化瘀通便汤调节STC大鼠结肠Cajal间质细胞自噬相关蛋白表达的实验研究  

Experimental Study on the Regulation of Autophagy-related Protein Expression in Colon Cajal Mesenchymal Cells of STC Rats by Huayu Tongbian Decoction(化瘀通便汤)

作　　者：满如 于永铎[2] 陈萌[3] 张斯瑶[3] 姜洪宇 MAN Ru;YU Yongduo;CHEN Meng;ZHANG Siyao;JIANG Hongyu(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China;Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110032,Liaoning,China;Third Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110003,Liaoning,China)

机构地区：[1]辽宁中医药大学,辽宁沈阳110847 [2]辽宁中医药大学附属医院,辽宁沈阳110032 [3]辽宁中医药大学附属第三医院,辽宁沈阳110003

出　　处：《辽宁中医药大学学报》2022年第6期23-28,共6页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：国家自然科学基金青年科学基金(81804094);辽宁省自然科学基金(2019-MS-228)。

摘　　要：目的该实验通过观察复方地芬诺酯诱导的慢传输型便秘(slow transit constipation,STC)模型大鼠结肠组织中Cajal间质细胞(ICCs)自噬情况,探讨化瘀通便汤治疗STC机制及其量效关系。方法选择成年72只SD大鼠,随机分为6组,空白对照组12只大鼠给予纯净水灌胃,剩余5组用复方地芬诺酯灌胃制备慢传输型便秘大鼠模型,造模2周后随机抽取1组实验组,和空白组一起收集粪便,采用检测粪便含水率和炭末推进率的方法进行肠道功能障碍的测定。确定造模成功后除模型对照组外,3组分别给予低、中、高剂量化瘀通便汤治疗,1组给予西药治疗,模型组予等量纯水灌胃,对6组大鼠各项指标进行观察,包括大鼠一般情况(精神状况、进食量、饮水量、首次排黑便时间),利用透射电镜观察ICCs形态变化,并使用免疫组化法、免疫荧光、蛋白免疫印迹法检测ICCs及自噬相关蛋白Beclin1、Atg5、LC3B、p62的表达。结果(1)与空白组相比,造模组大鼠一般情况较差,首次排黑便时间延长(P<0.01)。(2)相较于空白组,模型组Cajal间质细胞出现严重损伤,各干预组Cajal间质细胞的损伤均有所改善和修复,化瘀通便汤高剂量组Cajal间质细胞改善和修复最明显。(3)与空白组相比,模型组ICCs细胞结构明显改变,Beclin1、LC3B、Atg5蛋白表达上升,p62表达下降(P<0.01);与模型组相比,低、中、高剂量化瘀通便汤组自噬相关蛋白表达均有不同程度的变化,差异有明显统计学意义(P<0.01);高剂量化瘀通便汤与低、中组间差异显著(P<0.05)。结论(1)运用复方地芬诺酯成功复制STC大鼠模型,且STC大鼠模型稳定性好,安全性高,可用于后期实验研究。(2)化瘀通便汤能明显改善STC症状,修复Cajal间质细胞结构,主要机制可能与下调Beclin1、LC3B、Atg5表达而调节ICCs过度自噬,上调p62表达促进ICCs修复有关,从而能刺激肠道蠕动,增加肠道水分。(3)化瘀通便汤治疗SObjective To observe the autophagy of Cajal interstitial cells(ICCs)in colon tissue of slow-transit constipation(STC)model rats induced by compound diphenoxylate,and to explore the mechanism of Huayu Tongbian Decoction(化瘀通便汤)in treating STC and its dose-effect relationship.Methods Seventy-two adult SD rats were randomly divided into 6 groups,12 rats in the blank control group were gavaged with pure water,and the remaining 5 groups were gavaged with compound difenoxanthine to prepare a slow-transit constipation rat model.4 weeks after modeling,one experimental group was randomly selected,and feces were collected together with the blank group,and intestinal dysfunction was measured by detecting the fecal water content and carbon end propulsion rate.After the successful modeling,except for the model control group,three groups were treated with low,medium and high doses of Huayu Tongbian Decoction,one group was treated with Western medicine,and the model group was given an equal amount of pure water by gavage.The rats in the six groups were observed for various indexes,including general conditions(mental status,food intake,water intake,time of first black stool),and morphological changes of ICCs were observed using transmission electron microscopy,and immunohistochemistry,immunofluorescence,and protein immunoblotting were used.Immunofluorescence and protein immunoblotting were used to detect the expression of ICCs and autophagy-related proteins Beclin1,Atg5,LC3B and p62.Results(1)Compared with the blank group,the general condition of the rats in the modeling group was worse,and the time to first black stool was prolonged(P<0.01).(2)Compared with the blank group,the Cajal mesenchymal cells in the model group showed serious damage,and the damage of Cajal mesenchymal cells in each intervention group was improved and repaired,and the improvement and repair of Cajal mesenchymal cells in the high-dose group of Huayu Tongbian Decoction was the most obvious.(3)Compared with the blank group,the cell structure of ICC

关 键 词：慢性传输型便秘 化瘀通便汤 CAJAL间质细胞 自噬 中药量效关系 

分 类 号：R574.62[医药卫生—消化系统]