目的 探究消痔丸對復(fù)方地芬諾酯致小鼠便秘模型的藥效作用及機(jī)制。方法 ICR小鼠隨機(jī)分為對照組、模型組、溴化甲基納曲酮（陽性對照，58.5 mg·kg^-1）組和消痔丸低、中、高劑量（1.17、2.34、4.68 g·kg^-1）組，連續(xù)ig給藥7 d，給藥同時(shí)除對照組外，其余各組按照7.5 mg·kg^-1 ig復(fù)方地芬諾酯混懸液制備小鼠便秘模型。觀察小鼠一般情況；末次給藥結(jié)束后，各實(shí)驗(yàn)組禁食不禁水16 h，模型組和各給藥組ig給予復(fù)方地芬諾酯，30 min后給藥組ig含相應(yīng)受試藥的活性炭懸液，對照組、模型組ig活性炭懸液。第1批小鼠觀察首次排黑便時(shí)間、6 h內(nèi)排便粒數(shù)、6 h和24 h內(nèi)排便質(zhì)量及糞便含水率。第2批小鼠檢測小腸推進(jìn)率，酶聯(lián)免疫吸附實(shí)驗(yàn)（ELISA）檢測結(jié)腸組織和血清中5-羥色胺（5-HT）、P物質(zhì)（SP）、血管活性腸多肽（VIP）水平變化，實(shí)時(shí)熒光定量PCR（qRT-PCR）法檢測結(jié)腸組織水通道蛋白3（AQP3）、AQP9 mRNA水平，Westernblotting法檢測AQP3、AQP9蛋白表達(dá)水平。結(jié)果 與模型組相比，各給藥組小鼠糞便干結(jié)情況有所好轉(zhuǎn)，排便量均有不同程度的增加，體質(zhì)量有所增加，精神狀態(tài)逐漸好轉(zhuǎn)。與模型組相比，各給藥組均能顯著縮短排首粒黑便時(shí)間，提高6 h排便粒數(shù)及糞便含水率、6 h及24 h排便質(zhì)量（P＜0.05、0.01、0.001）；除消痔丸低劑量組，其余各給藥組小鼠24 h糞便含水率均顯著增加（P＜0.01、0.001）；各給藥組小腸炭末推進(jìn)距離及炭末推進(jìn)率均明顯提高（P＜0.05、0.01、0.001）；各給藥組均能顯著降低結(jié)腸組織中VIP水平（P＜0.01、0.001），除消痔丸低劑量組，其余各給藥組均能顯著提高結(jié)腸組織中5-HT、SP水平（P＜0.01、0.001）；各給藥組均能顯著增加血清中5-HT、SP水平并降低VIP的表達(dá)水平（P＜0.05、0.01、0.001）；各給藥組AQP3蛋白表達(dá)量均顯著下降（P＜0.01、0.001），AQP9蛋白表達(dá)量均顯著上升（P＜0.05、0.01、0.001）；各給藥組AQP3 mRNA水平均顯著下降（P＜0.01、0.001），AQP9 mRNA水平均顯著上升（P＜0.05、0.01、0.001）。結(jié)論 消痔丸對復(fù)方地芬諾酯誘導(dǎo)的小鼠便秘模型有顯著的治療效果，可以顯著縮短便秘小鼠排首粒黑便時(shí)間、增加排便粒數(shù)并提高糞便含水率、顯著促進(jìn)小腸推進(jìn)作用，作用機(jī)制與調(diào)節(jié)腸液分泌相關(guān)因子5-HT、SP、VIP等的釋放以及AQP3、AQP9表達(dá)相關(guān)。

Objective To investigate the therapeutic effect of Xiaozhi Pills (XZP) on constipation induced by diphenoxylate in mice and its mechanism. Methods ICR mice were randomly divided into control group, model group, methylnaltrexone bromide group (positive control, 58.5 mg·kg^-1) and XZP low, medium, and high doses group (1.17, 2.34, and 4.68 g·kg^-1). Each group was administered by gavage of the corresponding drug once a day for seven days. Except for the control group, the other groups were given diphenoxylate suspension at 7.5 mg·kg^-1 body weight by gavage on the same day. Observe the general situation of mice. After the end of the last administration, each experimental group fasted for 16 hours and couldn't help but water. The model group and each administration group were given compound diphenoxylate by ig. After 30 minutes, the administration group ig contained activated carbon suspension of the corresponding test drug, while the control group and model group ig contained activated carbon suspension. The first batch of mice were observed for the first time of black stools, the number of stools within six hours, the quality of stools within 6 and 24 hours, and the fecal moisture content. The second batch of mice were tested for intestinal propulsion rate, enzyme-linked immunosorbent assay (ELISA) was used to detect changes in levels of serotonin (5-HT), substance P (SP), and vasoactive intestinal polypeptide (VIP) in colon tissue and serum, real-time fluorescence quantitative PCR (qRT-PCR) was used to detect levels of aquaporin 3 (AQP3) and AQP9 mRNA in colon tissue, and Western blotting was used to detect protein expression levels of AQP3 and AQP9. Results Compared with the model group, the fecal dryness of mice in each treatment group improved, with varying degrees of increase in fecal volume, increased body mass, and gradually improved mental state. Compared with the model group, each medication group significantly shortened the first black stool discharge time, improved the number of fecal particles and fecal moisture content at six hours, and improved the quality of feces at 6 and 24 hours (P < 0.05, 0.01, 0.001); Except for the low-dose group of XZP, the 24-hour fecal moisture content of mice in all other treatment groups significantly increased (P < 0.01, 0.001). The distance and rate of small intestine charcoal powder propulsion in each administration group were significantly increased (P < 0.05, 0.01, 0.001). All treatment groups were able to significantly reduce VIP levels in colon tissue (P < 0.01, 0.001). Except for the low-dose group of XZW, all other treatment groups were able to significantly increase the levels of 5-HT and SP in colon tissue (P < 0.01, 0.001). Each administration group significantly increased the levels of 5-HT and SP in serum and decreased the expression level of VIP (P < 0.05, 0.01, 0.001). The expression of AQP3 protein significantly decreased in each treatment group (P < 0.01, 0.001), while the expression of AQP9 protein significantly increased (P < 0.05, 0.01, 0.001). The levels of AQP3 mRNA was significantly decreased in each treatment group (P < 0.01, 0.001), while the levels of AQP9 mRNA was significantly increased (P < 0.05, 0.01, 0.001). Conclusion XZP had a significant therapeutic effect on the constipation model induced by compound diphenoxylate in mice. It can significantly shorten the first black stool time, increase the number of stool particles and increase the fecal water content, and significantly promote small intestine propulsion. The mechanism of action is related to regulating the release of intestinal secretion related factors such as 5-HT, SP, VIP, and the expression of AQP3 and AQP9.

R285.5

國家自然科學(xué)基金重點(diǎn)項(xiàng)目（U21A20406）；蘭州市人才創(chuàng)新創(chuàng)業(yè)項(xiàng)目（2022-RC-2）