The Pharmacodynamics And Mechanism Of Hugan Buzure Granule On Anti-Acute Kidney Injury In Mice

OBJECTIVE: To study the pharmacodynamics and mechanism of Hugan Buzure Granule on anti-acute kidney injury in mice,and to provide scientific basis for its development and application in the treatment of nephropathy.METHODS: Male Balb/c mice were intraperitoneally injected with cisplatin(13 mg/kg)to establish the model of acute kidney injury(AKI)induced by cisplatin.The low(1.6 g/kg),medium(3.2 g/kg)and high(6.4 g/kg)dose groups of Hugan Buzure Granules(HBG)were given by gavage,and the Shenyan Kangfu Pian(SKP,1.2 g/kg)was used as the positive control.To observe the effect of HBG on the growth state and behavior of mice;to weigh the kidney and liver of mice,calculate the organ coefficient and observe the appearance of the kidney;at the same time,to observe the pathological changes of the kidney tissue of mice in each group by H&E staining;to determine the content of serum creatinine and urea nitrogen by biochemical kit.To further study the mechanism of HBG on AKI in mice,the amount of ROS in kidney tissue was observed by using ROS frozen section;the apoptosis of cells in kidney tissue was observed by using apoptosis staining section;MDA,GSH,CAT and SOD related indicators of oxidative stress were detected by biochemical kit;the levels of serum inflammatory factors IL-6,TNF-α and IL-1β were detected by ELISA kit;immunohistochemistry and WB were detected The protein expression of NLRP3/Caspase-1 signaling pathway and TLR4/NF-κB signaling pathway was detected.RESULTS: The content of chlorogenic acid,quercetin,kaempferol,apigenin and quercitrin in HBG was determined by HPLC.Except for the Control group,the other groups of mice were in low spirits,dark hair and other adverse conditions,HBG groups improved mice in low spirits,dark hair and other adverse conditions.Compared with the Control group,the kidney coefficient of Cisplatin group was significantly increased(P<0.01),and HBG group significantly decreased the kidney coefficient of mice(P<0.01).Compared with the Control group,the normal structure of renal tissue was obviously destroyed,showing many large vacuoles,and the arrangement of renal tubular cells was scattered,the histological characteristics of renal injury in HBG were significantly improved,the vacuoles decreased,and the arrangement of renal tubular cells tended to be neat.Compared with the Control group,the levels of Cr and BUN in serum of mice in Cisplatin group were significantly increased(P<0.01),and the levels of Cr and BUN in serum of mice in HBG groups and SKP groups were significantly decreased(P<0.01);The results of ROS staining showed that the fluorescence intensity of renal sections in Cisplatin group was significantly higher than that in Control group,and the fluorescence intensity of renal sections in HBG groups was significantly lower than that in Cisplatin group.Compared with Control group,the content of MDA in renal tissue of mice in Cisplatin group was significantly increased(P<0.01),the content of GSH was significantly decreased(P<0.05),the content of MDA in renal tissue of mice in HBG groups was significantly decreased(P<0.01),while the content of GSH was increased.The activities of CAT and SOD in Cisplatin group were significantly decreased(P<0.01),and the activities of CAT and SOD in HBG groups were significantly increased(P<0.01).Apoptosis staining showed that the positive expression of apoptosis in Cisplatin group was significantly higher than that in Control group,and the positive expression of apoptosis in HBG groups was significantly lower than that in Cisplatin group.Compared with the Control group,the serum levels of TNF-α,IL-1β and IL-6 in Cisplatin group were significantly increased(P<0.01),and the serum levels of TNF-α,IL-1β and IL-6 in HBG groups were significantly decreased(P<0.01 or P<0.05).Compared with the Cisplatin group,the expressions of NLRP3,Caspase-1,IL-18,IL-1β,TLR4,NF-κB,IL-6 and TNF-α were down regulated in HBG groups.CONCLUSION: HBG has a certain protective effect on AKI mice;HBG can protect AKI mice by reducing oxidant stress;HBG can reduce the AKI in mice by inhibiting the apoptosis of tissue cells;HBG can reduce the content of inflammatory factors in serum to alleviate AKI in mice;HBG can inhibit NLRP3/Caspase-1 signaling pathway and TLR4/NF-κB signaling pathway to prevent AKI in mice.This study clarified the effect of traditional Uygur medicine HBG on AKI,preliminarily clarified the mechanism of HBG on AKI in molecular mechanism,and provided scientific basis for its clinical application.