| Objective:To study the process that Jianzhong Bushen Xiaozheng Decoctionregulatesthedynamicbalanceofmatrix metalloproteinase-9/matrix metalloproteinase inhibitor-1（MMP-9/TIMP-1）to reduce the effect of extracellular matrix（ECM）against renal interstitial fibrosis（RIF）in UUO model mice,clarify the mechanism of action of this prescription and Jianzhong Bushen method,further determine the effectiveness and safety of Jianzhong Bushen Xiaozheng Decoction on RIF,and provide scientific basis and treatment ideas for clinical use.Methods:120 mice were randomly divided into sham operation group（sham group）,unilateral ureteral obstruction（UUO）model group,Niaoduqing group,Jianzhong Bushen Xiaozheng Decoction high,middle and low dose groups,and UUO animal model was used.The drug was administered by gavage 3 days after the modeling operation.The UUO model group and the sham group were given the same amount of distilled water by gavage every day.The low,medium and high dose groups of Jianzhong Bushen Xiaozheng Decoction were given the water solution of Jianzhong Bushen Xiaozheng Decoction by gavage at the rate of6,12,24g·kg-1·d-1,respectively,and the urea clear group was given the water solution of Niaoduqing granules by gavage at the rate of 6.2 g·kg-1·d-1.The morphological changes of mice after 14,21,and 28 days were observed,and the general physical signs of mice and various indicators after the treatment of obstruction were observed.After the administration,samples were taken and the pathological changes of kidney tissue were observed by hematoxylin-eosin staining（HE staining）;The contents of serum creatinine（Scr）and Blood urea nitrogen（BUN）in mice were detected by automatic biochemical instrument;Enzyme-linked immunosorbent assay（ELISA）was used to detect the expression of MMP-9,TIMP-1 and type IV collagen（Col-Ⅳ）in mouse serum.Results:1.General physical signs of miceAfter modeling,the mice in each medication group and UUO group have dark skin color,lazy posture,insensitive response,and even limb edema of different degrees,which are more consistent with the characteristics of CRF.After medication（Niaoduqing,Jianzhong Bushen Xiaozheng Decoction,each dose group）,the above symptoms were improved to varying degrees,such as the hair color was gradually clean and shiny,the body mass was slightly increased,the body posture was flexible,the reaction was rapid,the degree of tarsal swelling was reduced,and there were no other obvious adverse reactions.2.Pathological staining results of mouse kidney tissueIn sham group,the epithelial cells of renal tubules were closely arranged,the cell size was normal,the nucleolus was clear,the structure of renal tubules was complete,and the number was abundant.The renal tissue structure of UUO group was abnormal,and with the increase of obstruction time,renal tubule expansion,inflammatory cell infiltration,vacuolar degeneration of renal tubule epithelial cells and RIF appeared.The degree of renal tissue injury in each treatment group was less than that in the UUO group,and after the intervention of Jianzhong Bushen Xiaozheng Decoction,the degree of cell edema and RIF in each dose group of traditional Chinese medicine decreased in a dose-dependent manner.3.Test results of biochemical indicators of mouse kidney functionCompared with Sham group,Scr and BUN in UUO group were significantly higher on the 14th,21st and 28th day respectively（P<0.05）.After treatment,the biochemical indexes of the treatment group at the same time were lower than those of the UUO group in varying degrees,and the treatment effect of Chinese medicine in the high-dose group was obvious,with the most significant decrease（P<0.05）.At the 14th day,there was no significant difference in the content of Scr between the low-dose group of traditional Chinese medicine and the UUO group（P>0.05）.4.Detection results of MMP-9,TIMP-1 and Col-Ⅳin mouse serumCompared with the expression of Sham group,the content of MMP-9 protein in UUO group decreased significantly（P<0.05）;After treatment,there were different degrees of elevation in each treatment group on the 21st and 28th days（P<0.05）,and the expression was most obvious in the high-dose group of traditional Chinese medicine;On the 14th day,there was no significant increase in Niaoduqing group and low-dose Chinese medicine group（P>0.05）.The content of TIMP-1 in UUO group was significantly higher than that in sham group at the same period（P<0.05）.After treatment,the content of TIMP-1 in each treatment group decreased significantly at the same period（P<0.05）.Compared with UUO group,with the increase of MMP-9 content and the decrease of TIMP-1 level,the ratio of MMP-9/TIMP-1 in each treatment group gradually increased,and the effect of high-dose Chinese medicine group was the best（P<0.05）.On the 14th day,the ratio of Niaoduqing group and low-dose Chinese medicine group was not significantly higher than that of UUO group（P>0.05）.The results of Col-Ⅳin UUO group were significantly higher than those in sham group in three periods（P<0.05）;The protein expression in each treatment group decreased on the 21st and 28th days（P<0.05）,and there was no significant difference between Col-Ⅳin the Niaoduqing group and the UUO group on the 14th day（P>0.05）.Conclusion:Jianzhong Bushen Xiaozheng Decoction has a good effect on improving the RIF and renal tissue of UUO model mice.Its mechanism may be related to promoting the degradation of Col-Ⅳand reducing the synthesis of ECM deposition by up-regulating MMP-9,down-regulating the expression of TIMP-1 and restoring the steady-state balance between MMP-9 and TIMP-1through Jianzhong Bushen Xiaozheng Decoction.This study provides an experimental basis for clinical treatment of CRF with Jianzhong Bushen Xiaozheng Decoction. |
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