Screening and identification of proteins mediating senna induced gastrointestinal motility enhancement in mouse colon.
OBJECTIVE
To isolate the proteins involved in pharmacologic action of senna extract (SE) from mouse gastrointestinal tract and to explore the molecular mechanism of gastrointestinal motility change induced by SE.


METHODS
SE was administrated to mice by different routes. Gastrointestinal motility of mice was observed using cathartic, gastrointestinal propellant movement experiments and X-ray analysis. Mouse model for gastrointestinal motility enhancement was established through continuous gastric administration of SE at progressively increased dose. At 3 h and week 3, 4, 6 and 10, morphological changes of gastrointestinal tissues were found under light microscope. Ultrastructural changes of intestinal and colonic tissues at week 6 were observed under transmission electron microscope. The colonic proteomic changes in model mice were examined by two-dimension polyacrylamide gel electrophoresis with immobilized pH gradient isoelectric focusing to screen the differentially expressed proteins, and their molecular masses and isoelectric points were determined. Two N-terminal sequences of the samples were also determined by mass spectrometry.


RESULTS
SE (0.3g) caused diarrhea after gastric administration in 1-6h and enhanced gastrointestinal propellant (65.1+/-7.5%; 45.8+/-14.6%, P<0.01) in mice, but intramuscular and hypodermic injection had no cathartic effect. X-ray analysis of gastrointestinal motility demonstrated that gastric administration of SE enhanced gastric evacuation and gastrointestinal transferring function. At 3 h and week 3 and 4 after gastric administration of SE, light microscopic examination revealed no apparent change in gastrointestinal mucosal tissues, but transmission electron microscopic examination revealed inflammatory changes in whole layer of intestinal and colonic wall. Twenty differential proteins were detected in the colonic tissues of the model mice by two-dimensional electrophoresis, and the N-terminal amino acid sequences of two proteins were determined.


CONCLUSIONS
SE causes diarrhea and enhances gastrointestinal motility through digestive tract administration. Long-term gastric administration of SE induces inflammatory changes and cell damage in the whole gastrointestinal tract. The differential proteins screened from the colonic tissues of the model mice might mediate the enhancing effect of SE on gastrointestinal motility.
Wang
Xin
X
Institute of Digestive Diseases, Xijing Hospital, Fourth Military Medical University,Xi'an 710033,Shaanxi Province,China.
Zhong
Yue-Xia
YX
Lan
Mei
M
Zhang
Zong-You
ZY
Shi
Yong-Quan
YQ
Lu
Ju
J
Ding
Jie
J
Wu
Kai-Cun
KC
Jin
Jian-Ping
JP
Pan
Bo-Rong
BR
Fan
Dai-Min
DM
D000818:Animals; D002400:Cathartics / Q000494:pharmacology; D003106:Colon / Q000737:chemistry / Q000187:drug effects / Q000502:physiology*; D003967:Diarrhea / Q000139:chemically induced* / Q000000981:diagnostic imaging / Q000503:physiopathology; D004195:Disease Models, Animal; D015180:Electrophoresis, Gel, Two-Dimensional; D004750:Enteral Nutrition; D005260:Female; D005769:Gastrointestinal Motility / Q000187:drug effects*; D007279:Injections, Subcutaneous; D007413:Intestinal Mucosa / Q000737:chemistry / Q000187:drug effects / Q000648:ultrastructure; D007525:Isoelectric Focusing; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D010936:Plant Extracts / Q000494:pharmacology*; D011506:Proteins / Q000032:analysis*; D011859:Radiography; D029846:Senna Plant
D002400:Cathartics; D010936:Plant Extracts; D011506:Proteins