理脾复方制剂对大鼠胃窦平滑肌细胞神经肽Y和八肽胆囊收缩素表达的影响  

Effects of Lipi compound preparation on neuropeptide Y and CCK-8 in smooth muscle cells of gastric antrum in rats

作　　者：詹伟 刘启艳 庞平 彭玉 ZHAN Wei;LIU Qiyan;PANG Ping;PENG Yu(Department of Pediatrics,Bijie Hospital of Traditional Chinese Medicine,Bijie,Guizhou,551700,China;Department of Child Healthcare,Guiyang Maternal and Child Health Care Hospital,Guiyang,Guizhou,550003,China;Department of Pediatrics,The Second Affiliated Hospital of Guizhou University of Chinese Medicine,Guiyang,Guizhou,550003,China)

机构地区：[1]毕节市中医院儿科,贵州毕节551700 [2]贵阳市妇幼保健院儿童保健科,贵州贵阳550003 [3]贵州中医药大学第二附属医院儿科,贵州贵阳550003

出　　处：《中医儿科杂志》2021年第2期1-6,共6页Journal of Pediatrics of Traditional Chinese Medicine

基　　金：国家自然科学基金地区基金项目(8166150808);毕节市科技局联合基金项目(毕科联合字Z[2019]2号);2020年贵州省卫生健康委科学技术基金项目(gzwjkj2020-1-144)。

摘　　要：目的观察理脾复方制剂治疗小儿厌食症的可能作用机制。方法60只SD大鼠随机分为生理盐水组,理脾复方制剂低、中、高剂量组,硫酸锌组与细胞培养组共6组,每组10只。细胞培养组主要用于培养原代大鼠胃窦平滑肌细胞,其余各组分别灌胃给药理脾复方制剂、硫酸锌7 d,每日1次,于末次灌胃1 h后进行麻醉下股动脉采血,制备含药血清。将各组含药血清干预作用于大鼠胃窦平滑肌细胞,用细胞增殖实验检测最佳细胞增殖时间,于最佳细胞增殖时间点处,运用实时荧光定量聚合酶链式反应(real-time PCR)法和蛋白免疫印迹法(Western bolt)检测大鼠胃窦平滑肌细胞神经肽Y(NPY)、八肽胆囊收缩素(CCK-8)的基因和蛋白表达。结果含药血清作用后,大鼠胃窦平滑肌细胞最佳增殖时间为12 h。与生理盐水组比较,其余各组NPY mRNA和蛋白相对含量水平均有不同程度升高(P<0.05),CCK-8 mRNA和蛋白相对含量水平均有不同程度降低(P<0.05),其中理脾复方制剂高剂量组和硫酸锌组疗效最佳(P<0.05)。结论理脾复方制剂可能通过上调NPY和下调CCK-8的表达对大鼠胃窦平滑肌细胞分泌的脑肠肽进行调控,进而增强胃窦平滑肌收缩,达到促进胃排空作用,这可能是其治疗厌食症的作用机制。Objective To observe the possible mechanism of Lipi compound preparation in the treatment of children with anorexia.Methods Sixty SD rats were randomly divided into 6 groups,including normal saline group,low,medium and high dose Lipi compound preparation group,zinc sulfate group and cell culture group,with 10 rats in each group.The cell culture group was mainly used to culture primary rat gastric antrum smooth muscle cells.The rest groups were given Lipi compound preparation and zinc sulfate by gavage for 7 days,once a day,1 hour after the last gavage,blood was collected from femoral artery under anesthesia,and then the medicated serum was prepared.The intervention effects of medicated serum of each group was conducted on the smooth muscle cells of gastric antrum in rats.Cell counting kit 8 was used to detect the optimal time of cell proliferation.At the optimal time point of cell proliferation,the gene and protein expression of neuropeptide Y(NPY)and cholecystokinin octapeptide(CCK-8)were detected by real time fluorescent quantitative polymerase chain reaction(real-time PCR)and Western blot.Results After the effects by medicated serum,the optimal proliferation time of rat gastric antrum smooth muscle cells was 12 h.Compared with the normal saline group,NPY mRNA and relative protein levels in the other groups increased in varying degrees(P<0.05),CCK-8 mRNA and relative protein levels decreased in varying degrees(P<0.05),and the high-dose Lipi compound preparation and zinc sulfate group therein had the best effect(P<0.05).Conclusion Lipi compound preparation can regulate the brain gut peptide secreted by rat gastric antrum smooth muscle cells by up regulating the expression of NPY and down regulating the expression of CCK-8,and then enhances the contraction of gastric antrum smooth muscle and promotes gastric emptying,which may be the action mechanism of its therapeutic effect on anorexia.

关 键 词：厌食症 大鼠 理脾复方制剂 胃窦平滑肌细胞 神经肽Y 八肽胆囊收缩素 含药血清 实验研究 

分 类 号：R965.1[医药卫生—药理学]