| 1.Objective To observe the clinical effect of Shendi granule,a Chinese traditional medicine with the effect of invigorating spleen,nourishing kidney and strengthening essence,on patients with chronic glomerulonephritis（CGN）with spleen and kidney deficiency syndrome and the intervention effect of podocyte injury.2.Methods48 CGN patients with spleen-kidney deficiency syndrome were randomly divided into Shendi granule group（24 cases）and Valsartan group（8 cases）,with a total of 40 cases completed（20 cases in each group）.Patients in Shendi granule group and Valsartan group were treated with Shendi granule and Valsartan capsule respectively for8 weeks.24 h urinary protein quantification（24h Upro）,urinary albumin/urinary creatinine ratio（urinary ACR）,urine podocyte marker protein PCX and Nephrin,renal function [blood creatinine（SCR）,blood urea nitrogen（BUN）,estimated glomerular filtration rate（EGFR）] before and after treatment in 2 groups were observed,and clinical disease efficacy and TCM syndrome efficacy were evaluated.In addition,the safety indexes of the two groups were also observed.Forty-eight CGN patients with spleen-kidney deficiency syndrome were randomly divided into Shendi granule group（24 cases）and Valsartan group（8 cases）,with a total of 40 cases completed（20 cases in each group）.Patients in Shendi granule group and Valsartan group were treated with Shendi granule and Valsartan capsule respectively for 8 weeks.24 h urinary protein quantification（24h Upro）,urinary albumin/urinary creatinine ratio（urinary ACR）,urine podocyte marker protein PCX and Nephrin,renal function [blood creatinine（SCR）,blood urea nitrogen（BUN）,estimated glomerular filtration rate（EGFR）] before and after treatment in 2 groups were observed,and clinical disease efficacy and TCM syndrome efficacy were evaluated.In addition,the safety indexes of the two groups were also observed.3.Results3.1 Comparison of clinical disease efficacy: the total effective rate of clinical disease efficacy in the Shendi granule group was 85%,and that in the Valsartan group was 50%.The total effective rate of clinical disease efficacy in the Shendi granule group was significantly higher than that in the Valsartan group,with statistical significance（P< 0.05）.3.2 Comparison of efficacy of TCM syndromes: the total effective rate of TCM syndromes in the Shendi granule group was 85%,and that in the Valsartan group was 60%.The total effective rate of TCM syndromes in the Shendi granule group was significantly higher than that in the Valsartan group,with statistical significance（P <0.05）.3.3 Comparison of TCM syndrome scores:there was no significant difference between the two groups before treatment（P > 0.05）;After 8 weeks of treatment,the TCM syndrome scores in Shendi granule group were significantly decreased（P < 0.05）,but not significantly in valsartan group（P BBB 0 0.05）.After 8 weeks of treatment,there was also significant difference in TCM syndrome scores between Shendi granule group and valsartan group（P < 0.05）.3.4 Comparison of 24 h Upro and urinary ACR: Before treatment,there were no significant differences in 24 h Upro and urinary ACR between the two groups（P >0.05）;At the 4th week of treatment,24 h Upro and urinary ACR of 2 groups were decreased compared with before treatment,but the difference was not statistically significant（P > 0.05）.At week 8,24 h UPRO and urinary ACR in 2 groups were significantly lower than those before treatment（P<0.05）.In addition,there were no significant differences in 24 h Upro and urinary ACR between the two groups at the 4th week of treatment（P > 0.05）,while the 24 h Upro and urinary ACR in Shendi Granules group at the 8th week of treatment were significantly lower than those in Valsartan group at the same period（P<0.05）.3.5 Comparison of renal function: There were no significant differences in renal function（SCR,BUN,EGFR）between the two groups before treatment,and SCR,BUN,EGFR after 8 weeks of treatment compared with that before treatment（P > 0.05）.3.6 Comparison of urinary PCX and Nephrin protein contents: Compared with the normal group,the urinary PCX and Nephrin contents in Shendi granule group and valsartan group were significantly increased,with statistical significance（P<0.05）.At the 8th week of treatment,the contents of urinary PCX and Nephrin in Shendi granule group and valsartan group were significantly decreased compared with those before treatment（P<0.05）.The contents of urinary PCX and Nephrin in Shendi granule group were significantly lower than those in valsartan group（P<0.05）.4.Conclusion1.Podocyte injury may be one of the important mechanisms of CGN proteinuria.2.Shendi granules,a traditional Chinese medicine with the effect of Jianpi-yishen-gujing can effectively improve the clinical symptoms of CGN patients with spleen-kidney deficiency syndrome and reduce urinary protein.Its clinical disease efficacy and TCM syndrome efficacy are better than those of valsartan group,with good safety.3.Shendi granules can reduce the excretion of podocyte marker proteins PCX and Nephrin in urine of CGN patients with spleen-kidney deficiency syndrome,and reduce the degree of damage to glomerular podocytes.1.Objective Observation of ginseng granules on C-BSA glomerulonephritis model rats foot cell markers PCX,Nephrin and autophagy marker LC3-Ⅱ,Beclin 1 intervention effect,discusses the cords to particles on podocyte damage and the effect of autophagy.2.Methods40 male SD rats were randomly divided into normal group,model group,Shendi granule group and valsartan group,with 10 rats in each group.The cationic bovine serum albumin（C-BSA）nephritis rat model was established except for the normal group.After 28 days of modeling,the urine protein test paper results were（+）++,which confirmed the success of the rat model of C-BSA nephritis.Shendi granules group was given Shendi granules aqueous solution 4.0g/（kg·d）by gavage;Valsartan group was gavaged with Valsartan capsule suspension 20mg/（kg·d）,while normal group and model group were gavaged with normal saline,3ml each time,once a day,for consecutive 8weeks.The urine of rats for 24 hours was collected by metabolic cage,and the blood of abdominal aorta and right kidney were aseptically collected.The concentrations of serum creatinine（SCR）and urea nitrogen（BUN）,urinary albumin and urinary creatinine,and the contents of urinary Nephrin and Podocalyxin were determined by enzyme linked immunosorbent assay（ELISA）.Immune protein printing（Western Blot）in the detection of renal tissue Nephrin,Podocalyxin,Beclin 1,LC3-Ⅱ protein expression level;Immunofluorescence test Beclin 1,Ⅱ LC3-expressed in renal tissue;The changes of glomerular ultrastructure and podocyte autophagosomes were observed by transmission electron microscopy,and the mean thickness of glomerular basement membrane（GBM）（Tg BM）was calculated.3.Results3.1 General situation of rats: rats in the normal group reacted sensitively,had good mental state,bright and dense hair,and had normal eating and drinking.Rats in model group showed poor mental and hair gloss,hair loss,reduced eating and drinking,increased urine volume,sparse stools,and abdominal subcutaneous edema in some rats.The mental state,hair loss,eating and drinking water,stool and other conditions of rats in Shendi granule group and Valsartan group were improved.3.2 Comparison of serum creatinine（SCR）and urea nitrogen（BUN）, urinary albumin/urinary creatinine ratio（ACR）,urinary Nephrin and Podocalyxin（ELISA）: Compared with the normal group,the contents of SCR,BUN,urinary ACR,urinary Nephrin and Podocalyxin in model group were significantly increased（P < 0.05）.Compared with model group,the contents of SCR,BUN,urinary ACR,urinary Nephrin and Podocalyxin in Shendi granule group and valsartan group were significantly decreased（P < 0.05）;The changes of SCR,BUN,urinary ACR and urinary Podocalyxin in Shendi granule group were significantly better than those in valsartan group（P < 0.05）.3.3 Comparison of protein expression of Nephrin,Podocalyxin,Beclin1,LC3-Ⅱ and ratio of LC3-Nat/LC3-Eng in renal tissue of rats（Western blot method）:Compared with the normal group,the expressions of Nephrin and Podocalyxin in renal tissue of model group were significantly decreased,while the expressions of Beclin1,LC3-Ⅱ,LC3-N/LC3-N were significantly increased（P <0.05）.Compared with model group,the expression levels of Nephrin and Podocalyxin in Shendi granule group and valsartan group were significantly increased（P < 0.05）;The expressions of Beclin1,LC3-Ⅱ and LC3-Na/LC3-en in Shendi granularity group were further increased（P < 0.05）,while the expressions of Beclin1,LC3-Con and LC3-Ag/LC3-o in valsartan group were not significantly increased（P > 0.05）.The changes of Podocalyxin,Beclin1,LC3-Ⅱ and LC3-Na/LC3-Na in Shendi granules group were significantly better than those in valsartan group（P < 0.05）.3.4 Beclin1 in renal tissues of rats,LC3-Ⅱ immunofluorescence expression comparison（immunofluorescence）: normal group of autophagy markers in rat kidney Beclin1,LC3-Ⅱ expression is less,and the model group rats Beclin1,LC-Ⅱ expression in kidney tissues significantly enhanced.To particle group compared with model group,and the rat kidney tissues of Beclin1,further enhance the LC3-Ⅱ expression degree and the valsartan group in the rat kidney tissues Beclin1,degree of LC3-Ⅱ expression with the model group rats.3.5 The changes of glomerular ultrastructure and podocyte autophagosomes were observed by transmission electron microscopy, and the mean thickness of glomerular basement membrane（GBM）was calculated:（1）The structure of glomerular podocytes in normal group was clear,no fusion of foot processes was observed,the basement membrane was uniform and no thickens were observed,and no electron density was observed in subepithelium;A few autophagosomes were observed in podocytes.（2）In the model group,most of the foot processes were fused or disappeared,the basement membrane was diffusely thickened,and large electron dense deposits were observed under the epithelium.The number of autophagosomes in podocytes was higher than that in normal group.（3）The degree of glomerular foot process fusion in Shendi granule group and Valsartan group was less than that in the model group,and the basement membrane was slightly thickened,accompanied by a small amount of electron dense deposition.The number of autophagosomes in podocytes in the Shendi granule group was significantly increased compared with that in the model group,while the number of autophagosomes in the valsartan group was approximately the same as that in the model group.（4）Compared with the normal group,the TGBM of model group was significantly increased（P <0.05）.Compared with model group,TGBM in Shendi granule group and Valsartan group were significantly decreased（P < 0.05）.The change of TGBM in Shendi granule group was significantly better than that in valsartan group（P < 0.05）.4.Conclusion1.Podocyte injury is one of the important mechanisms of proteinuria in C-BSA model rats;Activation of autophagy in C-BSA model rats may be a self-protection mechanism after podocyte injury.2.Shendi granules can promote the autophagy activity of podocytes,and reduce the injury of podocytes and pathological changes of kidneys,which may be one of the mechanisms of Shendi granules in reducing proteinuria. |
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