The Experimental Study About Theraqpeutie Action And Mechanism Of Huai Qi Huang On Treating IgA Nephropathy Model Rats

PartⅠAn Experimental Study of Treatment of Rat IgA Nephropathy with Huai Qi HuangObjective:To study the therapeutic role of Huai Qi Huang in rat IgA nephropathy.Methods:①The rat IgA nephropathy model was established by use of oral bovine serum albumin(BSA) combined with intravenous injection of Staphylococcal enterotoxin B (SEB) into vein of tail.②After establishment of IgA nephropathy,36 BABL/c mice(male,SPF) were randomised divided into control group, IgAN model group and Huai Qi Huang treatment group. From the 8th week, the rats in Huai Qi Huang treatment group were daily fed with Huai Qi Huang (4g/kg.d) every day. IgAN Model group and control group were daily fed with normal saline.24-hour urine was collected at the 8th,10th and 12th week (before mice execution). to examine the level of urinary protein. Renal function(BUN, Cr), plasma albumin and renal pathology was detected after execution of the mice.Result:①The proteinuria and diffuse mesangial cell proliferation with matrix expansion and IgA deposits were found at the 8th week, indicating successful establishment of IgAN in the rats.②The general performance:IgAN model group showed somewhat loss of appetite, hair messy, body weight loss and etc.. Body weight were no significant different among three groups.③Proteinuria:The urinary protein continued to increase from 8th week to 12th weeks in IgAN model group, The urinary protein level of the model group was higher than the control group and Huai Qi Huang treatment group. The proteinuria decreased after four weeks treatment of Huai Qi Huang, compared with the model group, the difference was statistically significant (P<0.05).④Serum albumin(ALB):Serum albumin decreased obviously in IgAN model group, compared with the control group, the difference was significant (P<0.01). Plasma ALB increased after four weeks treatment of Huai Qi Huang, compared with the model group, the difference was statistically significant (P<0.01).⑤Serum BUN and Cr:The blood urea nitrogen(BUN) and serum creatinine(sCr) were no significant different among the three groups.⑥IgA immunofluorescence examination:There were no IgA deposition in mesangial areas or capillary walls in control group; but strong granular or mass IgA deposits were detected in mesangial regions and capillary walls in model group; The deposits significantly decreased after four weeks treatment of Huai Qi Huang.⑦Microscope examination:The control group showed no abnormal pathological changes in kidney. Diffuse mesangial cell proliferation with matrix expansion and capillary collapse could be seen in model group, but obvious less. after four weeks treatment of Huai Qi Huang. Conlusion:①Oral bovine serum albumin(BSA) combined with intravenous injection of Staphylococcal enterotoxin B (SEB) can cause IgAN in rats.②Huai Qi Huang can attenuate the deposition of IgA, mesangial proliferation and tubulointerstitial lesions, reduce urinary protein, increase serum ALB in IgAN. PartⅡAn Study of Mechanism of Huai Qi Huang in Treating Rat IgA NephropathyObjective:To observe cytokine secretion pattern after Huai Qi Huang treatment of rat IgA nephropathy in order to elucidate its mechanism.Methods:Serum IgA was detected by ELISA. The rat spleen cells were isolated and cultured. The excretion of IL-2, IL-4, INF-y in the supernatant were detected by ELISA. Thl/Th2 balance was represented by calculating the ratio of INF-y/IL-4 in each group.Result:①The level of serum IgA:the average level of serum IgA in model group (449.30±27.27μg/L) was higher than in Huai Qi Huang group (319.52±7.85μg/L) and control group (253.38±13.61μg/L). The difference between model group and control group was statistically significant (P<0.01). Serum IgA was significantly decreased after four weeks treatment of Huai Qi Huang (P<0.05).②Spleen index:the spleen index was higher in Huai Qi Huang group (8.42±1.67) than in control group (8.08±1.47) and model group (8.06±1.52), but the difference were no statistically significant among three groups (P> 0.05).③The level of IL-2 in supernatant of cultured spleen lymphocytes:the average IL-2 level was higher in Huai Qi Huang group (346.05±1.93pg/ml) than in control group (340.32±2.63 pg/ml) and model group (331.81±0.52 pg/ml). The difference between model group and control group was not statistically significant (P> 0.05). The IL-2 levels significantly increased after four weeks treatment of Huai Qi Huang (P<0.01).④The level of IL-4 in supernatant of cultured spleen lymphocytes:the average level of IL-4 was higher in model group (151.82±1.06 pg/ml) than in Huaiqihuang group (149.94±1.78 pg/ml) and control group (117.34±0.51 pg/ ml). The difference between model group and control group was statistically significant (P> 0.05). Althrough IL-2 decreased after four weeks treatment of Huai Qi Huang, but the difference was not statistically significant (P> 0.05).⑤The level of INF-γin supernatant of cultured spleen lymphocytes:the average level of INF-γwas higher in Huai Qi Huang group (1380.65±8.81 pg/ml) than in control group (871.37±8.62 pg/ml) and model group (447.38±20.25 pg /ml). The difference between model group and control group was statistically significant (P<0.01).The level of INF-y significantly increased after four weeks treatment of Huai Qi Huang, even higher than control group (P<0.01).⑥The INF-γ/IL-4 ratio in supernatant of cultured spleen lymphocytes:the average level of INF-γ/IL-4 ratio was higher Huai-Qi Huang group (9.21±0.05) than in control group (7.42±0.05) and the model group (3.14±0.41). The difference between model group and control group was statistically significant (P<0.05). INF-γ/IL-4 ratio significantly increased after four weeks treatment of Huai Qi Huang (P<0.01). Conlusion:①Huang Qi Huai can decrease the level of serum IgA and the deposition of IgA in the kidney.②Huang Qi Huai can attenuate proteinuria and renal lesion in IgAN probably through induce excretion of Th1 type cytokines and restore the balance of Th1/Th2.