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ABSTRACT OBJECTIVE：To explore the dose-effect relationship and mechanism of protective effects of total asiaticoside（TA）
on gastrointestinal motility and enteric nervous system（ENS）in aged functional dyspepsia（FD）model rats. METHODS：Aged
male SD rats of 16 months old were randomly divided into blank control group，model group，TA low dose，medium dose and high
dose groups（15，30，60 mg/kg），with 8 rats in each group. FD model was established by tail-stimulation combined with irregular
diet for 4 weeks. The next day after modeling，administration groups were given relevant doses of TA solution intragastrically；
control group and model group were given constant volume of normal saline intragastrically，once a day，for consecutive 15 d.
Gastric emptying rate and small intestinal propulsion rate of rats were examined. ELISA were used to detect serum contents of MTL
and VIP. Immunofluorescence and immunohistochemistry were proposed to measure the expression of ENS marker（S100 β and
GDNF）in gastric antrum tissue. The protein expression of S100 β ，GFAP，PGP9.5，GDNF，p-MEK and p-ERK1/2 in gastric
antrum tissue were measured by Western blotting assay. RESULTS：Compared with blank control group，gastric emptying rate and
small intestinal propulsion rate，serum MTL content and protein expression of PGP9.5 in gastric antrum tissue of model and TA
low，medium dose group were decreased significantly，while serum VIP content，protein expressions of S100β，GFAP，GDNF，
p-MEK and p-ERK1/2 in gastric tissue were increased significantly（P＜0.05）. Compared with model group，gastric emptying rate
and small intestinal propulsion rate of TA groups were increased significantly（P＜0.05）；except for GFAP protein in TA low dose
group（P＞0.05），the serum MTL content and the expression of PGP9.5 protein in gastric antrum tissue of rats in TA groups were
increased significantly，while serum VIP content，protein expression of S100β，GFAP，GDNF，p-MEK and p-ERK1/2 in gastric
antrum tissue were decreased significantly（P＜0.05）. Some or most of the content of gastrointestinal motility indexes and related
factor protein expression were significantly different among TA groups（P＜0.05），and the indexes in TA high dose group could
recover to the levels which were not significantly different with blank control group （P＞0.05）. CONCLUSIONS：TA can
dose-dependently improve the gastrointestinal motility deficiency and ENS dysfunction in aged FD model rats，especially in high
dose （60 mg/kg）of TA group. Its mechanism may be related with promoting the release of endogenous MTL，inhibiting the
secretion of VIP，expression of GDNF and the activation of downstream signaling pathway，and promoting the repair of ENS and
intestinal neurons.
KEYWORDS Total asiaticoside；Functional dyspepsia；Gastrointestinal motility；Enteric nervous system；GDNF；Mechanism；
Dose-effect relationship；Aged rat

功能性消化不良（Functional dyspepsia，FD）是一组 胞外信号调节激酶（ERK）/丝裂原活化蛋白激酶激酶
源自上腹部，持续或反复存在腹痛、腹胀、烧灼感或早饱 （MEK）信号通路相关因子的变化，探究积雪草总苷对老
感的症候群 。流行病学资料显示，我国FD发病率高达 年 FD 模型大鼠保护作用的量效关系和可能机制，为临
[1]
18％～35％ ，其中60岁以上的老年人由于胃肠动力障 床治疗老年FD疗提供实验基础和理论依据。
[2]
碍、胃酸分泌紊乱等生理功能退化，成为了FD的高危人 1 材料
群 。临床上除了使用抗酸药和促动力药治疗FD外，中 1.1 仪器
[3]
医药及针灸等补充治疗方法也逐渐受到越来越多学者 BT25S 型电子天平（德国 Sartorius 公司）；Multiskan
的关注。积雪草总苷是包含积雪草苷、羟基积雪草苷、 MK3 型酶标仪、Sorvall ST 8 型台式离心机（美国 Ther-
积雪草苷B等在内的三萜类皂苷成分混合提取物，近年 mo Fisher Scientific 公司）；Cryo3DM 型冷冻切片机（日
来大量研究表明，积雪草总苷在治疗胃溃疡和胃癌等胃 本Sakura公司）；AiryScan LSM800型激光共聚焦显微镜
肠道疾病方面具有一定作用 [4-6] ，但其对老年 FD 的治疗 （德国Zeiss公司）；IX53型倒置显微镜（日本Olympus公
作用尚未见研究报道。以往的研究表明，胃肠活动的调 司）；Tanon-1200 型凝胶成像系统、EPS600 型电泳仪（上
控和支配同时受到肠神经系统（ENS）、中枢神经系统 海天能科技有限公司）；UPH-I-5/10/20T型台式标准型超
（CNS）、胃肠起搏细胞和平滑肌等在内的神经冲动传导 纯水机（西安优普仪器设备有限公司）。
网络的控制，其中ENS作为由肠神经元和神经胶质细胞 1.2 药品与试剂
（EGCs）构成的独立单位，可不依赖 CNS 发挥对胃肠道 积雪草总苷原料药（广西昌洲天然药业有限公司，
[7]
活动的独立调控作用 。另有研究证实，胶质细胞源性 批号：84696219，含量：95％）；胃动素（MTL）酶联免疫吸
神经营养因子（GDNF）的异常表达也与FD的发生、发展 附测定（ELISA）检测试剂盒（批号：20180318）、血管活
[8]
有关 。基于此，本研究采用积雪草总苷干预老年FD模 性肠肽（VIP）ELISA 检测试剂盒（批号：20180430）均购
型大鼠，并通过检测大鼠体内ENS特异性标志物中枢神 自南京建成生物工程研究所；GFAP兔单克隆抗体、S100
经 特 异 性 蛋 白 S100 β 、胶 质 细 胞 原 纤 维 酸 性 蛋 白 β兔单克隆抗体、PGP9.5兔单克隆抗体、GDNF兔单克隆
（GFAP）、蛋白基因产物 9.5（PGP9.5），以探究积雪草总 抗体、磷酸化 MEK（p-MEK）兔单克隆抗体、磷酸化
甘对 ENS 的保护作用；同时，通过考察 GDNF 及相关细 ERK1/2（p-ERK1/2）兔单克隆抗体（批号分别为GB11096、

·1430 · China Pharmacy 2020 Vol. 31 No. 12 中国药房 2020年第31卷第12期