Clinical Effect Of Modified Huangqi Chifeng Decoction On IgA Nephropathy Proteinuria And Its Protective Effect On Podocytes

Part One:Clinical Reseach:Effect of Modified Huangqi Chifeng Decoction on Prote:inuria and urinary podocyte related proteins in IgA Nephropathy PatientsObjective:To observe the clinical efficacy of Modified Huangqi Chifeng Decoction(MHCD)in the treatment of proteinuria in IgA nephropathy patients and its protective effect on urinary podocyte related proteins by means of a randomized controlled trial.Methods:Sixty patients with IgA nephropathy(stage CKD1-2)with mild to moderate proteinuria and Traditional Chinese Medicine(TCM)syndrome differentiation as qi deficiency and blood stasis combined with wind pathogen and heat toxin were randomly divided into the experimental group and the control group with 30 cases each.The control group was given telmisartan and basic treatment,the experimental group in addition to telmisartan and basic treatment,plus MHCD.The score of TCM symptom and the 24-hour urine protein of the patients were recorded every 4 weeks,and the experimental period was 16 weeks.Blood routine and liver and kidney function were detected at week 0,8 and 16,and podocyte related proteins Nephrin and Podocalyxin(PODXL)were detected at week 0 and 16.Results:?Comparison of 24hU-TP between the experimental group and the control group:After 8 weeks of taking the drug,the 24-hour urinary protein quantification in the treatment group and the control group decreased,respectively,compared with that before treatment(P<0.05).After 16 weeks of taking the drug,the treatment group and the control group had a significant decrease in 24-hour urinary protein quantification compared with that before treatment in each group(P<0.01).After treatment for 8 weeks and 16 weeks,the treatment group was compared with the control group,and the difference was statistically significant(P<0.01).?Comparison of TCM symptom scores between the experimental group and the control group:The symptom scores of the treatment group were significantly lower than those before treatment after 8 weeks and 16 weeks of treatment,respectively,and the difference was statistically significant(P<0.01);the control group was dosed at 8 and 16 weeks.Compared with before treatment,the syndrome scores did not decrease significantly afterwards,and the difference was not statistically significant(P>0.05).After 8 weeks and 16 weeks,the scores of TCM symptoms in the treatment group decreased significantly compared with the control group(P<0.01).?Comparison of HGB,Scr,ALT and AST between the experimental group and the control group:The difference between the two groups after 8 weeks and 16 weeks of HGB treatment was not statistically significant compared with that before treatment(P>0.05).Scr in the experimental group decreased after 8 weeks and 16 weeks respectively compared with before treatment,and the difference was statistically significant(P<0.01);No statistically significant difference was found in the control group(P>0.05).There was no statistically significant difference between the two groups of ALT and AST after 8 weeks and 16 weeks of treatment compared with before treatment in this group(P>0.05).?Comparison of the efficacy of TCM syndromes between the experimental group and the control group::The efficacy of TCM syndromes was compared between the treatment group and the control group after 16 weeks of treatment.The total effective rate of the treatment group was 89.66%,and the total effective rate of the control group was 13.79%.The difference between the two groups is statistically significant(P<0.01).?Comparison of clinical efficacy between the experimental group and the control group::The clinical efficacy of the treatment group was compared with that of the control group after 16 weeks of treatment.The total effective rate of the treatment group was 93.1%and the control group was 65.52%.The difference between the two groups is statistically significant(P<0.05)?Effect of urinary podocyte related protein between the experimental group and the control group::In the treatment group,Nephrin expression was significant(P<0.01)and PODXL expression was statistically significant(P<0.01).After treatment,Nephrin expression was significant(P<0.01)and PODXL expression was statistically significant(P<0.01).Conclusion:MHCD can improve the TCM symptoms of IgA nephropathy patients,reduce the 24-hour urine protein,and reduce the excretion of Nephrin and PODXL.During the observation period,there were no abnormal hemoglobin and adverse reactions of liver and kidney function damage,which indicated that MHCD was effective and safe in the treatment of proteinuria of IgA nephropathy,it could reduce the excretion of urinary podocytosis related proteins Nephrin and PODXL,which had a protective effect on podocytes.Part Two:Experimental Reseach:Protective effect of Modified Huangqi Chifeng Decoction on podocytes in IgA nephropathy ratsObjective:To observe the effect of MHCD on the expressions of podocyte related proteins Nephrin and Podocalyxin in rats with IgA nephropathy.Methods:Bovine serum albumin(BSA)+carbon tetrachloride(CCL4)+lipopolysaccharide(LPS)were used to make the IgA nephropathy rat model.After successful modeling,60 rats were randomly divided into blank group,model group,telmisartan group,MHCD high dose group,MHCD medium dose group,MHCD low dose group,(the three groups are referred to as Chinese medicine high,medium and low dose groups)according to the random number table method,each group has 10 rats.Each group of rats was given corresponding drug intervention,and the administration period was 8 weeks.The experiment lasted for 18 weeks.The 24-hour urinary protein ration was measured every 2 weeks.At the end of the experiment,the abdominal aorta was used to collect blood and remove the rat kidney.After centrifugation,the blood samples were collected for serum,and the levels of Scr,BUN,ALB,ALT and AST were detected.Kidney tissue was rapidly removed after blood collection from the abdominal aorta.After the collection of renal tissue,the pathological changes of glomerulus in each group were observed by special staining under the light microscope(including HE staining,PAS staining and Masson staining),and the degree of renal pathological damage was determined.IgA fluorescence deposition in glomerular mesangial region of each group was observed under immunofluorescence and the intensity of IgA deposition was evaluated by semi-quantitative criteria.Western-Blot method was used to detect the podocyte related proteins Nephrin and Podocalyxin(PODXL)in the kidneys of each group of rats.Results:?24h-UTP in rats at modeling stage:From the 6th week of the experiment,compared with the control group at the same time,24hU-TP in the modeling group increased,and the difference was statistically significant(P<0.05).After the drug intervention,compared with the model group at the same period,the 24hU-TP in telmisartan group and the high dose,medium dose and low dose groups of traditional Chinese medicine all decreased,showing a statistical difference(P<0.05)Compared with telmisartan group at the same period,the 24-hour urinary protein quantification was reduced in the high,medium and low dose groups with statistical difference(P<0.05).Compared with the high dose group,there was no statistical difference in 24hU-TP between the middle dose group and the low dose group(P>0.05)?Blood biochemical indicators of rats in each group:There was no significant difference in Scr,BUN,ALB,ALT,and AST among the groups(P>0.05).?Kidney structure under light microscope of rats in each group:Blank group:normal glomerular structure,clear vascular bulb,good filling,normal distribution of mesangial matrix and mesangial cells,no proliferation,no balloon cavity dilation or atrophy;renal tubules are lined with cubic epithelium,no degeneration and necrosis,regular lumen arrangement,no expansion,no interstitial fibrous tissue hyperplasia;Model group:glomeruli exhibit compensatory expansion,manifested as vascular bulbs swelling,lobulated or nodular lesions,narrowed balloon cavity,local glomerular atrophic lesions,manifested by smaller vascular bulbs,occlusion of the lumen,proliferation of mesangial matrix and mesangial cells,and thickening of the basement membrane;Vesicular-like degeneration of local tubular epithelial cells,disordered tubular arrangement,compensatory dilatation of local tubules,interstitial congestion and edema,and fibrous tissue hyperplasia.Telmisartan group:The glomerulus is slightly compensatory dilated,the degree of angiosphere swelling is reduced,the balloon cavity is slightly narrowed,a small number of glomeruli are atrophic,mesangial matrix and mesangial cells proliferate,and basement membrane is slightly thickened;Focal tubular epithelial cells are vacuolated,the arrangement of renal tubules is more orderly,some tubules are compensatingly dilated,interstitial congestion and edema,a small amount of fibrous tissue hyperplasia.Chinese medicine high dose group:the glomerulus dilated slightly,the degree of swollen blood vessels was significantly reduced,the balloon cavity was slightly narrowed,individual glomeruli were atrophic,mesangial matrix and mesangial cells proliferated,and the basement membrane was slightly thickened;A small amount of tubular epithelial cells with vacuole-like degeneration,neat tubular arrangement,compensatory dilatation of tubules is not obvious,interstitial mild congestion and edema,and a small amount of fibrous tissue hyperplasia.Chinese medicine medium and low dose groups:glomerular compensatory dilatation,the degree of swollen glomeruli is reduced,the balloon cavity is narrowed,a small number of glomeruli are atrophic,mesangial matrix and mesangial cells are slightly proliferated,basal membrane thickening;focal tubular epithelial cell vacuole-like degeneration,slightly irregular tubular arrangement,individual tubular compensation compensatory expansion,interstitial congestion and edema,and a small amount of fibrous tissue hyperplasia.?IgA deposition under fluorescence microscope of rats in each group:The fluorescence intensity of the blank group rats was-?+;the fluorescence intensity of the model group rats was++?+++;the fluorescence intensity of the telmisartan group was+?+++;the fluorescence intensity of the high and medium dose groups of Chinese medicine was +?++;and the fluorescence intensity of the low dose group of Chinese medicine was+?+++,IgA fluorescence deposition intensity was measured by non-parameters of multiple grades of data,the IgA fluorescence deposition intensity of each group was not equal,and the difference was statistically significant.Pairwise comparison:Compared with the blank group,the IgA fluorescence deposition(P<0.05)of the model group was statistically different(P<0.01);compared with the model group,the IgA fluorescence deposition intensity of the telmisartan group and the high and medium dose groups of Chinese medicine,there were statistical differences(P<0.05),and the low dose Chinese medicine group did not show statistical differences(P>0.05);Compared with the telmisartan group,there was no statistical difference in the intensity of IgA fluorescence deposition in the high,medium and low dose groups of Chinese medicine(P>0.05);Compared with the high dose group of Chinese medicine,there was no statistical difference in the IgA deposition intensity of the middle and low dose groups of Chinese medicine(P>0.05);compared with the Chinese medicine medium dose group,the Chinese medicine low dose group showed no significant difference in IgA deposition intensity(P>0.05).?Podocyte related protein expression:Compared with the blank group,the expression of Nephrin and PODXL in the model group decreased significantly(P<0.01);Compared with the model group,the expression in the high dose group increased significantly,and the difference was statistically significant(P<0.01).Compared with the model group,the expression of telmisartan group and the low dose and medium dose groups increased,with no statistical difference(P>0.05).Compared with the low dose group,the expression in the high dose group increased significantly,and the difference was statistically significant(P<0.01).Compared with the medium dose group,the expression in the high dose group all increased,and the difference was statistically significant(P<0.05).Compared with telmisartan group,the expression of PODXL in the high dose group increased,with significant statistical difference(P<0.01),and the expression of Nephrin increased,with statistical difference(P<0.05).Conclusion:MHCD can reduce proteinuria in IgA nephropathy model rats,reduce renal pathological injury,up-regulate the expression of renal podocyte-related proteins Nephrin and PODXL,and have a protective effect on glomerular podocytes.