| Objective: To observe the therapeutic effect of Jianzhong Bushen Xiaozhengtang on unilateral ureteral obstruction(UUO)mice based on Jianzhong Bushen Xiaozhengtang,as well as the regulatory effect on the apoptosis pathway of mitochondria,and further explore the mechanism of Jianzhong Bushen Xiaozhengtang in improving RIF,which provides experimental basis for clinical application of this prescription,and also offers fresh basis and pointss for the prevention and therapia of CKD by traditional Chinese medicine.Methods: The UUO mouse model was used,with different doses of Jianzhong Bushen Xiaozheng Decoction intervention and Niaoduqing granules as positive drug control.The mice in each group were gavaged from the 3rd day after the model was made,and 5/6 mice were randomly selected from the 14 th,21st and 28 th days after gavage for sampling.Observe the changes of general conditions of mice in different periods,use automatic biochemical analyzer to detect serum Scr and BUN,HE staining to survey the pathologic varieties of nephridial histo,and TUNEL staining to survey the apoptosis of nephridial tubular epithelial cells,Western blot and RT-PCR were used to check the expression of Bcl-2,Bax and Apaf-1 in the kidney tissue of the obstructive side.Results:(1)General situation: The body weight of the UUO model group mice significantly decreased compared to the sham surgery group(P<0.01);Compared with the UUO model group,each treatment group was able to improve the general condition of mice and gradually increase their body mass.On the 28 th day after modeling,the body mass of mice in the high,medium,and low dose groups of traditional Chinese medicine and the Niaoduqing group significantly increased compared to the UUO model group(P<0.01);(2)Scr and BUN detection: The concentration of Scr and BUN in the UUO model group mice significantly increased compared to the sham operation group at the same period(P<0.05,P<0.01);Compared with the UUO model group at the same period,the decrease in Scr concentration in each treatment group of mice was not significant(P>0.05)two weeks before administration.Starting from the 21 st day,the Scr concentration in each treatment group of mice was significantly reduced(P<0.05);Compared with the UUO model group at the same period,the BUN concentration of mice in the high and medium dose groups of traditional Chinese medicine and the Niaoduqing group significantly decreased 14 days after modeling(P<0.01,P<0.01,P<0.05);Starting from the 21 st day of administration,the BUN concentration in all treatment groups of mice significantly decreased,with statistical significance(P<0.05,P<0.01);(3)HE staining: The degree of renal tissue damage in mice treated with Jianzhong Bushen Xiaozheng Tang was reduced to varying degrees compared to the UUO model group;(4)TUNEL staining:Compared with the sham surgery group,the positive staining of RTECs in the UUO model group was significantly increased(P<0.01);After administration,the apoptosis of RTECs in each treatment group improved to varying degrees,with a decrease in positive staining.Compared with the UUO model group at the same period,the apoptosis of RTECs in the high-dose and Niaoduqing groups of traditional Chinese medicine was significantly reduced(P<0.01);The number of apoptosis in RTECs in the low-dose group of traditional Chinese medicine at 14 days after modeling showed no significant change compared to the same period of UUO model group.After 21 days of modeling,the number of apoptosis in RTECs was significantly lower than that in the same period of UUO model group(P<0.05,P<0.01);(5)Western Blot: Compared with the sham surgery group,the expression of Bcl-2in the UUO model group was significantly reduced(P<0.01),while the expression of Bax and Apaf-1 was significantly increased(P<0.01);Compared with the UUO model group,the high-dose group of traditional Chinese medicine and the Niaoduqing group showed a significant increase in Bcl-2 expression(P<0.01),while the expression of Bax and Apaf-1decreased significantly(P<0.01);On the 14 th day after modeling,there was no significant difference in the expression of Bcl-2,Bax,and Apaf-1 between the medium and low dose groups of traditional Chinese medicine and the UUO model group(P>0.05).However,with the prolongation of gavage time,the expression of Bcl-2 in the medium dose group of traditional Chinese medicine significantly increased compared to the UUO model group(P<0.01),while the expression of Bax and Apaf-1 significantly decreased(P<0.01);Among the treatment groups of traditional Chinese medicine,the high-dose group of traditional Chinese medicine showed higher expression of Bcl-2 protein in the kidney tissue of mice(P<0.01);(6)q RT PCR: The expression of Bcl-2 m RNA in the kidney tissue of UUO model group mice was significantly reduced compared to the sham operation group at the same period(P<0.01),while the expression of Bax and Apaf-1 m RNA was significantly increased(P<0.01);The expression levels of Bcl-2 m RNA in the high-dose group of traditional Chinese medicine and the Niaoduqing group were significantly increased compared to the UUO model group at the same period(P<0.01),while the expression levels of Bax and Apaf-1 m RNA were significantly reduced(P<0.01);There was no significant difference in the expression of Bcl-2 in the middle dose group of traditional Chinese medicine in the early stage(P>0.05),but as the intervention time prolonged,the expression of Bcl-2 m RNA in the kidney tissue of mice in the middle dose group of traditional Chinese medicine increased significantly(P<0.05,P<0.01)at 21 and28 days after modeling;The Bax m RNA level of mice in the medium dose group of traditional Chinese medicine was significantly reduced compared to the UUO model group at the same period(P<0.01);Although the expression of Apaf-1 m RNA in the kidney tissue of mice in the medium dose group of traditional Chinese medicine was lower than that of the UUO model group at the same time,there was no significant difference and no statistical significance(P>0.05);Compared with the high-dose group of traditional Chinese medicine during the same period,the expression of Bcl-2 m RNA in the low-dose group of traditional Chinese medicine was significantly lower than that in the high-dose group(P<0.01).The middle-dose group of traditional Chinese medicine only showed a significant difference from the high-dose group at 14 days(P<0.01).The expression of Bax m RNA in the low-dose group of traditional Chinese medicine was significantly higher than that in the high-dose group(P<0.01),and there was no significant difference between the other treatment groups and the high-dose group of traditional Chinese medicine(P>0.05)The low-dose group was significantly higher than the high-dose group(P<0.01,P<0.05),while the Niaoduqing group was higher than the high-dose group,but there was no significant difference and no statistical significance(P>0.05).Conclusion: Cell apoptosis mediated by mitochondrial apoptosis pathway plays an important role in the process of renal interstitial fibrosis in UUO mice.Jianzhong Bushen Xiaozheng Decoction may inhibit the apoptosis of renal tubular epithelial cells by down-regulating the expression of Apaf-1 and Bax and up-regulating the expression of Bcl-2,delay the progression of RIF,and improve renal function. |
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