黄育生,刘媛,钟如帆,唐洪梅,王婷,石玉莹,祝赫
HUANG Yusheng,LIU Yuan,ZHONG Rufan,TANG Hongmei,WANG Ting,SHI Yuying,ZHU He

• 1:广州中医药大学
• 2:广州中医药大学第一附属医院

摘要(Abstract)：

目的探讨JAK2/STAT3信号通路在大鼠海马中的表达以及肠激安方改善腹泻型肠易激综合征(IBS-D)的作用机制。方法采用母婴分离、醋酸刺激及束缚应激结合的方法建立IBS-D大鼠模型;造模成功后将大鼠随机分为模型对照组、匹维溴铵组(0.018 g·kg~(-1))、肠激安方高剂量组(33.48 g·kg~(-1))、肠激安方低剂量组(16.74 g·kg~(-1)),另取同批次正常大鼠作为正常对照组,各6只,连续灌胃14 d,正常对照组和模型对照组给予等体积生理盐水;给药结束后取海马组织,Q-PCR检测JAK2、STAT3、TNF-α、IL-6的基因表达,免疫印迹(Western Blot)法检测JAK2、STAT3、pSTAT3蛋白表达。结果与正常对照组比较,IBS-D大鼠海马中JAK2、STAT3、TNF-αmRNA及JAK2、STAT3、pSTAT3蛋白表达明显增加,差异有统计学意义(P<0.05),IL-6mRNA虽有上升的趋势,但是差异无统计学意义(P>0.05);肠激安方高剂量组JAK2、STAT3、TNF-αmRNA及JAK2、STAT3、pSTAT3蛋白表达较模型组减少,差异有统计学意义(P<0.05);肠激安方低剂量组STAT3、TNF-αmRNA及JAK2、STAT3蛋白表达较模型组减少,差异有统计学意义(P<0.05),JAK2 mRNA及pSTAT3蛋白有减少的趋势,但差异无统计学意义(P>0.05)。匹维溴铵组STAT3 mRNA相对表达量较模型组减少,差异有统计学意义(P<0.05),TNF-α、JAK2 mRNA及JAK2、STAT3、pSTAT3蛋白表达有减少的趋势,但差异无统计学意义(P>0.05)。结论 IBS-D大鼠海马中JAK2/STAT3通路被激活,肠激安方可抑制该信号通路的活性,这可能是疏肝健脾法改善IBS-D症状的作用机制之一。
OBJECTIVE To investigate the expression of JAK2/STAT3 signaling pathway in the hippocampus of IBS-D rats and the mechanism of Changji' an prescription on improving IBS-D. METHODS IBS-D rat model was established by method of combination of separation of breast milk,stimulation of acetic acid and constraint of four limbs. All model rats were randomly divided into the model control group,the pinaverium bromide group( 0.018 g·kg~(-1)),the high dose of Changji' an prescription group( 33.48 g·kg~(-1)),and the low dose of Changji' an prescription group( 16. 74 g·kg~(-1)),six in each group. All medication lasted for 14 successive days.Normal saline was given to another six normal rats in the normal control group. The gene expression of JAK2,STAT3,TNF-α and IL-6 in hippocampus were detected by real-time fluorescent quantitative PCR. And the protein expression of JAK2,STAT3 and pSTAT3 were detected by Western Blot method. RESULTS Compared with the normal group,the mRNA expression of JAK2,STAT3 and TNF-α,and the protein expression of JAK2,STAT3 and pSTAT3 in the hippocampus of IBS-D rats was significantly increased( P<0.05),while IL-6 mRNA had an upward trend but there was no significant difference( P > 0. 05). While the mRNA expression of JAK2,STAT3 and TNF-α,and the protein expression of JAK2,STAT3 and pSTAT3 in the high dose of Changji' an prescription group was lower than that of model group( P<0.05).The mRNA expression of STAT3 and TNF-α,and the protein expression of JAK2 and STAT3 in the low dose of Changji'an prescription group was lower than that of model group( P < 0. 05) and that mRNA expression of JAK2 and the protein expression of pSTAT3 had a tendency to decrease,but the difference was not statistically significant( P>0.05).The mRNA expression of STAT3 in the pinaverium Bromide group was lower than that in the model group( P< 0.05),while the mRNA of expression of TNF-α,JAK2 and the protein expression of JAK2,STAT3 and pSTAT3 were decreased,but there was no significant difference( P > 0. 05).CONCLUSION The JAK2/STAT3 pathway is activated in the hippocampus of IBS-D rats,and the activity of the signaling pathway can be inhibited by Changji' an prescription,which may be the mechanism of the effect of soothing liver and strengthening spleen on improving the symptoms of IBS-D.

关键词(KeyWords)： 疏肝健脾;腹泻型肠易激综合征;JAK2/STAT3
soothing liver and strengthening spleen;IBS-D;JAK2/STAT3

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(81673842,81073160);; 广东省自然科学基金项目(2014A030313404)

作者(Author): 黄育生,刘媛,钟如帆,唐洪梅,王婷,石玉莹,祝赫
HUANG Yusheng,LIU Yuan,ZHONG Rufan,TANG Hongmei,WANG Ting,SHI Yuying,ZHU He

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