| Objective:Acute kidney injury(AKI)refers to a group of clinical syndromes with sudden decline in kidney function caused by one or more reasons,with or without oliguria or anuria.It has the characteristics of high prevalence and high mortality;At the same time,It is also an important cause of chronic kidney disease and chronic renal failure,but there are currently no specific therapeutic drugs,especially molecular therapeutic targets with targeted therapy functions.In this study,a model of acute kidney injury was established by injecting cisplatin injection into the abdominal cavity of mice,giving Astragalus propinquus Schischkin and Panax notoginseng compound(A&P)intragastric treatment,and then observing the pathological changes of the mice’s kidneys and the changes in the secretion and expression of inflammatory factors.In studying the regulation effect of Astragalus propinquus Schischkin and Panax notoginseng compound on renal inflammation in AKI mice and possible targeted molecular mechanisms,it provides new reference ideas and targets for the treatment of AKI.Methods:In our study,the experiment was designed as in vivo experiment and in vitro experiment.In vivo experiment: eighteen male rats were randomly divided into three groups: control group(n=6),cisplatin induced acute kidney injury model group(n=6),cisplatin induced acute kidney injury + A&P treatment group(n=6).One hour before the intraperitoneal injection of cisplatin(20 mg/kg),86.746 mg of A&P was given by gavage.Once a day,for three consecutive days.After three days,the mice were killed to make samples.In vitro experiment: low expression,over expression and blank expression models of LncRNA9884 were established by LncRNA9884 kit in vitro.After the tubular epithelial cells resuscitated,the seeding plate was formed,On the second day,LncRNA9884 was transfected,and on the third day,A&P(250 μg/m L),one hour later,adding LPS(2 μ g/ ml),to induction modeling.Control group,LPS group,LPS + A&Pgroup,LncRNA9884 overexpression group,LPS+ LncRNA9884 overexpression group,LPS + LncRNA9884 overexpression +A&P group,LPS + LncRNA9884 low expression group,LPS + LncRNA9884 low expression + A&P group,si NC group,LPS + si NC group.In this study,creatinine and urea nitrogen kits were used to detect the changes of renal function,he staining,PAS determination of chromosome and renal tubular injury score were used to determine the pathological changes of kidney,immunohistochemistry,RT-PCR and ELISA were used to detect the secretion and expression of inflammatory factors in renal tissue and renal tubule epithelial cells,and Western blot was used to detect p-YAP and p-NF-κB(P65)、IL-1、IL-6、TNF-αThe changes of relative contents of the proteins were also observed.Results:1.In vivo experiments,we found that compared with the control group,the renal tubular injury score of the model group was significantly increased(P<0.0001),the glycogen complex deposition was also significantly increased,and the renal function indexes,serum creatinine and urea nitrogen were significantly increased(P<0.01);After treatment with A&P for 3 days,renal tubular injury score was significantly reduced(P<0.01),glycogen load deposition was significantly reduced,and renal function was significantly improved(P<0.05).2.In vivo test,Rt-PCR and ELISA results showed that compared with the control group,LncRNA9884 and IL-1 in AKI model group were significantly increased(IL-1β、IL-6、TNF-α)After treatment with A&P for3 days,the inflammatory factors decreased significantly(P< 0.05).3.The results of Western blotting in vivo showed that,compared with the normal group,the expression of(IL-1β、IL-6、TNF-α)in the model group was significantly higher;also p-YAP,NF-κB(p65)and Kim-1 protein increased significantly in kidney tissue(P<0.01),and decreased significantly after treatment with A&P(P<0.05).In vitro,A&P significantly improved the inflammatory morphology of renal tubular epithelial cells induced by LPS.Compared with the control group,lncrna-9884 and Inflammatory factors(IL-1β、IL-6、TNF-α)in the over expre-ssion group were significantly increased in renal tubular epithelial cells(P<0.05).Inflammatory factors(IL-1β、IL-6、TNF-α)in inflammatory group,The expression was significantly higher than that of the control group(P<0.05);Expression of Inflammatory factors(IL-1β、IL-6、TNF-α)in LPS +LncRNA9884 over expression group,The expression was significantly higher than that in inflammatory group(P< 0.05);After treatment with A&P,the inflammation of LPS + A&P group and LPS + LncRNA9884 overexpression group + A&P group was significantly improved(P < 0.05),but the treatment effect of the latter was worse than that of the former(P<0.05).In vitro PCR results showed that A&P regulated the expression of LncRNA9884 and affected the inflammation of renal tubular epithelial cells.Expression of Inflammatory factors(IL-1β、IL-6、TNF-α)in LPS + LncRNA9884 low expression group was lower than that of the control group,the difference was statistically significant(P < 0.05);After treatment with A&P,the curative effect was better than that of LPS + A&P group(P < 0.05);In si NC group,the level of IL-1β、IL-6、TNF-α,the expression and secretion had no statistical significance compared with the control group;in LPS + si NC group,expression of Inflammatory factors(IL-1β、IL-6、TNF-α)was no significant difference with the Inflammation group.Conclusion:1.A&P can improve the renal function of AKI mice induced by cisplatin,reduce the pathological changes,and effectively inhibit the inflammation of renal tissue.Therefore,A&P can improve the kidney injury and inflammation of AKI mice.2.The mechanism of A&P in improving acute kidney injury in mice is to regulate LncRNA9884/YAP/NF-κB signal axis and reduce the secretion and expression of inflammatory factors downstream. |
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