S Pain And Withdrawal Of D - Ibs Model Rats Colon Tissue Aqp3 Expression

Purpose:To observe effect on the expression of Tong Xie Yao Fang (Important Formula for Frigid Extremities Decoction) and its combining formula to colon tissue aquaporin3(AQP3) of diarrhea irritable bowel syndrome rat models and discuss prescribing meaning of tonifying spleen, nourishing liver and regulating qi in this formula according to ways of morphology, molecular biology and immunohistochemistry.Method:To divide60SPF Wistar male rats into six groups randomly:control group, model group, Tong Xie Yao Fang group, Tong Xie Yao Fang group without bai zhu group, Tong Xie Yao Fang group without bai shao group and Tong Xie Yao Fang group without chen pi group, and then feed them in six cages with ten in each one. We let rats receive six kinds of chronic stress stimulation:swimming in cold water, clamping trails, banning to drink water, banning to eat food, electric shock and bottoming up day and night, meanwhile, tie their bodies with made-to-order iron cages in order to set up irritable bowel syndrome (D-IBS) models every day. Next, we observe changes of rats in common conditions and identify models according to model estimating standard, and then the model typed successfully. To treat rats separately with Chinese drug liquids of Tong Xie Yao Fang and its combining formula being equivalent to ten times of adult dose for ten days and one time each day. After collecting specimen, detect colon tissue aquaporin3expressions of various rat models with immunohistochemistry, detect and analyze colon tissue aquaporin3mRNA expression of various rat models with revorse transcriptase polymerase chain reaction (RT-PCR) method. Experimental data adopt SPSS13.0software to analyze.Result: 1. Common states observation:Comparing with control group, animals in Model group appear dried-up fur, drooping spirit, slim physique, huddling, lassitude, soft bowel, dung adhered to anus, etc. Comparing with model group, diarrhea states of animals in groups without bai zhu and without bai shao have no obvious changes, and state of the group without chen pi is a little better such as sheen fur, increased weight and soft but figured bowel, while state of the Tong Xie Yao Fang group has obvious improvement, like more sheen fur, lustrous eyes, increased weight, sensitive action, figured bowel and recuperative diarrhea.2. Percentage of soft bowel:Percentage of model group is higher than that of control group. there is no significant difference for groups without bai zhu and without bai shao in contrast with model group. Percentage of group without chen pi is reduced (P<0.05). Percentage of Tong Xie Yao Fang group is notably decreased (P<0.01)3. Level of soft bowel:Level of model group is higher than that of control group. There is no significant difference for groups without bai zhu and without bai shao in contrast with model group. Level of group without chen pi is reduced (P<0.05). Level of Tong Xie Yao Fang group is notably decreased (P<0.01)4. Diarrhea index:Diarrhea index of the control group is0.00+0.00. There is no significant difference for groups without bai zhu and without bai shao in contrast with model group (P>0.05). Diarrhea index of group without chen pi is reduced (P<0,05). Diarrhea index of Tong Xie Yao Fang group is notably decreased.5. Immunohistochemistry detection show:AQP3protein expression in colon tissue mucosa layer of model group is obviously reduced (P<0.01) in contrast with control group. There is no significant difference for groups without bai zhu and without bai shao in contrast with model group (P>0.05). That of group without chen pi is increased (P<0.01). That of Tong Xie Yao Fang group is notably increased (P<0.01)8. According to β-actin as the internal control, AQP3mRNA protein expression in colon tissue mucosa layer of model group is obviously reduced (P<0.01) in contrast with control group. There is no significant difference for groups without bai zhu and without bai shao in contrast with model group (P>0.05) That of group without chen pi is increased (P<0.01). That of Tong Xie Yao Fang group is notably increased (P<0.01)Conclusion:1. Tong Xie Yao Fang group and group without chen pi can notably decrease percentage of soft bowel, level of soft bowel and Diarrhea index. There is no notable affection for groups without bai zhu and without bai shao in contrast with the above.2. Tong Xie Yao Fang group and group without chen pi can notably increase AQP3protein expression in colon tissue, while there is no notable affection for groups without bai zhu and without bai shao in contrast with the above.3. Tong Xie Yao Fang group and group without chen pi can notably increase AQP3mRNA protein expression in colon tissue, while there is no notable affection for groups without bai zhu and without bai shao in contrast with the above.