附子-肉桂配伍对阳虚型慢传输型便秘大鼠肠动力及SCF、c-Kit表达的影响

Effect of Aconiti Lateralis Radix Praeparata Combined with Cinnamomi Cortex on Intestinal Motility and Expression of SCF and c-Kit in Slow Transit Constipation Rats with Yang Deficiency Syndrome

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DOI：10.7661/j.cjim.20230828. 058

中文关键词: 慢传输型便秘附子-肉桂肠道动力神经递质 SCF、c-Kit信号通路中药

英文关键词:slow transit constipation Aconiti lateralis Radix Praeparata compined with Cinnamomi Cortex intestinal motility neurotransmitter SCF and c-Kit signaling pathway Chinese herbal medicine

基金项目:贵州省自然科学基金资助项目（No.黔科合基础[2020]1Y362， No.黔科合基础-ZK[2022]一般510）;贵州省中医药、民族医药科学技术研究专项课题项目（No.QZYY-2021-016）

作者	单位
张远哲;董文然;赵罗娜;杨元凤;黎豫川	1.贵州中医药大学基础医学院（贵阳 550025）,2.北京中医药大学药学院（北京 100029）,3.贵州中医药大学第二附属医院药学部（贵阳 550025）

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中文摘要:

目的研究附子-肉桂配伍对阳虚型慢传输型便秘（STC）大鼠肠神经递质、干细胞因子（SCF）及酪氨酸激酶（c-Kit）表达的影响，并探讨其改善肠动力的作用机制。方法将90只SD大鼠随机分为空白组（15只）和模型组（75只），采用盐酸洛哌丁胺结合0 ℃冰水复合灌胃法建立阳虚型STC大鼠模型。造模成功后70只模型大鼠随机分为模型对照组、附子-肉桂（2： 1）组、附子-肉桂（1： 2）组、附子-肉桂（1： 1）组、附子组、肉桂组、普芦卡必利组，每组10只，同时取空白组为10只。各中药组分别给予相应水煎液灌胃，普芦卡必利组给予0.18 mg/kg普芦卡必利混悬液灌胃，空白组和模型对照组给予10 mL/kg体积蒸馏水灌胃，每天1次，各组均连续灌胃14天。观察大鼠一般情况、粪便含水率、肠道推进率，ELISA法检测结肠5-羟色胺（5-HT）、血管活性肠多肽（VIP）、P物质（SP）含量；免疫组化法观察结肠5-HT、VIP、SP、SCF、c-Kit表达情况；Western Blot法测定SCF、c-Kit蛋白水平，RT-PCR法检测c-Kit、SCF mRNA表达水平。结果与空白组比较，模型对照组粪便含水率、肠道推进率、结肠5-HT、SP表达、SCF、c-Kit蛋白和mRNA表达均下降（P<0.01），结肠VIP表达增加（P<0.01）。与模型对照组比较，附子-肉桂（2： 1）组、普芦卡必利组粪便含水率、肠道推进率、结肠5-HT、SP表达、SCF、c-Kit蛋白和mRNA表达水平增高（P<0.01），结肠VIP表达降低（P<0.01）。与附子-肉桂（2： 1）组和普芦卡必利组比较，附子组、肉桂组c-Kit、SCF 蛋白和mRNA表达水平降低（P<0.05，P<0.01）。结论附子-肉桂（2： 1）配伍可有效改善阳虚型STC肠道动力障碍，其促进肠动力的作用与下调VIP表达、上调结肠5-HT、SP、SCF、c-Kit表达水平有关。

英文摘要:

Objective To investigate the effects of the Aconiti lateralis Radix Praeparata-Cinnamomi Cortex（ALRP-CC）compound on the expression of intestinal neurotransmitters and stem cell factor（SCF），c-kitproto-oncogeneprotein （c-Kit） in slow transit constipation（STC）rats with Yang deficiency syndrome， and to explore its mechanism of action for improving intestinal motility. Methods Totally 90 SD rats were randomly divided into the blank group （n=15） and model group （n=75）. STC rats with Yang deficiency syndrome were established by loperamide hydrochloride in ice water at 0 ℃ intragastrically. After successful modeling，70 rats were randomly divided into the model control group， ALRP-CC（2： 1）group， ALRP-CC（1： 2）group， ALRP-CC（1： 1） group， ALRP group， CC group， and prucalopride group， 10 in each group， and the blank group including 10 rats. Rats in each Chinese medicine group were given the corresponding water decoction intragastrically；rats in the prucalopride group were given prucalopride suspension 0.18 mg·kg-1 intragastrically. Meanwhile， rats in the blank group and model control group were given distilled water 10 mL·kg-1 intragastrically， once daily for 14 consecutive days. Rats' general condition， water content in feces， and intestinal transit rate were observed. The 5-hydroxy-tryptamine （5-HT）， vasoactive intestinal polypeptide （VIP）， and substance P（SP） in the colon were detected using ELISA. The expression of 5-HT， VIP， SP， SCF and c-Kit in the colon were observed by immunohistochemistry. The expression of SCF and c-Kit proteins and mRNA were detected by Western Blot and RT-PCR， respectively. Results Compared with the blank group， the water content in feces， intestinal transit rate， 5-HT and SP expression as well as SCF and c-Kit protein and mRNA expression in the colon decreased （all P<0.01）， and VIP expression in the colon decreased（P<0.01）in the model control group. Compared with the model control group， the water content in feces， intestinal transit rate， and 5-HT and SP expression， as well as SCF and c-Kit protein and mRNA expression in the colon increased（P<0.01）， and VIP expression in the colon decreased （P<0.01）in the ALRP-CC（2： 1） group and prucalopride group. Compared with the ALRP-CC（2： 1）group and prucalopride group， expression of c-Kit and SCF protein and mRNA decreased in ALRP group and CC group （P<0.05， P<0.01）. Conclusions ALRP-CC（2： 1）could effectively improved intestinal motility disorder. And its mechanism might related to down-regulated VIP expression and up-regulated 5-HT， SP ，SCF and c-Kit expression in the colon.

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