Efficacy And Mechanism Of Shenwu Huangxue Decoction In The Treatment Of End Stage Renal Disease With Cognitive Impairment Of Kidney Deficiency And Phlegm-blood Stasis Syndrome

Part Ⅰ1.Study of cognitive function and TCM syndrome differentiation of peritoneal dialysis patientsObjective:To systematically evaluate the incidence of cognitive impairment(CI)of peritoneal dialysis(PD)patients in the peritoneal dialysis center of Nephrology Department of Changshu Affiliated Hospital of Nanjing University of traditional Chinese medicine;to provide the basis for the establishment of TCM diagnosis and treatment plan for e end-stage renal disease with cognitive impairment(ESRD-CI).Methods:The cognitive function of PD patients in peritoneal dialysis center was measured by Montreal Cognitive Assessment Scale(MoCA V7.3);the four diagnostic information of traditional Chinese medicine was collected for the patients with CI and the distribution of each syndrome type was counted.Results:A total of 133 PD patients were enrolled in this study,there were 96 patients with MoCA score less than 26(CI),accounting for 72.2%of the total.Among them,the three cognitive domains with the highest proportion of impairment are:memory,attention,execution;the 96 esrd-ci patients were diagnosed with TCM syndrome differentiation.The main syndrome types were as follows:kidney essence deficiency with phlegm turbidity and blood stasis in 61 cases(63.5%),kidney essence deficiency with phlegm turbidity in 2 cases(2.1%),Qi and blood deficiency with phlegm turbidity and blood stasis in 19 cases(19.8%),Qi and blood deficiency with blood stasis in 5 cases(5.2%),Qi and blood deficiency in 6 cases(6.3%),liver and kidney yin deficiency in 3 cases(3.1%).Conclusion:The number of PD patients with CI in our hospital peritoneal dialysis center reached 96 cases,accounting for 72.2%of the total;the most common syndrome types of these patients were kidney essence deficiency with phlegm turbidity and blood stasis syndrome,accounting for 63.5%.2.Clinical evaluation of Shenwu huangxue Decoction in the treatment of peritoneal dialysis with cognitive impairmentObjective:To evaluate the effect of Shenwu huangxue Decoction on peritoneal dialysis patients with cognitive impairment.Methods:Sixty CI patients with kidney essence deficiency syndrome and phlegm stasis syndrome in our center were enrolled in a prospective cohort study with positive control.The control group was treated with regular peritoneal dialysis and routine medicine treatment,such as controlling blood pressure and correcting anemia.The treatment group was treated with Shenwu huangxue Decoction on the basis of the control group,and the course of treatment was three months.Results:A total of 57 patients completed the experimol group,the executive function(P<0.05)and the total scores(P<0.05)of the treatment group were significantly improved.There was no significant difference in oent,28 in the control group and 29 in the treatment group.Compared with the contrther domains between the two groups;compared with the control group,the serum IS of treatment group decreased significantly(P<0.01);there was no significant difference in the changes of BUN and SCr between the two groups after treatment.Conclusion:Shenwu Huangxue Decoction has good effect on peritoneal dialysis patients with CI whose syndrome type were kidney essence deficiency and phlegm stasis block,and can reduce the level of serum IS effectively.Part Ⅱ1.Effect of Shenwu huangxue Decoction on the mouse model of end-stage renal disease complicated with cognitive impairmentObjective:To study the therapeutic effect of Shenwu huangxue Decoction(SHD)on the mouse model of end-stage renal disease complicated with cognitive impairment(ESRD-CI),and to provide basic theoretical basis for clinical treatment of the disease.Methods:5/6 nephrectomy was used to establish a mouse model of ESRD,which was divided into sham operation group,model group,positive drug group,SHD high dose group and SHD low dose group.The mouse in each group were fed continuously,the water maze experiment(Morris)was carried out and the cognitive function of the mice in each group was detected,and the ESRD-CI mouse model was established.SHD high dose group and SHD low dose group were given 24 g/kg·d and 12 g/kg·d concentration,respectively.The sham operation group and the model group were given the same amount of normal saline,and the positive group was given donepezil 10 mg/kg.After 4 weeks of continuous gavage,the Morris water maze experiment was performed again to record the cognitive function of mice in each group,and the blood biochemical indexes and brain pathology of mice in each group were detected.Results:compared with the model group,the escape latency of SHD high-dose group and SHD low-dose group was significantly decreased,the difference was statistically significant(P<0.05).The original platform quadrant residence time and crossing times were significantly increased,the difference was statistically significant(P<0.05).Compared with the model group,the serum and brain tissue concentration of IS for SHD group were significantly decreased,the difference was statistically significant,and was concentration dependent(P<0.01).The brain concentration of MDA、IL-1β、TNF-α in high and low dose groups decreased significantly,difference was statistically significant(P<0.05),And the activity of SOD increased significantly,the differences were statistically significant too(P<0.01).After treatment,the pathological morphology of hippocampal tissue about the mouse in high-dose group and low-dose group were improved to varying degrees,especially in high-dose group.Conclusion:SHD may improve the cognitive function of ESRD-CI mice by reducing the concentration of IS,inhibiting the inflammatory reaction and the production of lipid peroxide and enhancing the scavenging effect of free radicals.2.Mechanism of Shenwu huangxue Decoction in the treatment of end stage renal disease with cognitive impairmentObjective:To investigate the mechanism of SHD in the treatment of ESRD-CI by immunohistochemistry,immunofluorescence,RT qPCR and Western blot.Methods:The expressions of NF-κB,MAPK,JNK,OAT1,BDNF,TrkB and AHR protein and mRNA in brain and kidney were detected by immunohistochemistry,immunofluorescence,Western blot(WB)and RT qPCR.Results:Compared with the model group,the number of NF-κB,MAPK,JNK and AhR positive cells in brain and kidney of SHD high-dose group and low-dose group was significantly decreased,while the number of OAT1 positive cells in brain and kidney and the one of BDNF,TrkB positive cells in hippocampus were significantly increased.Moreover,SHD high dose increased the expression of BDNF,TrkB protein and mRNA in brain,kidney and hippocampus of mice,and the expression of NF-κB,MAPK,JNK and AhR protein and mRNA in brain and kidney was the most.Conclusion:SHD can protect neurons by inhibiting IS mediated AhR/NF-KB/JNK signaling pathway,which may be achieved by increasing the activity of BDNF/TrkB pathway.3.Study on the material basis of Shenwu huangxue DecoctionObjective:To analyze and identify the chemical constituents in SHD by LC-MS,so as to lay a foundation for the study of the treatment for ESRD-CI by SHD.Methods:Through the comprehensive analysis of the retention time,relative molecular weight and other information of each component in the chromatogram,the effective components were determined by comparing with the reference substance or literature reports.Results:A total of 82 main chemical constituents were identified,including ginsenoside,emodin,aloe emodin,β-asarone,etc.Conclusion:The material basis of SHD is preliminarily clarified.