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