南京中医药大学附属苏州市中医医院,江苏省苏州市姑苏区沧浪新城杨素路18号,215009
wxpeng2004@163.com
收稿:2025-10-23,
修回:2026-03-05,
录用:2025-12-18,
纸质出版:2026-05-02
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王楚越,李家诚,杨应奇等.通便汤对慢传输型便秘模型大鼠结肠组织VAPB-PTPIP51复合物及Cajal间质细胞自噬的影响[J].中医杂志,2026,67(09):985-993.
WANG Chuyue,LI Jiacheng,YANG Yingqi,et al.Effect of Tongbian Decoction (通便汤) on the VAPB-PTPIP51 Complex and Autophagy of Interstitial Cells of Cajal in the Colon of Slow Transit Constipation Model Rats[J].Journal of Traditional Chinese Medicine,2026,67(09):985-993.
王楚越,李家诚,杨应奇等.通便汤对慢传输型便秘模型大鼠结肠组织VAPB-PTPIP51复合物及Cajal间质细胞自噬的影响[J].中医杂志,2026,67(09):985-993. DOI: 10.13288/j.11-2166/r.2026.09.011.
WANG Chuyue,LI Jiacheng,YANG Yingqi,et al.Effect of Tongbian Decoction (通便汤) on the VAPB-PTPIP51 Complex and Autophagy of Interstitial Cells of Cajal in the Colon of Slow Transit Constipation Model Rats[J].Journal of Traditional Chinese Medicine,2026,67(09):985-993. DOI: 10.13288/j.11-2166/r.2026.09.011.
目的
2
探讨通便汤治疗慢传输型便秘(STC)的可能作用机制。
方法
2
将24只SD大鼠随机分为正常组、模型组、通便汤组及莫沙必利组,每组6只。除正常组以外,其余各组均采用盐酸洛哌丁胺联合生理盐水灌胃法建立STC大鼠模型。造模成功后次日通便汤组大鼠给予通便汤药液18.63 g·kg
-1
灌胃,莫沙必利组大鼠给予每只1.605 mg·d
-1
灌胃,正常组和模型组大鼠给予10 ml·kg
-1
生理盐水灌胃,各组均每日灌胃1次,持续7天。最后一次灌胃24 h后计算各组大鼠粪便颗粒数、粪便含水率;10%活性炭悬溶液灌胃,30 min后计算小肠推进率;ELISA法检测血清中胃泌素(GAS)、胃动素(MTL)浓度;HE染色观察结肠组织病理形态,阿利新蓝-过碘酸-雪夫(AB-PAS)染色观察结肠黏液分泌情况;透射电镜观察结肠组织超微结构;Western Blot法检测结肠组织中干细胞因子受体(C-kit)、干细胞因子(SCF)、自噬相关蛋白5(ATG5)、Beclin1、囊泡相关膜蛋白B(VAPB)、蛋白酪氨酸磷酸酶相互作用蛋白51(VAPB-PTPIP51)蛋白水平;Real-time PCR法检测结肠组织中
C
-
kit
、
SCF
、
ATG5
、
Beclin1
、
VAPB、PTPIP51
mRNA水平,免疫组化检测结肠组织中SCF、C-kit、Beclin1、ATG5表达;ELISA法检测结肠组织钙离子(Ca
2+
)含量。
结果
2
与正常组比较,模型组大鼠粪便颗粒数、粪便含水率明显下降,小肠推进率降低,血清GAS、MTL浓度降低(
P
<0.01);肠道杯状细胞数量下降,肠黏膜层、肌层变薄;结肠组织中
ATG5、Beclin1
mRNA表达及其蛋白水平明显升高,Ca
2+
含量降低(
P
<0.05或
P
<0.01);电镜显示结肠细胞内空泡样变性,自噬体增多。与模型组比较,通便汤组和莫沙必利组大鼠粪便颗粒数、粪便含水率、小肠推进率增加,血清GAS、MTL浓度升高,结肠组织Ca
2+
含量升高,Beclin1、ATG5蛋白水平降低(
P
<0.05或
P
<0.01);黏膜层厚度及杯状细胞数量显著增加,结肠细胞内自噬体减少;通便汤组
ATG5、Beclin1
mRNA表达降低,莫沙必利组
SCF、VAPB、PTPIP51
mRNA表达升高,
Beclin1
mRNA表达降低(
P
<0.05或
P
<0.01)。与莫沙必利组比较,通便汤粪便颗粒数、粪便含水率、血清GAS浓度、结肠组织Ca
2+
含量、C-kit水平升高,ATG5、Beclin1水平降低(
P
<0.05或
P
<0.01)。
结论
2
通便汤可能通过上调线粒体-内质网互作时VAPB-PTPIP51复合物表达,减轻Cajal间质细胞自噬,促进肠道动力,从而改善便秘症状。
Objective
2
To explore the possible mechanism of
Tongbian Decoction
(通便汤, TD) in treating slow transit constipation (STC).
Methods
2
Twenty-four SD rats were randomly divided into normal group, model group, TD group, and mosapride group, with 6 rats per group. Except for the normal group, STC models were established by intragastric administration of loperamide hydrochloride combined with normal saline. On the day following successful model establishment, rats in the TD group received 18.63 g·kg⁻¹ of TD by gavage, while those in the mosapride group received 1.605 mg·d⁻¹ of mosapride, and those in the normal group and the model group received 10 ml·kg⁻¹ of normal saline by gavage. All treatments were administered once daily for 7 consecutive days. Twenty-four hours after the last administration, fecal pellet number and fecal water content were measured. After intragastric administration of a 10% activated charcoal suspension, the small intestinal transit rate was calculated 30 minutes later. Serum levels of gastrin (GAS) and motilin (MTL) were measured by ELISA. Colonic histopathology was observed by HE staining, and mucus secretion by Alcian blue-periodic acid-Schiff (AB-PAS) staining. Ultrastructure of colon tissue was examined using transmission electron microscopy. Protein expression levels of C-kit, stem cell factor (SCF), autophagy-related protein 5 (ATG5), Beclin1, vesicle-associated membrane protein B (VAPB), and protein tyrosine phosphatase interacting protein 51 (VAPB-PTPIP51) were measured by Western Blot, and the mRNA levels were detected by real-time PCR. Immunohistochemistry was used to detect SCF, C-kit, Beclin1, and ATG5 expression. The calcium content in colon tissue was determined by ELISA.
Results
2
Compared to the normal group, rats in the model group showed significantly reduced fecal pellet number, fecal water content, small in
testinal transit rate, and serum GAS and MTL levels (
P
<0.01); the number of goblet cells decreased, and the mucosal and muscular layers of the colon became thinner; mRNA and protein expression levels of ATG5 and Beclin1 in colon tissue significantly increased, while calcium content decreased (
P
<0.05 or
P
<0.01); and electron microscopy revealed vacuolar degeneration and increased autophagosomes in colonic cells. Compared to the model group, both TD group and mosapride group showed increased fecal pellet number, fecal water content, small intestinal transit rate, serum GAS and MTL levels, and colonic calcium content, along with decreased Beclin1 and ATG5 protein levels (
P
<0.05 or
P
<0.01); the mucosal thickness and goblet cell number increased significantly, and autophagosomes decreased; in the TD group, ATG5 and Beclin1 mRNA levels decreased; in the mosapride group, SCF, VAPB, and PTPIP51 mRNA levels increased, while Beclin1 mRNA decreased (
P
<0.05 or
P
<0.01). Compared to the mosapride group, the TD group showed higher fecal pellet number, fecal water content, serum GAS levels, colonic calcium content, and C-kit expression, along with lower ATG5 and Beclin1 levels (
P
<0.05 or
P
<0.01).
Conclusion
2
TD may improve constipation symptoms by upregulating the VAPB-PTPIP51 complex during mitochondria-endoplasmic reticulum interactions, reducing autophagy of interstitial cells of Cajal, and promoting intestinal motility.
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