San Qi Oral Solution, The Chinese Medicine Compound Recipe Of Benefiting Vital Energy And Promoting Blood Flow: Clinical Observation And Experimental Study Of Its Renal Protective Effects On The Patients Suffered From Diabetes Nephropathy

BackgroundRecently,the prevalence rate of diabetes is rising unceasingly in our country.Along with insulin widespread application,the mortality rate of Diabetes' acute complication,such as ketoacidosis,are obviously dropped,and the chronic complication such as diabetic nephropathy has become one of most common complications with diabetic life extension.In clinical practice,diabetes nephrosis is often divided into 3 stages,namely:morning stage,intermediate stage and advanced stage.In different stage,the emphasis of treatment is also different.Professor Yang consider that the Deficiency of Qi and the Blood Stasis is the mainly etiopathogenesis of diabetic nephropathy,especially when disease is morning or intermediate stage,so the therapeutic rule- "benefiting vital energy and promoting blood flow" can applied in the treatment of diabetic nephropathy;when their stage are morning or intermediate,the effective treatment is the key of pathogenetic condition reverse.So we study the mechanism of action of SanQi Oral Solution to the diabetic nephropathy by the methods of pharmacodynamics and morphology,in order to provide the substantial theory and plenitudinous scientic evidence for SanQi Oral Solution which is composed according to benefiting vital energy and promoting blood flow method cure diabetic nephropathy.Objective.To explore the effect and mechanism of SanQi Oral Solution in the defence and treatment of morning diabetic nephropathy,and found SanQi Oral Solution' s function superiority stage.Methods:1).Clinical study:This part of the study is controlled.According to degree of microalbuminuria and general condition,58 patients suffered from morning stage diabetic nephropathy were divided into treatment and control group. There was no statistically significant difference between two groups in sex, age,severity of the disease and laboratory items.The patients of the treatment group received management of SanQi Oral Solution while the patients of the control group took the drug of Lotensin.The course of treatment lasted for 12 weeks,during which the changes of clinical findings and lab index were monitored and recorded,including renal function,microalbuminuria,blood fat, blood pressure,blood glucose,thrombomodulin and clinical symptoms and signs. The differences between the symptom index before and after treatment were then calculated and tested,as well as the microalbuminuria of 24 hours and thrombomodulin.2).Experimental Study:(1) PartⅠ.Study of effects of the SanQi oral solution on the expression of AGEs and its acceptor,as well as regulation factor42 SD male adult rats of specified-pathogen free were injected with streptozotocin(STZ) intraperitoneally to prepare for Diabetes Mellitus animal model and then randomly divided into 3 groups including model group(M group),SanQi Oral Solution group(S group) after diabetes was induced successfully.Another group was set for experiment control,consisted of normal SD rats.After diabetes was induced successfully,the rats of diabetes animal model were continually raised with high fat food every other day for another 4 weeks in order to induce the diabetes nephropathy in them.After 6 weeks of drugs treatments or placebo managements,all the animals were put to death and their kidneys were removed for the pathological study.6 renal samples were randomly obtained from the rats of group C,M,and T in PartⅡstudy,respectively.Expression of AGEs/RAGE,PAI-1 and TGF-β1 were detected by immunohistochemistry.The stained area,area density,and photodensity were then observed and recorded.(2) PartⅡ.Study of effects of the SanQi oral solution on the DN rats' podocyte as well as the expression of the PCX and Nephrin.6 renal samples were randomly obtained from the rats of group C,M,and T in PartⅠstudy,respectively.Expression of PCX and Nephrin were detected by immunohistochemistry.The methord is the same as PartⅠ.Results:1).Clinical study.(a).there' s no statistical significant difference in the general condition of the patients,the urinary protein of 24 hours,and renal function before treatment,No statistical significance was found in the difference of blood pressure,values of blood glucose during the course of treatment.(b).As to the patients of T group,the symptomatic scores,TM and microalbuminuria of 24 hours after treatment decreased obviously compared to that before the treatment,with statistical significance(P＜0.01).(c).There were no statistical significance found in the difference of changes of urinary protein after treatment between the T group and L group (P＞0.05).However,the changes of symptomatic scores in T group was higher than that of L group after treatment with statistical significance(P＜0.01).2) Experimental Study:(1).Study of effects of the SanQi oral solution on the expression of AGEs and its acceptor,as well as regulation factor.a.microalbuminuria of 24 hours,urinary protein in T,and M group increased with statistical significance,compared to that of C group(P＜0.01).Excretion of urinary protein in 24 hours in T group decreased statistically significantly, in contrast to that of M group(P＜0.01).b.Changes of values of blood urea nitrogen and serum creatinine.There' s no statistical significant difference demonstrated between C,T,and M groupc.The cholesterol total(TCh),low density lipoprotein-oholesterol (LDL-C)and glycerol total(TG) fluctuation.The mean values of TCh and TG in T,and M group increased statistically significantly,compared that of C group(P＜0.01).In T group,the values of TCh,LDL-C and TG decreased with statistical significance,compared to that of M group(P＜0.01).d.Expression of AGEs.The AGEs was located in the glomerulus,kidney tubules,capillaries,mesenteria and etc.And deposited at the kytoplasm and Extracellular Martix.Strong positive expression was found in group M. According to the outcomes of immunohistology,the values of area density and photodensity of positively stained area were less than that of M group with statistical significance(P＜0.01).e.Expression of RAGE.The expression of RAGE was located mainly in glomerulus.It' s more strongly expressed in group M than in C group.The value of area density and photodensity in M group positively stained area were higher than that of T group(P＜0.01).f.Expression of PAI-1.Expression of PAI-1 was located in renal tubule and collecting pipe but no expression located in glomerulus.It' s stronger positively expressed in M group than in C.The values of area density and photodensity of PAI-1 positively stained area in group T were lower than that of group M with statistical significance(p＜0.05).g.Expression of TGF-β1.Expression of TGF-β1.was located in the renal tubule and interstitium.It' s weakly expressed in the glomerulus.It' s lower for the value of area density and photodensity in group T than that of group M with statistical significance(P＜0.01).(2)Study of effects of the SanQi oral solution on the DN rats' podocyte as well as the expression of the PCX and Nephrin.a.Glomerulus foot cell count:Compared with the C group,M group glomerular podocyte number decreased obviously.(P＜0.05).T Group glomerular podocyte number also dropped,but cells is more than the M group,the results of statistical analysis different significantly(P＜0.05)b.Expression of PCX:NC Group PCX-positive signal can be seen along the glomerular vessel wall distribution running continuously;M Group PCX distribute not only uneven signals,but also incontinuous;Compared with the M group,T PCX-positive group signals distributed continuously and uniformly.Quantitative analysis revealed that,M Group PCX mean fluorescence density were lower than C,T group (48.5%lower,respectively,30.1%)(respectively P＜0.001,P＜0.05).c.Expression of NephrinM Group glomerular nephrin protein expression was significantly lower than C group(P＜0.05);T Group glomerular nephrin protein expression was significantly higher than M group(P＜0.05);T group compared with C group, the expression of nephrin protein has no significant difference(P＞0.05). d.Analysis of glomerular podocyte indicators with a 24-hour urine protein relevance:Analysis of results showed that the negative correlation exist among thd 24h UMA,expression of PCX and nephrin.(P＜0.01) Conclusion(1).SanQi Oral Solution can improve the patients' clinical symptoms and renal function,decrease the urine protein,and the level of TM,stable renal function,results suggest that SanQi Oral Solution may be used to treat DN as soon as Thrombomodulin increased.(2).The mechanism that SanQi Oral Solution ameliorated the structure and function damage of DN model rats may be due to inhibition of AGEs/RAGE,down-regulation of the expression of TGF-β1,increase of degeneration of PAI-1,reduction of extracellular matrix accumulation,and amelioration of renal pathological lesions.These therapeutic effects may improve renal hypertrophy,regulation of cells,ECM,cytokines,and growth factors resulting in delayed progress of DN.(3).SanQi oral liquid medicine can up-regulate kidney podocyte nephrin, podocyte-specific marker proteins of the protein expression PCX so as to improve podocyte damage,improve podocyte ultrastructure of the kidney to reduce urinary protein,delay DN process.