中药理脾复方对厌食大鼠结肠作用机制研究

Effect and mechanism of Li-Pi Compound Preparation on colon in anorexic rats

目的探讨中药理脾复方对厌食大鼠结肠的神经肽Y(NPY)、胆囊收缩素8肽(CCK8)和瘦素(LP)表达影响。方法以病因模拟法建立大鼠厌食模型。按照体重将造模成功大鼠随机分为5组:模型组、对照组和低、中、高3个剂量实验组,每组10只;另取10只正常大鼠作为正常组。正常组与模型组均灌胃给予等体积饮用水;对照组灌胃给予硫酸锌口服溶液20 mg·kg^-1;低、中、高3个剂量实验组分别灌胃给予中药理脾复方2.4,4.8和9.6 g·kg^-1,每天给药1次,连续1周。用酶联免疫吸附法检测结肠组织NPY蛋白浓度,用蛋白质印迹法检测结肠组织CCK8蛋白相对表达水平(灰度值),实时定量-PCR法检测结肠组织LP基因的表达水平(△Ct值)。结果正常组、模型组、对照组和低、中、高3个剂量实验组的NPY蛋白浓度分别为(251.65±10.83),(72.42±22.58),(212.01±20.67),(87.00±4.33),(137.59±8.31)和(206.28±14.95)pg·mL^-1;上述6组的CCK8蛋白相对表达量分别为0.83±0.04,0.84±0.02,0.93±0.01,0.99±0.02,1.00±0.02和0.92±0.02;上述6组的LP基因表达水平分别为1.02±0.20,1.24±0.02,0.96±0.17,1.33±0.07,1.01±0.24和1.07±0.08。上述指标:模型组与正常组相比,差异均有统计学意义(均P<0.05);对照组和低、中、高3个剂量实验组与模型组相比,差异均有统计学意义(均P<0.05)。结论中药理脾复方可显著增加结肠组织中CCK8、NPY和LP水平表达,调控CCK8、NPY和LP蛋白及基因水平的表达,这可能是其治疗小儿厌食症的作用机制之一。

Objective To study the effects of Li-Pi Compound Preparation on the expression of colon of cholecystokinin 8 peptide(CCK8),leptin(LP) and neuropeptide Y(NPY)in anorexia rats. Methods The model of anorexia in rats was established by etiological simulation,and rats after modeling were randomly divided into five groups:model group,control group and experimental-L,-M,-H doses experimental groups with 10 cases per group. In addition, another 10 healthy rats were selected as normal group. The normol group and model group were given drinking water by gavage. The control group was given 20 mg·kg^-1 of oral solution of zinc sulfate by gavage. At the dose of 2.4,4.8 and 9.6 g·kg^-1 Li-Pi Compound Preparation were given by gavage respectively in the experimental-L,-M,-H groups. Rats in the six groups were administrated once a day for one week. The concentration of NPY protein in colon tissue was detected by enzymelinked immunosorbent assay. The relative expression( gray value) of CCK8 protein was detected by Western blot. The relative expression( △Ct value) of LP gene was detected by real time quantitative-PCR. Results The expression concentration of NPY in normal group,model group,control group and experimental-L,-M,-H groups were( 251. 65 ± 10. 83),( 72. 42 ± 22. 58),( 212. 01 ± 20. 67),( 87. 00 ± 4. 33),( 137. 59 ± 8. 31) and( 206. 28 ± 14. 95)pg·mL^-1;the relative expression of CCK8 protein in the six groups were 0. 83 ± 0. 04,0. 84 ± 0. 02,0. 93 ± 0. 01,0. 99 ± 0. 02,1. 00 ± 0. 02 and 0. 92 ± 0. 02;the relative expressionof LP gene in the six groups were 1. 02 ± 0. 20,1. 24 ± 0. 02,0. 96 ± 0. 17,1. 33 ± 0. 07,1. 01 ± 0. 24 and 1. 07 ± 0. 08. Compared between model group and normal group,the difference of the factors were statistically significant( all P < 0. 05);compared between control group,experimental-L,-M,-H groups and model group,the difference of the factors were statistically significant( all P < 0. 05). Conclusion Li-Pi Compound Preparation can significantly increase the expression of CCK8,NPY and LP levels in colon tissue and regulate the expression level of CCK8,NPY and LP protein and gene,which may be one of the mechanisms of its treatment of infantile anorexia.