| Background:Medicine is one of the main weapons for human beings to overcome diseases.Drug progress is one of the necessary conditions for human survival and development.However,compared with the therapeutic effect,the toxic and side effects of drugs are often ignored.Drug-induced renal injury is one of the most common and representative diseases in drug-induced diseases.Clinical data show that about twenty-five percent of patients with renal failure are related to drug-induced renal injury.In the future,drug-induced renal injury will become an increasingly serious economic and major public health problem.Every rigorous medical worker and manager,from theory to practice,should pay the same attention to the safety and effectiveness of clinical medication.Most drugs may have different degrees of adverse reactions.However,generally,mild to moderate adverse reactions have mild symptoms and short duration,and fail to form the corresponding clinical process.This situation can not be called "drug-induced disease".Only those with significant clinical symptoms or obvious abnormal laboratory test results and corresponding clinical process are called drug-induced diseases.Drug-induced renal injury or toxic nephropathy is not an independent disease,but the general name of a large class of diseases.A kind of disease with corresponding clinical manifestations caused by drug-induced renal structure or function damage is called drug-induced renal injury,which is mainly manifested in nephrotoxicity and allergy.The anatomical and physiological characteristics of the kidney make the kidney very sensitive to the toxicity of drugs and easy to cause damage.The weight of the kidney is less than one percent of the body weight,while 20%?30% of the blood from the heart flows into the kidney,causing the kidney to be exposed to a large amount of drugs.Secondly,the countercurrent doubling mechanism of renal medulla and the activity of drug metabolizing enzymes in proximal convoluted tubules are easy to be inhibited by toxic drugs.In addition to the above important reasons,the massive ATP consumption of kidney and the hemodynamic characteristics of kidney have become the basis of renal injury.Drug-induced renal injury is one of the most common and representative diseases in drug-induced diseases.Clinical data show that about twenty-five percent of patients with renal failure are related to drug-induced renal injury.In the future,drug-induced renal injury will become an increasingly serious economic and major public health problem.Drug-induced renal injury can be classified into "urine turbidity","edema","Guan Ge","closure of scrofula","low back pain" and "hematuria" from the perspective of TCM.The main cause of this disease is that the drug poisons hurt the kidney,so its pathological nature belongs to evil excess and positive injury.It is empirical in the initial stage,and then mostly deficiency.Blood stasis,damp heat and toxin evil run through drug-induced renal injury.Actively using the methods of promoting blood circulation and removing blood stasis,promoting dampness and resolving phlegm and detoxification is of great positive significance for the prevention and treatment of drug-induced renal injury.According to the theories of TCM,we believe that Reynoutria japonic,which is widely used in the treatment of acute and chronic renal injury in clinic,may play an important role in the prevention and treatment of drug-induced renal injury.We believes that the generation of reactive ROS is the core link in the pathogenesis of drug-induced renal injury,and the production of ROS is closely related to the aging related gene SIRT1.Reynoutria japonic has definite curative effect on drug-induced renal injury.Resveratrol is the main extract of Reynoutria japonic.SIRT1 has strong antioxidant damage activity,can change the level of SOD and MDA.Resveratrol is a natural SIRT1 activator.Therefore,we propose a scientific hypothesis that SIRT1 may be involved in the occurrence of drug-induced renal injury,SIRT1/SOD/MDA may be an effective target for the prevention and treatment of drug-induced renal injury,and Reynoutria japonic and resveratrol may play a role in the treatment of drug-induced renal injury through oxidative stress.Basic research Objective:RT-PCR,UV spectrophotometry,immunohistochemistry and PAS staining of pathological sections were used to understand the effect of drug intervention on SIRT1/SOD/MDA in rat kidney induced by doxorubicin,and to explore the mechanism of the tested drug from the aspect of oxidative stress.Methods:After three days of adaptive feeding,fifty male SD rats were randomly divided into five groups: blank group,model group,Reynoutria japonic group,resveratrol group and benazepril hydrochloride group,with ten rats in each group.The model group,Reynoutria japonic group,resveratrol group and benazepril hydrochloride group used a one-time tail vein injection of 7.5mg/kg doxorubicin to make the rat model of drug-induced renal injury.After three days,the proteinuria test strip showed green,which was positive for proteinuria.For the rats that failed to make the model,the above methods can be repeated.In this experiment,the rats were successfully made.The blank group and model group were given distilled water 2ml/d by gavage,Reynoutria japonic group,benazepril hydrochloride group and resveratrol group were given Reynoutria japonic 10g/kg,benazepril hydrochloride0.90mg/kg/d and resveratrol 40mg/kg/d by gavage respectively,six times a week for four weeks.Rats were weighed in the early morning of the first day of each week to adjust the dose.On the morning of the seventh day of each week,urine was collected randomly with metabolic cage to measure urinary protein excretion.At the end of the 4th week,blood was collected from the hearts of all rats(rat serum was collected to detect creatinine and urea nitrogen),bilateral kidneys were taken for reverse transcription polymerase chain reaction,immunohistochemistry and PAS staining,and the levels of malondialdehyde,superoxide dismutase were detected by kit.Result:1.Changes of urine volume of rats in each group: at the first week,the urine volume of blank group and benazepril hydrochloride group was significantly higher than that of model group(P<0.05),and there was no significant difference between Res and Reynoutria japonic group and model group(P>0.05).The urine volume of blank group was significantly higher than that of benazepril hydrochloride group(P<0.05),and that of Res,Reynoutria japonic group and model group was significantly lower than that of benazepril hydrochloride group(P<0.05).At the second week,there was no significant difference between each group and the model group(P>0.05).The urine volume of blank group was significantly higher than that of benazepril hydrochloride group(P<0.05).There was no significant difference in Res,Reynoutria japonic group and model group compared with benazepril hydrochloride group(P>0.05).At the third week,the urine volume of blank group,Reynoutria japonic group and benazepril hydrochloride group was significantly higher than that of model group(P<0.05),and there was no significant difference in Res compared with model group(P>0.05).The urine volume of blank group was significantly higher than that of benazepril hydrochloride group(P<0.05),and that of model group was significantly lower than that of benazepril hydrochloride group(P<0.05).There was no significant difference between Res and Reynoutria japonic group and benazepril hydrochloride group(P>0.05).At the fourth week,the urine volume of blank group,Reynoutria japonic group and benazepril hydrochloride group was significantly higher than that of model group(P<0.05),and there was no significant difference in Res compared with model group(P>0.05).The urine volume of blank group was significantly higher than that of benazepril hydrochloride group(P<0.05),and that of model group was significantly lower than that of benazepril hydrochloride group(P<0.05).There was no significant difference between Res and Reynoutria japonic group and benazepril hydrochloride group(P>0.05).2.Body weight changes of rats in each group: at the first week,the body weight of benazepril hydrochloride group and blank group was higher than that of model group,and there was significant difference between the two groups and model group(P<0.05).There was no significant difference between Reynoutria japonic group and Res group and model group(P>0.05).The model group was significantly lower than benazepril hydrochloride group(P<0.05).There was no significant difference in blank group,Reynoutria japonic group and Res group compared with benazepril hydrochloride group(P>0.05).At the second week,the body weight of benazepril hydrochloride group,Reynoutria japonic group and blank group was higher than that of model group.There were significant differences between the three groups and model group(P<0.05),and there was no significant difference between Res group and model group(P>0.05).Res group and model group were significantly lower than benazepril hydrochloride group(P<0.05).There was no significant difference between Reynoutria japonic group and blank group and benazepril hydrochloride group(P>0.05).At the third week,the body weight of benazepril hydrochloride group and blank group was higher than that of model group,and there was significant difference between the two groups and model group(P<0.05).There was no significant difference between Reynoutria japonic group and Res group and model group(P>0.05).The blank group was significantly higher than benazepril hydrochloride group(P<0.05),the model group was significantly lower than benazepril hydrochloride group(P<0.05),and there was no significant difference between Reynoutria japonic group and Res group and benazepril hydrochloride group(P>0.05).At the fourth week,the body weight of benazepril hydrochloride group,Reynoutria japonic group and blank group was higher than that of model group.There were significant differences between the three groups and model group(P<0.05),and there was no significant difference between Res group and model group(P>0.05).The blank group was significantly higher than benazepril hydrochloride group(P<0.05),and the Res group and model group were significantly lower than benazepril hydrochloride group(P<0.05).There was no significant difference between Reynoutria japonic group and benazepril hydrochloride group(P>0.05).3.Comparison between ACR groups in rats within four weeks of treatment,the urinary microalbumin/creatinine ratio of the treatment group and the model group was higher than that of the blank group.At the first week,the urinary microalbumin/creatinine ratio of the benazepril hydrochloride group was significantly lower than that of the model group(P<0.05),and the urinary microalbumin/creatinine ratio of the model group was higher than that of the treatment group(P<0.05).Compared with benazepril hydrochloride group,the urinary microalbumin/creatinine ratio of Reynoutria japonic group was slightly higher than that of benazepril hydrochloride group at the fourth week,but there was no significant difference between the two groups,while the urinary microalbumin/creatinine ratio of benazepril hydrochloride group was lower than that of Reynoutria japonic group at the first week(P<0.05).Within four weeks,the urinary microalbumin/creatinine ratio in resveratrol group was higher than that in benazepril hydrochloride group and Reynoutria japonic group.4.Comparison of serum creatinine and urea nitrogen of rats in each group.At the fourth week of treatment,the two values of serum creatinine and urea nitrogen in the treatment group and the blank group were lower than those in the model group(P<0.05).The two values of Reynoutria japonic group were significantly lower than those of benazepril hydrochloride group and resveratrol group(P<0.05).These two values in benazepril hydrochloride group were lower than those in resveratrol group(P<0.05).5.Comparison of SOD and MDA in renal tissue of rats in each group at the end of the fourth week,the levels of SOD and MDA in renal tissue of rats were compared between groups.The concentration of SOD in model group and treatment group was lower than that in blank group,and the concentration of SOD in model group was lower than that in treatment group(P<0.05).Compared with benazepril hydrochloride group,the SOD concentration in Reynoutria japonic group was significantly higher than that in benazepril hydrochloride group(P<0.05).The concentration of SOD in resveratrol group was lower than that in benazepril hydrochloride group and Reynoutria japonic group(P<0.05).In contrast to the concentration of SOD,the results of inter group comparison showed that the MDA level of model group and treatment group was higher than that of blank group,and the MDA level of model group was higher than that of treatment group(P<0.05).The level of MDA in Reynoutria japonic group was significantly lower than that in benazepril hydrochloride group(P<0.05).The level of MDA in resveratrol group was higher than that in benazepril hydrochloride group and Reynoutria japonic group(P<0.05)6.Comparison of SIRT1 gene and protein expression in renal tissue of rats in each group at the end of the 4th week,there was no significant difference between Reynoutria japonic group(P>0.05).The SIRT1 gene and protein levels of rats in model group and treatment group were lower than those in blank group,and those in model group were lower than those in treatment group(P<0.05).Compared with benazepril hydrochloride group,SIRT1 gene and protein levels in Reynoutria japonic group were significantly higher than those in benazepril hydrochloride group(P<0.05).The SIRT1 gene and protein levels in resveratrol group were lower than those in benazepril hydrochloride group and Reynoutria japonic group(P<0.05).7.Renal histology under the light microscope,it can be seen that at 4 weeks,the kidneys of the model group showed fibrosis and hyperplasia,renal interstitial congestion,rupture and bleeding of glomerular basement membrane,a large number of inflammatory cell chemotaxis in glomerulus and interstitium,tubular proteinuria,renal tubular and glomerular atrophy,etc.The kidneys of resveratrol group and benazepril hydrochloride group had mild glomerular hemorrhage and a small amount of inflammatory cell infiltration in interstitial tissue.The pathological changes of kidney in Reynoutria japonic group were mild,followed by benazepril hydrochloride group,and the pathological changes in resveratrol group were the most serious.Conclusion:The content of MDA in the model group was significantly higher than that in the normal group,which also suggested that oxidative stress played an important role in the rat model of drug-induced renal injury.The tested drugs can up regulate the expression of SIRT1,which also suggests that the tested drugs may regulate the anti lipid peroxidation of the body by mediating SIRT1 pathway,increase the synthesis of SOD in cells,reduce the synthesis of reactive oxygen species ROS in cells,then inhibit the oxidative damage of rat kidney induced by doxorubicin,and finally realize its antioxidant physiological function.The therapeutic effect of Reynoutria japonic group is significantly better than that of resveratrol group and benazepril hydrochloride group,indicating that the effect of Reynoutria japonic on kidney is multi-target and the protective effect on kidney disease is more comprehensive,including not only anti lipid peroxidation,but also other mechanisms,which need to be further studied.Clinical research Objective:This project takes the patients with drug-induced renal injury as the research object,and studies from the following aspects: first,observe whether SIRT1/SOD/MDA is involved in the pathogenesis of DKI patients,and the relationship between disease development and SIRT1/SOD/MDA,so as to understand the role of oxidative stress in drug-induced renal injury.The follow-up basic experiment was followed by the observation and study of drug intervention.Methods:Twenty drug-induced renal injury patients admitted to our Hospital from April 2020 to April 2021 were selected as the observation group.In addition,ten normal samples of the same age group and physical examination in our hospital were collected as the normal control group.The basic information(gender,age,home address,contact information)was recorded,and the clinical data of the two groups were statistically analyzed.A prospective parallel controlled study was used.According to the guiding principles for clinical research of new traditional Chinese medicine,the observation table is formulated according to the common clinical symptoms of patients,and the syndrome score is carried out according to the clinical symptoms.The normal group and the observation group were treated with normal observation and basic treatment for twenty-one days.The observation group counted the symptom scores at the time of enrollment and after treatment.The two groups were given:(1)the ACR and blood creatinine of the two groups were measured by automatic biochemical analyzer;(2)The expressions of SOD,MDA and SIRT1 in serum of the two groups before and after treatment were detected by ELISA.Result:The average age of the observation group was(46.80±7.55)years,and that of the normal group was(45.10±5.89)years.The average course of disease in the observation group was(1.66±0.56)years.Statistical analysis showed that there was no significant difference in age between the two groups(P>0.05).Creatinine value and symptom score can have an impact on TCM syndrome types,but age,gender and diet have little impact on TCM syndrome types.Through linear regression analysis,it is concluded that drug exposure time is a direct factor affecting the course of disease.Symptom score factors were positively correlated with course of disease,creatinine and MDA,and negatively correlated with SIRT1 expression.The disease degree of patients with drug-induced renal injury was mainly mild(sixty-five percent),The most common TCM syndromes are deficiency of spleen and kidney,damp heat accumulation syndrome(ninety percent respectively),the second is liver qi stagnation syndrome(seventy-five percent),blood stasis block syndrome(thirty percent),dampness and turbidity accumulation syndrome(ten percent).Deficiency syndrome(deficiency of both spleen and kidney)is often mixed with deficiency syndrome(deficiency of both spleen and kidney),but there are twenty cases(one hundred percent)related to dampness evil.To observe whether SIRT1/SOD/MDA is involved in the pathogenesis of drug-induced renal injury patients,we compared the indexes of the observation group and the normal group before treatment: it was found that MDA in the observation group was higher than that in the normal group,SOD was lower than that in the normal group,and the level of SIRT1 was lower than that in the normal group.The levels of ACR and creatinine in the observation group were higher than those in the normal group.Before treatment,the indexes of the two groups were statistically significant(P<0.05).Observing the relationship between the development of the disease and SIRT1/SOD/MDA,we compared the indexes of the two groups after treatment and the observation group before and after treatment(because the normal group was only used as the control without drug intervention): it was found that the level of MDA decreased after treatment,close to the normal group,compared with the normal group(P>0.05).The other indexes were still statistically significant compared with the normal group(P<0.05).The comparison of creatinine,ACR,oxidation indexes and other indexes in the observation group before and after treatment.The SIRT1 level in the observation group also increased with the reduction of symptoms and the improvement of various indexes before and after treatment.The comparison of various indexes before and after treatment was statistically significant(P<0.05).Pearson linear correlation analysis showed that there was a strong positive correlation between SIRT1 and SOD,and a strong negative correlation between SIRT1 and MDA,SOD and MDA;After treatment,there was a strong positive correlation between SIRT1 and SOD,and a weak negative correlation between SIRT1 and MDA,SOD and MDA.Conclusion:The common clinical syndrome types of DKI patients are damp heat accumulation,deficiency of spleen and kidney,stagnation of liver qi and blood stasis.Symptom score and creatinine can affect TCM syndrome types,while other factors have little effect on TCM syndrome types.It shows that the factors affecting the TCM syndrome type mainly come from the disease itself,and the lifestyle has little effect on the TCM syndrome type of DKI.The biological analysis of the serum of patients with DKI also suggests that oxidative stress is involved in the pathogenesis of DKI,and there is a strong correlation between oxidative stress and laboratory indexes such as serum creatinine and acr.The changes of laboratory indexes(the decrease of serum creatinine and ACR)will affect the changes of oxidative stress indexes(the increase of SOD/SIRT1 and the decrease of MDA).Linear regression showed that the course of DKI was affected by drug exposure time.The longer the drug exposure time,the more likely DKI occurred.The symptom score of patients is greatly affected by the course of disease,creatinine,MDA and SIRT1,that is,the longer the course of disease,the more serious the condition,the higher the symptom score and the more obvious oxidative stress in the body.Pearson linear correlation analysis showed that SIRT1 was correlated with SOD and MDA,indicating that the occurrence of drug-induced renal injury was closely related to oxidative stress.We draw a preliminary conclusion: SIRT1/SOD/MDA is involved in the pathogenesis of DKI,and the expression of SIRT1/SOD/MDA is closely related to DKI.The improvement of oxidative stress can participate in the process of renal repair.The up regulation of SIRT1 level will be accompanied by the changes of SOD and MDA and the reduction of renal injury. |
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