| Objectives:To explore an animal model of nephritis hematuria that more compatible to the Damp-Heat syndrome and less expensive in cost and more sticking to the "Syndrome closely related to disease" via comparing indexes in different modeled rats.Methods:50 SPF male Wistar rats were used to establish models of nephritis hematuria with damp-heat syndrome, rats were give different treatment based on intragastric administration and regular tail vein injection of bovine serum albumin (BSA), they ware randomly divided into five groups as follows:①normal group;②model group one, injected staphylococcal enterotoxin B (SEB) and fed with high-carbohydrate diet, high fat diet, under an environment of high temperature and high humidity;③model group two, an improved model group which was injected SEB and given damp-heat factors of environment and diet;④model group three, given damp-heat factors of environment and diet and no SEB;⑤model group four: injected SEB and given twice damp-heat factors of environment and diet. Animals were raised under observation and killed at the end of 12 weeks. For each group, general condition, body temperature and the body weight, urine red blood cells, 24-hour urine protein, kidney pathological changes of rats were observed, along with the renal function, 4 items of blood lipids (total serum cholesterol, serum triglycerides, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol), and serum IL-6 content in rats were detected by biological methods, RIA, optical microscopes.Results:1. General status:It appears that rats in each groups manifested fever temperature, preferring lying, sluggish action, erecting hair, drinking less, loss of appetite, congestive inflamed anus, rarefaction stool, urine dark in color and less in volume, dark red tongue, some slightly white and greasy fur. Some of the rats in the group two and four have gum in the eyes, diarrhea, skin edema performance, and the phenomenon of subcutaneous edema, urine in dark color and less volume continued to the end of the experiment. Body weight of rats in group three and four were increased significantly than the other groups to the end of the experiment. The rats net value-added of body temperature in model group that given damp-heat factor was higher than that in other control model group (group one) and the differences were significant (P <0.05).2. Detection of urine red blood cell and urine protein:2.1 Urine red blood cells: From the end of the 5th and the end of 12th weeks, as compared with normal group, the number of urine red blood cells increased significantly in each model groups, the differences were significant (P <0.01), and continued to the end of the experiment. At the end of the 12th week, as compared with the other groups, the group two had the largest number of urine red blood cells, and had a significant difference from other model groups. Urine red blood cells of group three and four were less than group one, and also had a significant difference (P <0.01, P <0.05), moreover, the group four had more than the group three, The difference was not significant (P> 0.05).2.2 Urine protein: At the end of the 12th week, as compared with the other model groups, the model group urine protein was significantly higher than normal group, it had a significant difference (P<0.01). Group two had the largest number of urine protein among all group. The difference between group and group one was not significant (P>0.05). The differences between group one and group three and four were significant (P>0.05), but the difference between model group four and group three was not significant (P>0.05) though model group four demonstrated seemingly higher.3. Determination of renal function3.1 Blood urea nitrogen level (BUN): The level of BUN in model groups were higher than the normal group, the differences were significant (P< 0.05, P<0.01). The level of BUN in group one was highest, but as compared to other groups, it had no significance (P>0.05). The level of group four was higher than group three, but with no significance (P>0.05).3.2 Level of blood Scr: The model groups were higher than the normal group obviously, and the differences were significant (P<0.01). The level of group two was highest, and as compared to group one, the difference was significant (P<0.01), the level of group three and four were higher than group one, but the difference was not significant(P>0.05).The level of group four was higher than group three, but the difference wasn't significant(P>0.05).4. Determination of blood-lipid level 4.1 Serum total cholesterol (CHOL): the level of CHOL model groups was higher than normal groups, the differences was significant (P<0.01). The level of group two was highest, and the difference was significant compared to group one (P< 0.01), the level of group three and four were higher than group one, but the difference was not significant (P>0.05). The level of group four was higher than group three, but the difference was not significant (P>0.05).4.2 Serum total triglyceride(TG): the level of TG model groups was higher than normal groups, the differences was significant(P<0.01).The level of group two was highest, and the difference was significant as compared to group one(P<0.01),the level of group three and four were higher than group one, but the difference wasn't significant(P>0.05). The level of group four was higher than group three, but the difference wasn't significant (P>0.05).4.3 Serum total high-density lipoprotein (HDL-C), serum total low-density lipoprotein (HDL-C): the difference of the level in HDL-C and HDL-C among all the groups weren't significant (P>0.05).5 The level of IL-6:The level of IL-6 in all model groups was higher than normal groups, and the difference was significant (P< 0.01) ; The level of group two was highest, and the difference was significant compared to group one(P< 0.01),the level of group three and four were higher than group one, but the difference wasn't significant(P>0.05).The level of group four was higher than group three, but the difference wasn't significant(P>0.05).6. Observation on Histopathology:Histological changes were observed under light microscope, we can see the mesangial cells and glomerular basement membrane were not proliferate, the capillary loop open. the structure of glomerular atrophy tubular is normal, and stromal hyperplasia and infiltration of inflammatory cells were not observed in the normal group. In the other model groups, diffuse mesangial area was seen widened. The mesangial cells and glomerular basement membrane were proliferating, cavity of capillaries became narrow , Some of the glomerulars atrophied, some was phyllodes, renal capsule mildly expanded, tubulointerstitial have infiltration of inflammatory cells, especially the pathological changes of group two were the most severe, no significant difference was seen between model group one and model group two. The model group three and model group four showed similar changes to that of model group one and two, but with less significance. No obvious difference was observed in comparison between model group three and four.Conclusions:According to the damp-heat pathogen of the Chinese medicine theory, using the composite factor, we successfully established rat models of nephritis hematuria with damp-heat syndrome. When compared with traditional method, exerting the factor of diet and damp-heat environment can raise the temperature of rat, make a more severe hematuria edema, higher contents of Scr, triglyceride, cholesterol, IL-6, creatinine, and made the damp-heat symptom more prominently. Whether from the manifestation of symptom to the characteristic of disease or from the onset condition to microcosmic norm, this model, which is similar to human damp-heat nephritis, is a better animal hematuria one for damp-heat nephritis with the "syndrome closely related to disease". Animal model with damp-heat syndrome of nephritis hematuria, which was given no SEB injection and only more food and damp-heat factor, was an animal model whose method was cheaper and more typically than others. |
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