腹部推拿对非酒精性脂肪肝病大鼠肠上皮细胞MLCK、P-MLC表达及P-MLC与F-actin共定位的影响

Effect of abdominal tuina on the expression of MLCK,P-MLC and co localization of P-MLC and F-actin in intestinal epithelial cells in rats with non-alcoholic fatty liver disease

关键词:  腹部推拿  非酒精性脂肪肝病  肠道黏膜通透性  细胞骨架

Key words:  abdominal tuina  non-alcoholic fatty liver disease  intestinal mucosal permeability  cytoskeleton

摘要:

[目的] 研究腹部推拿对非酒精性脂肪肝病（NAFLD）大鼠肠上皮细胞肌球蛋白轻链激酶（MLCK）、肌球蛋白轻链磷酸化（P-MLC）表达及P-MLC与多聚体的纤维状肌动蛋白（F-actin）共定位的影响。[方法] 复制NAFLD大鼠模型，采用随机数字表法分为腹部推拿组、模型组，每组9只。另设9只大鼠为正常对照组。腹部推拿组每日进行10 min推拿干预；模型组每日束缚大鼠，10 min后解开束缚。正常对照组不予任何干预手段；干预28 d后，利用免疫蛋白印迹（Western blot）方法检测MLCK及P-MLC蛋白表达；同时利用免疫荧光方法检测肠黏膜P-MLC与F-actin的共定位关系。[结果] 造模后MLCK及P-MLC的表达明显上升。而经过腹部推拿干预后，MLCK及P-MLC的表达下降，证明腹部推拿可以抑制MLCK及P-MLC的表达；同时，免疫荧光方法检测肠黏膜P-MLC与F-actin之间存在共定位，且腹部推拿干预的P-MLC和F-actin的表达量最高，证明腹部推拿可以促进P-MLC和F-actin共定位作用。[结论] 腹部推拿可以通过抑制MLCK及P-MLC蛋白表达及促进P-MLC和F-actin相互作用的途径重塑肠上皮细胞骨架，改善NAFLD大鼠肠道黏膜通透性，逆转NAFLD的脂肪变性。

Abstract:

[Objective] To study the effect of abdominal tuina on the expression of MLCK,P-MLC and co localization of P-MLC and F-actin in intestinal epithelial cells in rats with non-alcoholic fatty liver disease.[Methods] NAFLD rat models were induced by high fat diet. The rats were divided into abdominal tuina group and model group by random number table,with 9 rats in each group. Another 9 rats were set as normal control group. The abdominal tuina group was given tuina intervention every day for 10 minutes,while the model group was confined to the experimental platform every day and took supine position without any intervention. The rats were released after 10 minutes. After 28 days of continuous intervention,the protein expression of MLCK and P-MLC were detected by Western blot;The co localization of P-MLC and F-actin in intestinal mucosa were detected by immunofluorescence.[Results] The expression of MLCK and P-MLC increased significantly after modeling;After abdominal Tuina intervention,the expression of MLCK and P-MLC decreased,which proved that abdominal tuina could inhibit the expression of MLCK and P-MLC;Immunofluorescence method showed that there was co localization between P-MLC and F-actin in intestinal mucosa,and the expression of P-MLC and F-actin in abdominal massage intervention were the highest,which proved that abdominal massage could promote the interaction between P-MLC and F-actin.[Conclusion] Abdominal tuina can remodel the cytoskeleton of intestinal epithelium by inhibiting the expression of MLCK and P-MLC protein and promoting the interaction of P-MLC and F-actin,so as to improve the intestinal mucosal permeability and reverse the fatty degeneration of NAFLD rats.