基于“脑-肠轴”线粒体未折叠蛋白反应研究济川煎对慢传输型便秘的作用机制  

Mechanism of Jichuanjian for Treatment of Slow Transit Constipation Based on Brain-Gut Axis Mitochondrial Unfolded Protein Reaction

作　　者：梁星琛 白璐璐 周永学[1] 闫曙光[1] 周小燕 LIANG Xing-chen;BAI Lu-lu;ZHOU Yong-xue;YAN Shu-guang;ZHOU Xiao-yan(Basic Medcine College,Shaanxi University of Chinese Medcine,Shaanxi(712046);Tumor Department,Affiliated Hospital,Shaanxi University of Chinese Medcine,Shaanxi(712000))

机构地区：[1]陕西中医药大学基础医学院,陕西712046 [2]陕西中医药大学附属医院肿瘤科,陕西712000

出　　处：《中国中西医结合杂志》2022年第1期75-82,共8页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(No.81273663);陕西中医药大学校级科研课题(No.2020CX38)。

摘　　要：目的观察济川煎对慢传输型便秘(STC)大鼠海马神经元与结肠细胞线粒体未折叠蛋白反应(UPRmt)水平的影响,结合"脑-肠轴"探讨补肾益精法治疗STC的作用机制。方法 60只清洁级SD大鼠按随机数字法抽取10只为空白组,余采用复方地芬诺酯(15 mg/kg,1次/日)灌胃6周复制STC模型,造模成功后大鼠分为:模型组、济川煎低、中、高剂量组、阳性药物组各10只。造模成功后空白组、模型组采用生理盐水等体积灌胃,济川煎低(8.64 g/kg,1次/日)、中(17.28 g/kg,1次/日)、高(34.56 g/kg,1次/日)剂量组、阳性药物组(莫沙必利1.54 mg/kg,1次/日)予以相应药物灌胃2周。观察各组大鼠首粒黑便排出时间;取结肠组织HE、AB-PAS染色观察病理学变化;取海马组织尼氏染色观察尼氏小体变化;采用实时聚合酶链式反应(RT-PCR)、免疫荧光法以及Western Blot检测结肠、海马组织中HSP60、CLpP、CHOP、LON mRNA及蛋白表达水平。结果与空白组比较,模型组大鼠首粒黑便排出时间延长(P<0.01),结肠组织肌层纤维断裂,肌细胞核固缩、深染,结缔组织疏松,黏膜杯状细胞萎缩,管腔内黏液大量减少,海马组织中神经元数量减少,结肠组织与海马组织中HSP60、CLpP、CHOP、LON mRNA相对表达与蛋白表达均显著升高(P<0.01);与模型组比较,各治疗组结肠组织肌层纤维断裂、坏死减轻,结缔组织紧密,胞核饱满且无固缩,黏膜杯状细胞及黏液分泌有不同程度恢复,海马组织中神经元数量明显增多。各给药组结肠组织中HSP60、CLpP、CHOP和LON mRNA相对表达显著降低(P<0.01);海马组织中HSP60、CLpP、CHOP和LON mRNA相对表达有不同程度的降低(P<0.01,P<0.05);结肠组织中HSP60、CLpP、CHOP和LON蛋白表达均显著降低(P<0.01)。结论济川煎治疗STC的作用机制可能与调控结肠细胞、海马神经元的UPRmt以改善线粒体功能,通过脑肠同治减轻结肠细胞的凋亡与衰老,修复肠道肠道平滑肌�Objective To observe the effect of Jichuanjian(JCJ)on the levels of hippocampal neuron UPRmt and colon cell UPRmt in rats with slow transit constipation(STC), and to study the mechanism of treating STC by Shen tonifying essence replenishing method combined with "brain-gut" axis. Methods Ten rats were selected from 60 clean grade SD rats by random digit table, STC models were duplicated in the rest rats by intragastrically administered with Compound diphenoxylate(15 mg/kg, once per day). After successful modeling, the modeled rats were divided into model group,low dose JCJ group(8.64 g/kg), medium dose JCJ group(17.28 g/kg), high dose JCJ group(34.56 g/kg), and positive controlgroup(Mosapride, 1.54 mg/kg), 10 in each group. Equal volum of normal saline was administered to rats in the blank group and the model group All medication was performed once per day for 2 successive weeks. The first black fecal passing time was observed in each group. The pathological changes of the colon tissue were observed by HE staining an AB-PAS. The changes of Nissl bodies were observed by taking the hippocampal tissue for Nissl staining. RT-PCR, immunofluorescence, and Western Blot were used to detect mRNA and protein expression levels of HSP60, CLpP, CHOP, and LON in the colon tissue and hippocampus. Results Compared with the blank group, the discharge time of the first black stool was significantly prolonged in the model group(P<0.01). HE staining showed muscle fiber rupture, necrosis, pyknosis and deep staining of muscle cell nuclei, loose connective tissue, and AB-PAS staining showed mucosal goblet cell atrophy, and mucus in the lumen greatly decreased. In the hippocampus, the number of Nissl bodies decreased significantly. Western Blot and mRNA test results showed HSP60, CLpP, CHOP, LON mRNA and protein expressions significantly increased in the colon tissue and the hippocampus(P<0.01). Compared with the model group, muscular fiber breakage, reduced necrosis, tight connective tissue, full nucleus and no pyknosis, mucosal goblet

关 键 词：济川煎 慢传输型便秘 线粒体未折叠蛋白反应 脑-肠轴 

分 类 号：R285[医药卫生—中药学]