Study On The Therapeutic Characteristics And Mechanism Of Diabetic Nephropathy By "treating From The Perspective Of Wind

BackgroundDiabetic kidney disease(DKD)is a chronic disease caused by diabetes.Due to its rapidly increasing incidence,poor prognosis,and heavy economic burden,DKD has become one of the important diseases threatening human health.Seeking treatment methods including Traditional Chinese medicine is the research hotspot of DKD."Wind-based treatment" is a new idea for DKD treatment proposed by Professor Zhao Jinxi,which has a good clinical effect and academic characteristics.This paper systematically summarizes the academic origin,medication experience,and clinical characteristics of "Wind-based treatment",and on this basis,it is of great significance to study the specific mechanism of DKD using animal and cell experiments.Objective1.Systematically summarize the academic origin and medication experience of "Wind-based treatment",and explore the clinical characteristics of "Wind-based treatment".2.From podocyte autophagy and podocyte protection,explore the mechanism of "Wind-based treatment" DKD on autophagic vesicle formation and degradation and further explore the scientific connotation of "Wind-based treatment" in DKD.Methods1.Clinical studiesThis study mainly adopts the method of combining qualitative research and quantitative analysis,including three aspects:sorting out the treatment ideas of "Wind-based treatment",studying the medication experience of "Wind-based treatment" and analyzing the advantages and characteristics of it.In study l,in the part of sorting out the treatment ideas of "Wind-based treatment",semistructured expert interviews and literature coding were used to conduct a qualitative analysis of the treatment ideas of "Wind-based treatment" by using information coding statistical technology,and the theoretical origin,treatment methods,advantages and characteristics of"Wind-based treatment" were summarized.In Study 2,the "Wind-based treatment" drug use empirical research gathering the Consilia of professor Zhao Jinxi and professor Lv Renhe on the treatment of DKD.Establishing a database of basis,and using correlation analysis,cluster analysis,centrality analysis of prescription data mining technology to analysis of drug law of "Wind-based treatment" by comparing the Consilia of professor Zhao Jinxi and professor Lv Renhe.This research lay a foundation for"Wind-based treatment’ basic medicine.In study 3,the clinical characteristics of "Wind-based treatment" were analyzed.The visual simulation scoring method was used to evaluate the subjective symptoms of DKD patients and analyze the clinical characteristics of "treating from wind" DKD.2.Experimental studiesThis study includes two aspects:an animal experimental study of DKD "Wind-based treatment"based on podocyte autophagy and a cell experimental study.In study 1,the animal experimental study of DKD "Wind-based treatment" of podocytes autophagy,db/db mice of spontaneous type 2 diabetes mellitus were used as experimental animals,and the DKD basic formula of "Wind-based treatment","Supplementing Qi,dispelling wind and clearing collaterals" and "dispelling wind and clearing collaterals" obtained by prescription data mining was used as experimental drugs.The autophagy promoter rapamycin,trehalose,and autophagy inhibitor chloroquine and 3MA were used as control.To analyze the improvement effect of "Wind-based treatment" on urinary ACR,renal pathological changes and podocyte autophagy-related indexes of db/db mice.In study 2,in the experimental study of DKD cell "Wind-based treatment" based on podocyte autophagy,in order to better reflect the clinical situation,two modeling methods of highglucose culture medium and serum of DKD patients with massive proteinuria were used for intervention.The autophagy promoter rapamycin,trehalose,and autophagy inhibitor chloroquine,3MA were compared with the "Wind-based treatment" basic prescription "Yi Qi Qu Feng Tong Luo formula" and "Qu Feng Tong Luo formula".The protective effect of DKD"Wind-based treatment" on podocytes and its specific effect on autophagy was investigated in mouse podocytes.Results1.Relevant clinical studies(1)Review results of "Wind-based treatment" in DKD:six related researchers were interviewed,and 16 related articles were included.Three-level coding was conducted for interviews and literature coding,which were summarized into four aspects:theoretical origin,pathogenesis understanding,medication methods and curative effect advantages.(1)It is found that the theory of "Wind-based treatment" originates from Professor Lu Renhe’s theory of "micro-lump syndrome" and is influenced by the treatment ideas of Professors Huang Wenzheng and Zhao Shaoqin.Second,the idea of "Wind-based treatment" emphasizes that the core pathogenesis of DKD is "latent wind in kidney",including internal wind and external wind.The treatment for the pathogenesis of "latent wind in kidney" runs through DKD,regardless of whether there are typical manifestations of wind evil.Third,In view of the pathogenesis of DKD "latent wind in kidney",drugs such as dispelling wind and dispersing evil,dispelling wind and dehumidifying,calming liver and calming wind are often used for treatment,which need to be combined with tonifying qi,blood circulation and collaterals treatment in a specific application.Forth,"Windbased treatment" can mainly play a role in improving renal function and improving the overall curative effect.At the same time diabetic retinopathy,arteriosclerosis and other complications have a certain treatment effect.(2)Experience results of "Wind-based treatment" in DKD:331 patients cured by Professor Zhao Jinxi were included in the study,with 518 clinical times.74 patients cured by Professor Lu Renhe were included in the study,with 171 clinical times.Data mining results show that Professor Lu Renhe attaches great importance to the treatment of deficiency,blood circulation,clearing heat,and so on.Drug centrality analysis showed that the core drugs included Dan Shen,Chuan Xiong,Dan Pi,Chi Shao,and so on.Professor Zhao Jinxi attaches great importance to nourishing qi and blood,activating blood circulation,improving water infiltration and dispelling wind,and relieving the surface.Its clinical core drugs are Huang Qi,Dan Shen,Tu Fu Ling,Bi Xie,and so on.In order to deal with the pathogenesis of "latent wind in kidney",Professor Zhao Jinxi commonly used drugs,such as Bi Xie,Chuan Shan Long,Niu Bang Zi,Chan Tui,and so on.The results of cluster analysis showed that the common drug combinations of Professor Zhao Jinxi were Huang Qi,Dan Shen,Tu Fu Ling,Bi Xie,Chuan Shan Long,and so on.By comparing the medication experience of the two experts,it was found that Professor Zhao Jinxi "Wind-based treatment"more often uses drugs such as dispelling wind and relieving surface,dispelling wind and dampness.In terms of drug combination and symptom-drug correlation analysis,the two experts have many similar medication methods,suggesting that Professor Zhao Jinxi’s "Wind-based treatment" may be related to Professor Lu Renhe’s theory of "micro-lump syndrome".Based on the data mining results,the basic DKD prescriptions of"Wind-based treatment" were formed as Yiqi Qu Feng Tong Luo formula and the Qu Feng Tong Luo formula.(3)Results of the study on the characteristics of "Wind-based treatment" in DKD:73 patients with mid-stage DKD in Professor Zhao Jinxi’s outpatient department were included in this study,with 156 clinical times.46 patients with advanced DKD had 96 clinical times.It was found that the wind-related symptoms such as urine foam,facial edema,dizziness,and skin pruritus in patients with middle and late DKD were significantly improved after treatment.The overall health status and quality of life of patients were also significantly improved.The improvement of patients with late DKD was better than that of patients with middle DKD.2.Experimental studies(1)The experimental results of "Wind-based treatment":after 8 weeks of intervention,the ACR of mice in the model group was significantly higher than that in the blank group,indicating that the DKD model was successfully established.However,the ACR of mice was significantly decreased after the treatment with the basic DKD prescriptions of Yi Qi Qu Feng Tong Luo and Qu Feng Tong Luo prescription.HE staining and PAS staining showed mesangial hyperplasia and basement membrane thickening in the model group.The renal pathological injury was less in Yi Qi Qu Feng Tong Luo group And Qu Feng Tong Luo group.Under the electron microscope,the number of autophagic vesicles in the glomerular podocytes of Yi Qi Qu Feng Tong Luo group and Qu Feng Tong Luo group was significantly increased compared with the model group.At the same time,the basic formula of ""Wind-based treatment" can significantly increase the expression of nephrin and podocin in mouse kidney,increase the content of autophagic vesicle formation related proteins such as ATG5,ATG7,Beclin1,LC3II/LC3I,and significantly inhibit the increase of P62 in the model group.(2)The results of the "Wind-based treatment" cell experiment of DKD:the two methods of serum modeling of DKD patients with 30 mmol/L glucose and massive proteinuria can lead to the inhibition of MPC5 cell proliferation,cytoskeleton damage and the decline of nephrin index.After the treatment with Yi Qi Qu Feng Tong Luo formula and Qu Feng Tong Luo formula,the structural damage of MPC5 cells improved and nephrin index recovered.On the other hand,the basic formula of "Wind-based treatment" could significantly increase ovalbumin staining of cells,significantly increase the expression of ATG5,ATG7,Beclinl and other indicators,and significantly inhibit the increase of P62 in the model group.The double fluorescent labeling of LC3 protein also suggested that both digested and undigested autophagic vesicles in cells were significantly increased after treatment with the basic recipe of "Wind-based treatment".The level of cathepsin L in "Wind-based treatment" groups also increased significantly.Conclutions1.The idea of "Wind-based treatment" inherits from Professor Lu Renhe’s theory of "microlump syndrome" and attaches great importance to the basic pathogenesis of "kidney collaboration Fufeng".Commonly used anti-wind drugs include wind-dispelling drugs,antiwind and unblock circulation drugs,and so on.The clinical treatment attaches importance to the combination of dispelling wind and nourishing qi and activating blood.2."Wind-based treatment" commonly used drugs include Huang Qi,Dan Shen,Tu Fu Ling,Bi Xie,Chuan Shan Long,and so on.In order to deal with the pathogenesis of "latent wind in kidney",Professor Zhao Jinxi commonly used drugs,such as Bi Xie,Chuan Shan Long,Niu Bang Zi,Chan Tui,and so on.Based on the data mining results,the basic DKD prescriptions of "Wind-based treatment" were formed as Yiqi Qu Feng Tong Luo formula and the Qu Feng Tong Luo formula.Clinical observation shows that "Wind-based treatment"may have characteristics in improving the overall quality of life and relieving the symptoms related to wind evil.3."Wind-based treatment" basic formulas--Yiqi Qu Feng Tong Luo prescription and Qu Feng Tong Luo prescription can improve the urinary protein level of db/db mice and reduce the pathological damage of glomeruli.At the same time,it can increase the protein expression levels of nephrin and podocin in podocytes,which has a protective effect on kidney cells.In addition,the basic formula of "Wind-based treatment" can also improve ATG5,ATG7,Beclinl,LC3II/LC3I,and other indicators in mouse kidneys,suggesting that it may promote the formation stage of autophagic vesicles.4.The results of the cell study showed that the basic formula of "Wind-based treatment" had a protective effect on the structure and function of podocytes,and could improve the ATG5,ATG7,Beclinl,LC3Ⅱ/LC3Ⅰ,cathepsin L and other indicators of podocyte injury model,and might promote the formation of autophagic vesicles and the digestion process.The protective effect of the DKD basic formula on podocytes may be related to the improvement of autophagy.