理脾复方制剂及硫酸锌对大鼠结肠平滑肌细胞增殖及其脑肠肽的影响

中国中西医结合儿科学 ›› 2021, Vol. 13 ›› Issue (5): 375-380.

理脾复方制剂及硫酸锌对大鼠结肠平滑肌细胞增殖及其脑肠肽的影响

目的
观察10%理脾复方制剂和硫酸锌含药血清对结肠平滑肌细胞（CSMCs）增殖及其脑肠肽的调控作用，探索两者药物治疗厌食的作用机制。
方法：60只幼龄大鼠中取10只培养大鼠CSMCs，余50只大鼠随机分为对照组，低、中、高剂量理脾复方制剂组，硫酸锌组，每组10只。对照组灌胃给予等体积饮用水；硫酸锌组灌胃给予20 mg/kg硫酸锌口服溶液；低、中、高剂量理脾复方制剂组分别灌胃给予2.4、4.8、9.8 g/kg理脾复方制剂，各组每日灌胃1次，连续7 d，末次灌胃2 h后采血制备含药血清。将各组含药血清稀释至10%后作用于大鼠CSMCs 9个时间点，采用八肽胆囊收缩素（CCK8）检测各含药血清组对CSMCs增殖影响；采用酶联免疫吸附法、蛋白质印迹法、实时聚合酶链反应法检测各含药血清组干预大鼠CSMCs最佳增殖起始点时细胞中瘦素（LP）、神经肽Y（NPY）、CCK-8蛋白浓度、蛋白相对表达量、基因相对表达量。
结果:10%不同剂量理脾复方制剂及硫酸锌含药血清均能促进大鼠CSMCs增殖，最佳增殖起始时间为24 h。各含药血清组与对照组比较，各含药血清组均能增加大鼠CSMCs中LP、NPY蛋白浓度、蛋白相对表达量、基因相对表达量（P<005），降低CCK8蛋白浓度、蛋白相对表达量（P<0.05）。
结论:理脾复方制剂和硫酸锌可促进CSMCs增殖，影响CSMC中的LP、NPY、CCK-8表达，可能是其治疗厌食的重要作用机制之一。

550003 贵阳，贵阳市妇幼保健院儿童康复医学科（刘启艳）；551702 贵州毕节，毕节市中医院儿科（詹伟）；554399 贵州铜仁，铜仁职业技术学院附属医院针灸康复科（高娜）；550003 贵阳，贵州中医药大学第二附属医院儿科（张力文，庞平，李春，彭玉）

出版日期:2021-10-25 发布日期:2021-11-15
通讯作者: 彭玉，E-mail：pengyu1022@126.com
作者简介:刘启艳（1985-），女，医学硕士，主治医师。研究方向：中医药防治儿科疾病临床与实验研究
基金资助:
：国家自然科学基金项目（81660808）；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员会科学技术基金项目（gzwjkj2020-1-144）;贵州省“十四五”中医药、民族医药重点建设学科［黔中医药函（2021）95号-6］

Effects of Li-Pi Compound Preparation and zinc sulfate on proliferation of colonic smooth muscle cells of rats and the brain-gut peptide

Objective#br# To observe the effect of 10% LiPi Compound Preparation and zinc sulfate drugcontaining serum on the proliferation of colonic smooth muscle cells(CSMCs) and their regulation effect on brain-gut peptide, and to explore the mechanism of the two drugs in the treatment of anorexia.#br# Methods#br# Among the 60 young rats, the CSMCs of 10 rats were cultured, and the remaining 50 rats were randomly divided into five groups: the control group, the low，medium and high-dose Li-Pi Compound Preparation group and the zinc sulfate group, with 10 rats in each group. The control group was given equal volume of drinking water by gavage. The zinc sulfate group was given 20 mg/kg zinc sulfate oral solution by gavage. The low，medium and high-dose groups were given by gavage 2.4, 4.8 and 9.8 g/kg Li-Pi CompoundPreparations respectively. Each group was given gavage once a day for 7 consecutive days, and blood samples were collected to prepare drug-containing serum at 2h after the last gavage.The drug-containing serum of each group was diluted to 10% and was given to rat CSMCs at 9 time points. The effect of drug-containing serum in each group on the proliferation of colonic smooth muscle cells was detected by CCK-8;Western Bolt, ELISA and RT-PCR were used to detect the relative expression levels of LP,NPY and CCK8 protein concentration in the cells at the optimal starting point of CSMCs proliferation.#br# 〖WT5”HZ〗Results〓#br# The 10% differentdosedrugcontaining serum of LiPi Compound Preparations and zinc sulfate could promote the proliferation of CSMCs in rats, and the optimal starting time of proliferation was 24h.Compared with the control group, each drugcontaining serum group could increase the concentration of LP and NPY protein in rat CSMCs and the relative expression level of protein and gene（P<0.05),reduce the concentration and the relative expressionlevel of CCK8 protein（P<0.05).#br# 〖WT5”HZ〗Conclusion〓#br# LiPi Compound Preparation and zinc sulfate can promote the proliferation of CSMCs,and affect the expression of LP,NPY and CCK8 in CSMCs, which may be one of the important mechanisms in the treatment of anorexia.#br#

Children Rehabilitation Medicine Department, Guiyang Maternal and Child Health Hospital，Guiyang 550003,China

Online:2021-10-25 Published:2021-11-15

摘要/Abstract

摘要： 目的:观察10%理脾复方制剂和硫酸锌含药血清对结肠平滑肌细胞（CSMCs）增殖及其脑肠肽的调控作用，探索两者药物治疗厌食的作用机制。
方法:60只幼龄大鼠中取10只培养大鼠CSMCs，余50只大鼠随机分为对照组，低、中、高剂量理脾复方制剂组，硫酸锌组，每组10只。对照组灌胃给予等体积饮用水；硫酸锌组灌胃给予20 mg/kg硫酸锌口服溶液；低、中、高剂量理脾复方制剂组分别灌胃给予2.4、4.8、9.8 g/kg理脾复方制剂，各组每日灌胃1次，连续7 d，末次灌胃2 h后采血制备含药血清。将各组含药血清稀释至10%后作用于大鼠CSMCs 9个时间点，采用八肽胆囊收缩素（CCK-8）检测各含药血清组对CSMCs增殖影响；采用酶联免疫吸附法、蛋白质印迹法、实时聚合酶链反应法检测各含药血清组干预大鼠CSMCs最佳增殖起始点时细胞中瘦素（LP）、神经肽Y（NPY）、CCK8蛋白浓度、蛋白相对表达量、基因相对表达量。
结果:10%不同剂量理脾复方制剂及硫酸锌含药血清均能促进大鼠CSMCs增殖，最佳增殖起始时间为24 h。各含药血清组与对照组比较，各含药血清组均能增加大鼠CSMCs中LP、NPY蛋白浓度、蛋白相对表达量、基因相对表达量（P<0.05），降低CCK8蛋白浓度、蛋白相对表达量（P<0.05）。
结论：理脾复方制剂和硫酸锌可促进CSMCs增殖，影响CSMC中的LP、NPY、CCK8表达，可能是其治疗厌食的重要作用机制之一。

关键词: 厌食, 理脾复方制剂, 硫酸锌, 结肠平滑肌细胞, 瘦素, 神经肽Y, 八肽胆囊收缩素

Abstract:

Objective：To observe the effect of 10% Li-Pi Compound Preparation and zinc sulfate drug-containing serum on the proliferation of colonic smooth muscle cells(CSMCs) and their regulation effect on braingut peptide, and to explore the mechanism of the two drugs in the treatment of anorexia.

Methods：Among the 60 young rats, the CSMCs of 10 rats were cultured, and the remaining 50 rats were randomly divided into five groups: the control group, the low，medium and high-dose Li-Pi Compound Preparation group and the zinc sulfate group, with 10 rats in each group. The control group was given equal volume of drinking water by gavage. The zinc sulfate group was given 20 mg/kg zinc sulfate oral solution by gavage. The low，medium and high-dose groups were given by gavage 2.4, 4.8 and 9.8 g/kg Li-Pi CompoundPreparations respectively. Each group was given gavage once a day for 7 consecutive days, and blood samples were collected to prepare drug-containing serum at 2h after the last gavage.The drug-containing serum of each group was diluted to 10% and was given to rat CSMCs at 9 time points. The effect of drug-containing serum in each group on the proliferation of colonic smooth muscle cells was detected by CCK-8;Western Bolt, ELISA and RT-PCR were used to detect the relative expression levels of LP,NPY and CCK-8 protein concentration in the cells at the optimal starting point of CSMCs proliferation.

Results：The 10% differentdosedrug-containing serum of Li-Pi Compound Preparations and zinc sulfate could promote the proliferation of CSMCs in rats, and the optimal starting time of proliferation was 24h.Compared with the control group, each drug-containing serum group could increase the concentration of LP and NPY protein in rat CSMCs and the relative expression level of protein and gene（P<0.05),reduce the concentration and the relative expressionlevel of CCK-8 protein（P<0.05).
Conclusion：Li-Pi Compound Preparation and zinc sulfate can promote the proliferation of CSMCs,and affect the expression of LP,NPY and CCK-8 in CSMCs, which may be one of the important mechanisms in the treatment of anorexia.

Key words: Anorexia, LiPi Compound Preparation, Zinc sulfate, Colonic smooth muscle cells, LP, NPY, CCK-8

刘启艳, 詹伟, 高娜, 张力文, 庞平, 李春, 彭玉. 理脾复方制剂及硫酸锌对大鼠结肠平滑肌细胞增殖及其脑肠肽的影响[J]. 中国中西医结合儿科学, 2021, 13(5): 375-380.

LIU Qiyan, ZHAN Wei, GAO Na, ZHANG Liwen, PANG Ping, LI Chun, PENG Yu. Effects of Li-Pi Compound Preparation and zinc sulfate on proliferation of colonic smooth muscle cells of rats and the brain-gut peptide[J]. Chinese Pediatrics of Integrated Traditional and Western Medicine, 2021, 13(5): 375-380.

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