| BackgroundDiabetic cognitive impairment(DCI)is a serious central nervous system complication induced by diabetes.However,the prevention and treatment methods for DCI are currently limited.Therefore,it is necessary to actively explore relative high-risk factors,which are crucial for early screening and prevention of DCI.At present,a growing body of research on cognitive impairment is now focusing on the heart-brain connection,and the left ventricular hypertrophy is considered as an independent factor for cognitive impairment.However,the correlation of left ventricular hypertrophy with cognitive impairment in the context of diabetes have not been reported in the literature yet.Traditional Chinese medicine(TCM)has certain advantages in the treatment of diabetic complications.Yi-Xiao Formula,a TCM herbal compound based on the "micro-zhengjia" theory,is created for the treatment of diabetic complications.This study further explored the mechanism of Yi-Xiao Formula on cerebral complications of diabetes,in order to provide a theoretical basis for clinical practice and new drug development.Objective1.To observe the correlation of left ventricular hypertrophy with the impairment in overall and different domains of cognitive function in the context of diabetes,to provide a basis for early screening and prevention of cognitive impairment in diabetes.2.To observe the distribution of TCM syndrome elements in patients with diabetes mellitus with and without left ventricular hypertrophy,and to explore the correlation between different syndrome elements and left ventricular structural changes and cognitive function,than establish the TCM syndrome relationship of diabetes mellitus with cardio and cerebral complications.3.Based on the previous study results,to observe the effect of Yi-Xiao Formula intervention on the improvement of hippocampal pathological damage in KKAy mice,and to observe the effect of Yi-Xiao Formula intervention on TGF-β/Smads signaling pathway in hippocampus.To explore the intervention targets of Yi-Xiao Formula in the treatment of central nervous system complications of diabetes based on TGF-β/Smads pathway.Methods1.Using the method of cross-sectional research,the inclusion and exclusion criteria were formulated according to the diagnostic criteria of diabetes mellitus and left ventricular hypertrophy.A total of 140 patients with or without left ventricular hypertrophy were included,70 patients each group.The general condition,medical history data,laboratory test data,echocardiographic test data,and cognitive-related scale test data such as Mini-Mental State(MMSE)and Montreal Cognitive Assessment(MoCA)were collected.The TCM syndromes were determined according to the information of the "Four Diagnoses" in TCM,and the Case Report Form(CRF)was filled out.The data of each group were entered into SPSS 22.0 for statistical analysis.2.The 12-week-old spontaneous type 2 diabetes KKAy mice were selected as the experimental group,and the same-week-old C57BL/6J mice were selected as the control group.After 2 weeks of adaptive feeding,KKAy mice with fasting blood glucose>13.9mmol/L were considered as diabetic mice and were included in the following experiments.Before grouping,6 KKAy mice and 6 C57BL/6J mice were randomly sacrificed,and the their brain tissues were collected for baseline pathological evaluation.The remaining KKAy mice were randomly divided into the model group,the Yi-Xiao Formula group and the losartan group,with 18 mice in each group.18 C57BL/6J mice with normal level of blood glucose were selected as the control group.The Yi-Xiao Formula group was given Yi-Xiao Formula 7.7g/(kg·d)by gavage,while the losartan group was given losartan 10mg/(kg·d)by gavage.The mice in the control group and model group were given an equal dose of distilled water daily by gavage.The gavage intervention lasted 12 weeks.At the 4th,8th,and 12th week of intervention,6 mice in each of the four groups were randomly selected and sacrificed after anesthesia,and brain tissues were collected for HE staining,Nissl staining and pathological analysis.The relative protein level of TGF-β1,p-Smad2,p-Smad3,Smad7 in hippocampus were detected by Western blot.The relative mRNA level of TGF-β1 and Smad7 in hippocampus were analyzed by RT-qPCR.Results1.The results of the cross-sectional research showed that at baseline,the weight,BMI,HbA1c level and the prevalence of hypertension in the diabetes with LVH group were significantly higher than those in the non-LVH group(P<0.05).The LVMI,LVIDd,LVPWd and I VST level in the diabetes with LVH group were significantly higher than those in the non-LVH group(P<0.01),and there was no significant difference between two groups in cardiac function data.In terms of cognitive function,the MoCA total score,visuospatial and executive ability,language and delayed recall and other cognitive domains scores of the diabetes with LVH group were significantly lower than those in the non-LVH group(P<0.05).The results of correlation analysis showed that LVMI,LVPWd,IVST and MoCA total scores,visual space and executive ability,language and delayed recall and other cognitive domains were significantly negatively correlated(P<0.01).Logistic regression analysis showed that after adjusting for confounding factors,LVH was not associated with the diagnosis of MCI,while LVMI,LVIDd,and LVPWd were independently associated with the diagnosis of MCI.Each unit of increase in the above indicators increased the risk of diagnosing MCI by 2.6%(OR:1.026,P<0.01),15%(OR:1.150,P<0.01)and 58%(OR:1.580,P<0.01)respectively.2.The results of the investigation of TCM syndrome elements showed that the deficiency syndrome elements of the two groups were mostly qi deficiency and yin deficiency,and the solid syndrome elements were mainly dryness and heat,phlegm-dampness,qi stagnation and blood stasis.The most common TCM syndrome elements of disease site are kidney,lung,heart and liver.The distribution of the overall TCM syndrome elements basically conforms to the pathogenesis of Xiaoke disease,which is characterized by"deficiency of both qi and yin,and excessive dryness and heat".Comparison between groups showed that the frequency of phlegm-dampness,blood stasis,phlegm-dampness-blood stasis complex syndrome elements and the frequency of heart-qi deficiency syndrome elements in the diabetes with LVH group were significantly higher than those in the non-LVH group(P<0.05).Analysis of the relationship between TCM syndrome elements and LVMI showed that the overall LVMI level of patients with phlegm-damp,blood stasis,phlegm-damp-blood stasis complex TCM syndrome elements and heart-qi deficiency TCM syndrome elements was significantly higher than that of patients without such TCM syndrome elements(P<0.01).Further analyzing the distribution of TCM syndrome elements in patients diagnosed with MCI in the two groups,Phlegm-dampness and blood stasis were more common in patients with diabetes diagnosed with MCI,and the disease location involved heart and kidney.The intra-group comparison showed that the frequency of phlegm-damp,blood stasis,phlegm-damp-blood stasis complex TCM syndrome elements and heart-qi deficiency TCM syndrome elements in MCI patients in the diabetes with LVH group were significantly higher than those in non-MCI patients(P<0.01).The distribution of TCM syndromes elements in diabetes with non-LVH patients with MCI was basically the same as that in non-MCI patients.Analysis of the relationship between TCM syndrome elements and MoCA score showed that the overall MoCA score of patients with phlegm-damp,phlegm-damp-blood stasis complex syndrome elements was significantly lower than that of patients without such TCM syndrome elements(P<0.01).3.The results of experiments showed that in terms of glucose metabolism,the FBG level in the Yi-Xiao Formula group was significantly lower than that in the model group from the 8th week of intervention(P<0.01).In terms of the pathology of hippocampal HE staining,the neurons in the CA1 and CA3 regions of the hippocampus of the mice in the model group were loosely arranged since the 4th week of intervention,the cell membrane and nucleolus were blurred,and some pyknotic and hyperstained apoptotic cells and necrotic cells were observed.And with the development of the disease,it can develop into an area of necrotic cells,and the above-mentioned pathological changes can be alleviated to varying degrees after the intervention of Yi-Xiao Formula.In terms of Nissel staining pathology,the number of neurons in the hippocampal CA1 and CA3 regions of the Yi-Xiao Formula group was significantly increased compared with the model group(P<0.05).In terms of protein expression levels in hippocampal tissue,the levels of TGF-β1,p-Smad2/Smad2,p-Smad3/Smad3 and APP in the Yi-Xiao Formula group were significantly lower than those in the model group(P<0.05),while the level of Smad7 in the Yi-Xiao Formula group was significantly higher than those in the model group(P<0.01).In terms of mRNA levels in hippocampus,the mRNA levels of TGF-β1 and APP in the Yi-Xiao Formula group were significantly lower than those in model group(P<0.01),and the mRNA level of Smad7 in the Yi-Xiao Formula group was significantly higher than those in model group(P<0.01).Conclusions1.In this study,a cross-sectional research was conducted and found that after adjusting for related confounding factors,increased levels of LVMI,LVPWd,and LVIDd in diabetic patients could increase the risk of MCI.2.Diabetic patients with LVH and patients diagnosed with MCI in this group are characterized by phlegm-dampness,blood stasis,phlegm-dampness and blood stasis.Phlegm-damp syndrome elements dominated the correlation with cognitive dysfunction,followed by blood stasis syndrome elements.It preliminarily verified the view that"micro-zhengjia" widely exists in the complications of diabetes mellitus.3.Yi-Xiao Formula could inhibit the expression of genes and proteins of the TGF-β/Smads signaling pathway in the hippocampus,relieve hippocampal neuropathy,and APP protein expression.Combined with previous studies,it is suggested that diabetic cardiomyopathy and hippocampal neuropathy are jointly affected by TGF-β/Smads signaling pathway,such mechanism may be a microscopic concrete manifestation of "micro-zhengjia"as the common pathogenesis of complication of diabetes mellitus. |
PDF Full Text Request