理脾复方制剂对厌食症大鼠下丘脑脑肠肽的影响

Effects of Li-pi compound preparation on brain-gut peptide of hypothalamus in anorexic rats

目的 探讨理脾复方制剂治疗小儿厌食症的作用机制及靶点。方法 50只SPF级SD大鼠,适应性喂养3 d后按照随机数字表法分为空白组(N组)10只和模型组(M组)40只。N组给予常规饲料喂养,M组给予厌食症模型饲料喂养,造模成功后将M组再次按照随机数字表法分成4组,即模型对照组(MC组),理脾复方制剂低、中、高剂量组(LD、D、HD组),每组10只。N组、MC组给予饮用水,LD组、D组和HD组分别给予理脾复方制剂,每日灌胃1次,7 d后禁食24 h,观察每组的摄食量、体质量及下丘脑瘦素(LP)、神经肽Y(NPY)、八肽胆囊收缩素(CCK-8)蛋白浓度、蛋白与mRNA相对表达量。结果 大鼠摄入厌食症模型饲料第5天后摄食量及体质量分别减少48.1%和14.4%,提示造模成功。理脾复方制剂干预后,各组大鼠摄食量均明显增加,HD组>N组>D组>LD组>MC组,其中LD组和D组大鼠摄食量呈逐日增加趋势,HD组摄食量较为稳定;MC组于第3天逐渐增加,但均低于其余各组。各组大鼠每日体质量均明显低于N组,差异均有统计学意义(P<0.01);与MC组比较,LD组和D组体质量增长不明显,差异无统计学意义(P>0.05),HD组于第3天体质量明显增加,差异有统计学意义(P<0.05),第6天增加更显著(P<0.01)。与N组比较,其余各组下丘脑组织中LP、NPY、CCK-8蛋白浓度明显降低,差异均有统计学意义(P<0.01);与MC组比较,LD组、D组及HD组可显著上调下丘脑中LP、NPY、CCK-8蛋白浓度(P<0.01);LD组、D组及HD组两两比较,LP、NPY、CCK-8蛋白浓度差异均有统计学意义(P<0.01),其上调作用与剂量呈正相关,均以高剂量调节作用最为明显。与N组比较,MC组LP蛋白相对表达量增高(P<0.05),NPY与CCK-8蛋白相对表达量下降(P<0.05);与MC组比较,LD组、D组及HD组LP蛋白相对表达量均明显下降,LD组和HD组NPY、CCK-8蛋白相对表达量均明显上升,差异均有统计学意义(P<0.05);LD组、D组及HD组间LP蛋白相对表达量比较,差异均无统计学意义(P>0.05),LD组和HD组的NPY、CCK-8蛋白相对表达量均较D组明显升高(P<0.05),HD组CCK-8蛋白相对表达量较LD组明显升高,差异有统计学意义(P<0.05)。与N组比较,MC组LP mRNA相对表达量明显增高(P<0.05),NPY与CCK-8 mRNA相对表达量明显下降(P<0.05);与MC组比较,LD组、D组及HD组LP mRNA相对表达量均明显下降(P<0.05),NPY mRNA相对表达量明显升高(P<0.05),而仅LD组和D组的CCK-8 mRNA相对表达量明显升高(P<0.05),HD组CCK-8m RNA相对表达量变化不明显(P>0.05)。LD组、D组LP mRNA相对表达量明显低于HD组(P<0.05),但LD组与D组比较,差异无统计学意义(P>0.05);LD组与D组的NPY mRNA及CCK-8 mRNA相对表达量明显高于HD组(P<0.05),且LD组较D组也明显升高(P<0.05)。结论 理脾复方制剂可促进厌食症大鼠摄食量增加、体质量增长,各剂量组均可调控下丘脑中LP、NPY、CCK-8蛋白浓度、蛋白与mRNA相对表达量,这可能是其治疗厌食症的主要作用机制及靶点之一。

Objective To explore the mechanism and targets of Li-pi compound preparation in the treatment of anorexic rats.Methods After 3 days of adaptive feeding,50 SPF SD rats were divided into blank group(N group) with 10 rats and model group(M group) with 40 ones according to random number table method.Group N was fed with conventional feeds and group M was fed with anorexic model feeds;After successfully establishing modeling,M group was divided into 4 groups again according to random number table method,namely model control group(MC group) and low,medium and high dose groups of Li-pi compound preparation(LD,D and HD groups),with 10 rats in each group;N group and MC group were given drinking water,LD group,D group and HD group were given Li-pi compound preparation by gavage once a day,respectively,and fasted for 24 hours after 7 days.The effects of food intake,body mass,hypothalamic leptin(LP),neuropeptide Y(NPY),octapeptide cholecystokinin(CCK-8) protein concentration and relative expression of protein and mRNA were observed.Results After 5 days of ingestion of anorexia model feeds,the food intake and body mass of rats decreased by 48.1% and 14.4%,respectively,indicating that the model was successfully established.After the intervention of Li-pi compound preparation,the food intake of rats in each group increased significantly,HD group>N group>D group>LD group>MC group.The food intake thereof in LD group and D group increased day by day,and the food intake of HD group was relatively stable;That of MC group increased gradually on the 3 rd day,but lower than other groups.The daily body mass of rats in each group was significantly lower than that in N group,and the difference was statistically significant(P<0.01);The increase of body mass in LD group and D group was not significant as compared with MC group,and the difference was not statistically significant(P>0.05).The body mass of HD group increased significantly on the 3 rd day,and the difference was statistically significant(P<0.05),and the increase was more significant on the 6 th day(P<0.01).The protein concentration of LP,NPY and CCK-8 in hypothalamus of other groups decreased significantly as compared with group N,and the difference was statistically significant(P<0.01);LD group,D group and HD group could significantly up-regulate the protein concentration of LP,NPY and CCK-8 in hypothalamus as compared with MC group(P<0.01);Paired comparison among LD group,D group and HD group for protein concentration of LP,NPY and CCK-8 proved significant(P<0.01),whose up-regulation effect was positively correlated with the dose,and the regulation effect of high dose was the most significant.The relative protein expression levels of LP increased(P<0.05) and those of NPY and CCK-8 decreased in MC group as compared with N group(P<0.05);The relative protein expression levels of LP decreased significantly in LD group,D group and HD group as compared with MC group,and the relative protein expression of NPY and CCK-8 increased significantly in LD group and HD group,and the difference was statistically significant(P<0.05);There was no significant difference in the relative protein expression of LP among LD group,D group and HD group(P>0.05).The relative protein expression of NPY and CCK-8 in LD group and HD group increased significantly higher than that in D group(P<0.05),the relative protein expression of CCK-8 in HD group in creased significantly higher than that in LD group,and the difference was statistically significant(P<0.05).The relative expression of LP mRNA significantly increased(P<0.05) whereas that of NPY and CCK-8 mRNA significantly decreased in MC group than those in N group(P<0.05),the relative expression of CCK-8 mRNA only in LD group and D group significantly increased(P<0.05),whereas no significant change was found in HD group(P>0.05).The relative expression of LP mRNA in LD group and D group was significantly lower than that in HD group(P<0.05),but there was no significant difference between LD group and D group(P>0.05);The relative expression of NPY mRNA and CCK-8 mRNA in LD group and D group was significantly higher than that in HD group(P<0.05),and that in LD group was also significantly higher than that in D group(P<0.05).Conclusion Li-pi compound preparation can promote the increase of food intake and body mass of anorexic rats.Each dose group can regulate the protein concentration of LP,NPY and CCK-8 and the relative expression of protein and mRNA in hypothalamus,which may be one of the main mechanisms and targets thereof in the treatment of anorexia.