| BackgroundNowadays,hypertension is a chronic disease that affects nearly a third of adults worldwide.The number of hypertensive patients has reached 300 million in China.Therefore,hypertension has become one of the main influencing factors for the consumption of social medical resources,bringing increasingly serious economic burden to the country and the family.Target organ damage such as heart,brain and kidney caused by hypertension is a particular risk that cannot be ignored in patients with hypertension,among which the incidence of renal damage caused by hypertension is as high as 18%,second only to diabetic nephropathy and glomerulonephritis.In Chinese medicine treatment of hypertension,Professor Liu Duzhou major in Treatise on Febrile Disease founded Sancao Decoction(SCD),whose clinical curative effect is distinct.SCD is made of Peony and Licorice Decoction plus Gentian,Leonurus japonicus and Prunella vulgaris.It has the functions of clearing liver and purging fire,nourishing Yin and softening liver,promoting blood circulation and promoting water circulation.Our previous study has found that Sancao Decoction can significantly lower blood pressure through anti-inflammatory effect.Therefore,this study intends to explore whether it can further improve renal damage caused by hypertension through anti-inflammatory effect on this basis.Objective1.To observe the improvement of SCD on blood pressure,renal function,urine biochemical indexes and renal histomorphological changes in spontaneously hypertensive rats.2.By detecting the expression of Toll-like receptor 4(TLR4),Nuclear transcription factor-κB(NF-κB)，Nod-like receptor hot protein domain related protein 3(NLRP3)and other proteins in kidney tissues,the influence of SCD on the expression of TLR4/NF-κB/NLRP3 signaling pathway was explored,so as to identify the improvement of SCD in hypertensive renal damage and its possible mechanism.MethodSelecting 20 yearling 24 male Spontaneously Hypertensive Rats(SHR),which were randomly divided into model group,captopril group(CPT)(30 mg/kg),SCD group(SCD)(10.6 g/kg),8 male WKY rats of the same age were used as the normal control group(WKY).The blood pressure of the rats in each group was measured at week 4,week 8 and week 12,respectively.After 12 weeks of administration,the urine of the rats in each group was collected for 24 hours.Indexes such as urinary microalbumin(mAlb),urinary creatinine(UCRE)and urinary protein(UP)were detected by biochemical method.Blood was taken from the abdominal aorta affer the experiment,and serum was taken to detect the renal function indexes of rats in each group by biochemical method,such as blood creatinine(SCR),urea nitrogen(BUN)and blood uric acid(UA).Hematoxylin-Eeosin(HE)staining was used to observe the pathological changes of renal tissue,and Masson staining was used to evaluate the degree of renal fibrosis.Immunohistochemistry(IHC)was used to detect the expression of TLR4,MyD88,NF-κB,NLRP3 and IL-1β.Expression levels of monocyte chemokine 1(MCP-1)and IL-1β were determined by Enzyme-linked immunosorbent assay(ELISA).Expressions of TLR4,NF-κB and NLRP3 were detected by Western blot,Finally.SPSS 20.0 statistical software was used for data analysis.Results1.Changes of blood pressure and heart rate of rats in each group:before administration,Systolic Blood Pressure(SBP),Diastolic Blood Pressure(DBP)and Mean Arterial Pressure(MAP)in SHR group,CPT group and SCD group showed no significant difference(P>0.05),and Blood Pressure values of the above three groups were significantly higher than that of WKY group(P<0.01).After 4 weeks,8 weeks and 12 weeks after administration,systolic,diastolic and mean arterial pressures in the SHR group were significantly higher than those in the WKY group(P<0.01),while systolic,diastolic and mean arterial pressures in the CPT group and the SCD group were significantly lower than those in the SHR group(P<0.01).During the whole treatment process,compared with WKY group,the heart rate of SHR group,CPT group and SCD group was significantly higher,with statistically significant difference(P<0.01),while the heart rate of SHR group,CPT group and SCD group showed no statistically significant difference(P>0.05).2.Changes of urine biochemical indexes of rats in each group:after treatment,compared with the WKY group,the urinary microalbumin and urinary protein in the SHR group were significantly increased(P<0.01),the urinary creatinine was significantly decreased(P<0.01),the urine volume was significantly decreased(P<0.01),and the 24-hour urine albumin quantitative and urine protein quantitative were significantly increased(P<0.01).Compared with group SHR rats,SCD and CPT group rats urine trace albumin and urinary protein were significantly lower(P<0.01),and the urine creatinine concentrations in two groups weresignificantly increased(P<0.01),amount of urine SCD group significantly increased(P<0.01),whereas CPT group had no significant increased urine output(P>0.05),SCD group and 24 hours urinary albumin quantitative CPT group were significantly lower(P<0.01),but the two groups of 24 hours urinary protein quantitative no significant change(P>0.05).3.Changes of renal weight,body mass and renal index of rats in each group:after treatment,compared with the WKY group,the total weight of both kidneys in the SHR group was not significantly changed(P>0.05),but the body mass of the SHR group was significantly decreased(P<0.01),and the renal index in the SHR group was significantly increased(P<0.01).Compared with SHR group,SCD group and CPT group had no significant change in body mass(P>0.05),CPT group had significantly lower renal index(P<0.01),but SCD group had no significant decrease in renal index(P>0.05).4.Changes in renal function of rats in each group:after treatment,BUN,SCR and UA in SHR were significantly increased compared with WKY group(P<0.01).Compared with SHR group,urea nitrogen,blood creatinine and blood uric acid were significantly decreased in SCD group and CPT group,with statistically significant differences(P<0.01).5.Changes of renal histopathology of rats in each group:(1)HE staining results showed that,compared with the WKY group,some of the glomeruli in the SHR group were severely atrophic,with severe inflammatory infiltration around the renal arteries,obvious hyperplasia of mesangial matrix,and dilatation of renal tubules.Compared with the model group,SCD group and CPT group had less glomerular atrophy and no dilatation of renal tubules.(2)Masson staining results showed that compared with WKY group,collagen fiber deposition in SHR group was significantly increased,while collagen fiber expression in SCD group and CPT group was significantly decreased.6.Effect of SCD on TLR4/NF-κB/NLRP3 signaling pathway and expression of related inflammatory factors:IHC and Western blot results showed that compared with WKY group,TLR4,MyD88,NF-κB,NLRP3 and IL-1β expression levels in SHR group were significantly increased(P<0.01).Compared with SHR group,TLR4,MyD88,NF-κB,NLRP3 and IL-lβexpression levels in CPT group and SCD group were significantly decreased(P<0.01).The ELISA results showed that the expression levels of MCP-1 and IL-1βin SHR group were significantly higher than that in WKY group(P<0.01).Compared with SHR group,MCP-1 and IL-1β expression in CPT group and SCD group were significantly decreased(P<0.01).Conclusions1.Sancao Decoction can effectively improve renal damage caused by hypertension,which is mainly achieved by lowering blood pressure,improving biochemical abnormalities in urine,reducing the levels of blood creatinine,urea nitrogen and blood uric acid,and improving the pathological morphology of kidney tissues.2.Sancao Decoction can ihibit renal inflammation by regulating the TLR4/NF-B/NLRP3 signaling pathway,thereby improving renal damage caused by hypertension. |
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