| ObjectiveThe spleen and stomach damp-heat syndrome, also called "Zhong-jiao damp-heat syndrome", is prevalent in Ling' nan, and generally caused by damp-heat, over-eating fatness and pungent, which may injure the spleen and stomach. It is marked with complicated clinical symptoms, longer course and it's hard to heal completely. So in the occurrence and development of the syndrome, many factors may spur the organism and the body may counteract with the stimulation. The study intends to fabricate the model of spleen and stomach damp-heat syndrome in rats by multiple factors including external damp-heat environment, fat and sweet diet and pathogenic microorganism. We may probe into the change of spleen and stomach damp-heat syndrome in rats, such as thymus index, spleen index, adrenal gland index, plasma CRH and ACTH, serum interleukin-2, interleukin-6 and COR. Meanwhile, the changes of the selected indexes brought by Wang's Lianpuyin are observed. We also explore the impact on neuron-endocrine system of hyperactive hypothalamus-pituitary-adrenal axis. In addition, we will get to know the principle of Qing-re Qu-shi of Wang's Lianpuyin.Method1. Chinese herbal medicine and foodstuffComposition of Wang's Lianpuyin:magnolia bark, golden thread, grassleaf sweetflag rhizome, pinelliae tuber, black curd beans, Gardenia florida, Rhizoma Phragmitis.High glucose and lipid foodstuff:ordinary foodstuff mixed with 12% lard and 8% honey.2. GroupForty male Wister rats were randomly and equally divided into five groups: normal control group, model group, Wang's Lianpuyin group(low, medium and high doses).Each group has 8 rats.3. Creation of modelThis experiment plans to create the model of spleen and stomach damp-heat syndrome in rats by composite factors including external damp-heat environment, fat and sweet diet and pathogenic microorganism.①normal control group:fed with ordinary foodstuff in common environment(20-28℃,50-60% relative humidity);②model group:fed with high glucose and lipid foodstuff, set in simulated climate warehouse and intra-gastric administration with Bacillus coli;③low dosage of Wang's Lianpuyin group:treated with low dosage Wang's Lianpuyin at the base of the model group;④medium dosage of Wang's Lianpuyin group:treated with medium dosage Wang's Lianpuyin at the base of the model group;⑤high dosage of Wang's Lianpuyin group:treated with high dosage Wang's Lianpuyin at the base of the model group.4. Observation and detection of indexesObserve the change of general state, weight, rectal temperature, diet, drinking and shit character between before and after modeling. Compute the index of thymus, spleen and adrenal gland by weighing method and simultaneously detect the concentration of CRH, ACTH in plasma, IL-2, IL-6 and COR in serum of rats by radio-immunity method.5. Statistic methodAll the data were processed by statistical software SPSS 16.0. P<0.05 is setted as the significant level. The data type is quantitative data, expressed with mean士standard deviation(x士s).The data of rectal temperature were analyzed by variance of repeated measure. The indexes of adrenal gland, thymus and spleen, together with other indexes including CRH, ACTH in plasma, IL-2, IL-6 and COR in serum, are analyzed by one-way ANOVA method. Multiple comparison is employed by LSD for different groups.Result1. General condition of the ratsBefore the test, all the rats were in good condition with active movements, glossy white fur, normal eating and drinking, shape stool and normal temperature. After the test, normal group had not abnormal performance. With the conduction of model establishing, most of rats appeared as tired, lazy, dull, fluffy hair, haggard face, reduced diet, sticky stool. From the tenth day, groups employed in composite methods had some abnormal performance, with sticky stool and quickly increased rectal temperature. After entering the simulated climate warehouse, the symptoms were aggravated. As to the groups with different dose of Wang's Lianpuyin, the conditions of movements, diet and stool were improved to normal level.2. The change of temperatureThe data of rectal temperature were analyzed by variance of repeated measure,and the result indicates there is statistical significance in all groups as a whole(F=5.494,P=0.002). In different time points there is statistical significance(F=199.058,P=0.000),there is interaction between grouping and time factor(F=17.935,P=0.000),which means that rectal temperature in different groups appear various tendency with time proceeding. The change of normal control group has no statistic significance at different time points(F=1.088,P=0.381). There is no statistical significance in all groups as a whole on 1st day(P=0.883). The first ten days, the rectal temperature in each group have no significant difference(P=0.493). Rectal temperature of 4 model groups when going into the simulated climate warehouse increased quickly. The temperature of model groups went up obviously after administrated with enter invasive E.Coli. Compared with normal group, it has statistic significance. After giving every dosage Wang's Lianpuyin for 7 days, the rectal temperature of rats decline largely, but model group has no obvious decline.3. Detection of experimental indexesThrough one-way ANOVA, there is statistical significance in plasma CRH of all groups(F=4.157,P=0.000). Through multiple comparison by LSD method,the results indicate as follow. Compared with normal control group, the concentration of plasma CRH in model group elevated, and the variation has statistical significance(P=0.003). Compared with model group, the concentration of plasma CRH in medium dose group declined and the variation has statistical significance(P=0.014). Compared with normal control group, the concentration of plasma CRH in medium dose group heightend, but the variation has no statistical significance(P=0.557). Compared with model group, the concentration of plasma CRH in high and low dose groups declined, but the variation has no statistical significance(P=0.938, P=0.213).Through one-way ANOVA, there is statistical significance in plasma ACTH of all groups(F=5.289,P=0.000). Through multiple comparison by LSD method,the results indicate as follow. Compared with normal control group, the concentration of plasma ACTH elevated, and the variation has statistical significance(P=0.000). Compared with model group, the concentration of plasma ACTH in medium and low dose groups declined and the variation has statistical significance(P=0.001, P=0.020). Compared with normal control group, the concentration of plasma ACTH in medium and low dose groups heightend, but the variation has no statistical significance(P=0.612, P=0.097). Compared with model group, the concentration of plasma ACTH in high dose group declined, but the variation has no statistical significance(P=0.069).Through one-way ANOVA, there is statistical significance in serum Cor of all groups(F=9.563,P=0.000). Through multiple comparison by LSD method,the results indicate as follow. Compared with normal control group, the concentration of serum Cor in model group elevated, and the variation has statistical significance(P=0.000). Compared with model group, the concentration of serum Cor in medium dose group declined and the variation has statistical significance(P=0.001). Compared with normal control group, the concentration of serum Cor in medium dose group heightend, but the variation has no statistical significance(P=0.164). Compared with model group, the concentration of serum Cor in high and low dose groups declined, but the variation has no statistical significance(P=0.627, P=0.120).Through one-way ANOVA, there is statistical significance in serum IL-2 of all groups(F=18.313,P=0.000). Through multiple comparison by LSD method,the results indicate as follow. The concentration of serum IL-2 in model group was lower than in normal control group and the variation has statistical significance (P=0.000).Compared with model group, the concentration of serum IL-2 in medium and low dose groups increased and the variation has statistic significance respectively (P=0.000, P=0.041), and the concentration of serum IL-2 in high dose group increased, but the variation has no statistical significance (P=0.176). The concentration of serum IL-2 in medium dose group was lower than in normal control group and the variation has no statistical significance (P=0.595).Through one-way ANOVA, there is statistical significance in serum IL-6 of all groups(F=15.062,P=0.000). Through multiple comparison by LSD method,the results indicate as follow. The concentration of serum IL-6 in model group was lower than in normal control group and the variation has statistical significance (P=0.000). Compared with model group, the concentration of serum IL-6 in high and medium dose groups increased and the variation has statistic significance respectively (P=0.041, P=0.000), and the concentration of serum IL-6 in low dose group increased, but the variation has no statistical significance (P=0.137). The concentration of serum IL-6 in medium dose group was lower than in normal control group and the variation has no statistical significance (P=0.850).Through one-way ANOVA, there is statistical significance in thymus index of all groups(F=12.837,P=0.000). Through multiple comparison by LSD method,the results indicate as follow. Compared with normal control group, the thymus index in model group declined, and the variation has statistical significance (P=0.000). Compared with model group, the thymus index in medium dose group increased, and the variation has statistical significance (P=0.000), the thymus index in high and low dose groups increased, but the variation has no statistical significance respectively (P=0.587, P=0.273). Compared with normal control group, the thymus index in medium dose group increased, but the variation has no statistical significance (P=0.106).Through one-way ANOVA, there is statistical significance in spleen index of all groups(F=58.847,P=0.000). Through multiple comparison by LSD method,the results indicate as follow. Compared with normal control group, the spleen index in model group declined, and the variation has statistical significance (P=0.000). Compared with model group, the spleen index in medium dose group increased, and the variation has statistical significance (P=0.000), the spleen index in high and low dose groups increased, but the variation has no statistical significance respectively (P=0.174, P=0.616). Compared with normal control group, the spleen index in medium dose group increased, but the variation has no statistical significance (P=0.010).Through one-way ANOVA, there is statistical significance in index of adrenal gland in all groups(F=8.090,P=0.000). Through multiple comparison by LSD method,the results indicate as follow. Compared with normal control group, the index of adrenal gland elevated, and the variation has statistical significance(P=0.000). Compared with model group, the index of adrenal gland medium dose group declined and the variation has statistical significance(P=0.002). Compared with normal control group, the index of adrenal gland in medium dose group heightend, but the variation has no statistical significance(P=0.074). Compared with model group, the index of adrenal gland in high and low dose groups declined, but the variation has no statistical significance(P=0.091, P=0.325).Conclusion1. After the composite action of high glucose and lipid foodstuff, damp-heat environment and pathogenic microorganism, the spleen and stomach damp-heat syndrome animal model appeared to be the same symptoms with the clinical performance of the syndrome.2. Compared with normal control group, the raised concentration of plasma CRH, ACTH, serum Cor in model rats has a significant difference.3. The concentration of serum IL-2 and IL-6 in model group is lower than in normal control group, which suggests that the immunological function, after the composite action of high glucose and lipid foodstuff, damp-heat environment and pathogenic microorganism, is disturbed.4. The variation of thymus index is consistent with that of spleen index, and both of them in model group is lower than in normal control group. The composite action of high glucose and lipid foodstuff, external damp-heat environment and pathogenic microorganism involves the atrophy of thymus and spleen,which means hypo-immunological function.5. The function of adrenal cortex can be reflected by weight of adrenal gland. The index of adrenal gland in model group is bigger than that in normal control group, which indicates the hypertrophy of adrenal gland of spleen and stomach damp-heat syndrome in rats.6. Wang's Lianpuyin may to produce marked effects as follows:1)can recover the general condition of spleen and stomach damp-heat syndrome animal model resulted from composite factors; 2)regulate the function of HPA axis and decrease the secretion of CRH, ACTH and Cor; 3)may increase the concentration of IL-2 and IL-6 in serum;4)may reverse the atrophy of thymus and spleen;5)may curb the hypertrophy of adrenal gland. |
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