基于多因素造模法改良脾胃气虚型糖尿病胃轻瘫大鼠模型制备方法研究

Study on the Method of Improving the Model Preparation of Diabetes Gastroparesis Rats with Spleen and Stomach Qi Deficiency Based on Multi Factor Modeling

目的:采用多因素造模法联合链脲佐菌素(STZ)配合高糖高脂饲料建立脾胃气虚型糖尿病胃轻瘫(DGP)大鼠模型,并对该模型进行评价。方法:20只SPF级Wistar大鼠依据随机数字表随机分为空白组8只和造模组12只,以多因素造模法复制脾胃气虚型大鼠模型,制定脾胃气虚型模型评价量表,将符合条件大鼠纳入实验;采用小剂量多次注射STZ联合高糖高脂饲料复制DGP大鼠模型,于实验6周末取材。观察大鼠随机血糖、体质量、胃排空率、小肠推进率;Elisa法检测血清中血清淀粉样蛋白(SAA)、D-木糖、乳酸脱氢酶(LDH)的含量;光镜下观察脾脏病理结构改变。结果:与空白组比较,造模组大鼠一般情况(精神状态、粪便、饮食量)较差、体质量降低,差异有统计学意义(P<0.05);造模组随机血糖显著升高,差异有统计学意义(P<0.01);与空白组比较,造模组胃排空率、小肠推进率、SAA、D-木糖、LDH含量降低,差异有统计学意义(P<0.05,P<0.01);光镜下显示造模组脾脏被膜增厚,红髓和白髓分界限不清,淋巴细胞减少。脾胃气虚型DGP证型评价:造模组大鼠有神疲倦怠、体质量降低、蜷缩、活动度差、眯眼、毛色暗淡、舌色偏暗等脾胃气虚证表现。结论:多因素造模法建立的大鼠模型符合脾胃气虚型DGP大鼠标准。

Objective:To establish and evaluate the model of diabetes gastroparesis(DGP)rats with spleen and stomach qi deficiency by multifactor modeling combined with streptozotocin(STZ)and high sugar and high-fat diet.Methods:Twenty SPF grade Wistar rats were randomly divided into a blank group(n=8)and a modeling group of(n=12).The spleen stomach qi deficiency rat model was replicated using a multifactor modeling method,and an evaluation scale for the spleen stomach qi deficiency model was developed.Rats that met the criteria were included in the experiment.The DGP rat model was replicated by injecting small doses of STZ multiple times combined with high sugar and high fat diet,and samples were collected after 6 weeks of experiment commencement.Blood glucose,body mass,gastric emptying rate,and small intestine propulsion rate of rats were randomly observed.Elisa method was used to detect the levels of serum amyloid protein(SAA),D-xylose,and lactate dehydrogenase(LDH)in serum.The pathological structural changes of the spleen were observed under light microscopy.Results:Compared with the rats in blank group,the rats in model group had poorer general conditions(in mental state,defecation,and dietary intake),and decreased body mass,with a statistically significant difference(P<0.05).The random blood glucose levels in the modeling group significantly increased(P<0.01).Compared with the blank group,the gastric emptying rate,small intestine propulsion rate,SAA,D-xylose,and LDH levels in the modeling group decreased significantly(P<0.05,P<0.01).Under light microscopy,it was observed that the splenic capsule of the model group was thickened,the boundary between red and white pulp was unclear,and lymphocytes were reduced.Evaluation of DGP syndrome type of spleen and stomach qi deficiency showed that the model group rats showed symptoms of spleen and stomach qi deficiency,such as fatigue,decreased body mass,curling up,poor mobility,squinting eyes,dull fur color,and dark tongue color.Conclusion:The rat model established by the multi factor modeling method meets the standard of DGP rats with spleen stomach qi deficiency.