肾小管上皮细胞表达三叶因子1抑制草酸钙结石形成的机制研究  

Mechanism of inhibiting calcium oxalate stone formation by expression of trefoil factor 1 in renal tubular epithelial cells

作　　者：周治彦 符红霞 陈金火 孙早喜 Zhou Zhiyan;Fu Hongxia;Chen Jinhuo;Sun Zaoxi(Department of Urology,First Afiliated Hospital,Hainan Medical University,Haikou 570102,China;Department of Hepatobiliary Surgery,First Affiliated Hospital,Hainan Medical University,Haikou 570102,China)

机构地区：[1]海南医科大学第一附属医院泌尿外科,海口570102 [2]海南医科大学第一附属医院肝胆外科,海口570102

出　　处：《中华实验外科杂志》2024年第10期2277-2280,共4页Chinese Journal of Experimental Surgery

基　　金：海南省自然科学基金(20158350)。

摘　　要：目的:研究三叶因子1(TFF1)在肾小管上皮细胞(RTECs)中表达及其抑制肾脏草酸钙(COM)结石成石的机制。方法:获取SPF级健康雄性大鼠(SD)RTECs并原代培养;RTECs分为TFF1表达组共6小组,COM不同浓度影响TFF1表达共4组:空白对照组、1 mmol/L组、2 mmol/L组和4 mmol/L组,COM最佳浓度(2 mmol/L)对TFF1 RNAi表达影响共3组:阴性对照组、TFF1RNAi组及其重复组。以免疫荧光法、Western blot法、免疫组织化学法检测、方差分析法及LSD检验分析RTECs中TFF1的表达。结果:TFF1均表达于RTECs,且主要表达于胞浆中,细胞核有少量表达。2 mmol/L浓度时胞质含大量大小不等棕褐色阳性颗粒,其它各浓度及空白对照时胞质含少量大小不等棕褐色阳性颗粒,并且2 mmol/L浓度时抑制RTECs生长,细胞形态较小。不同浓度COM共培养RTECs后,TFF1表达由高到低依次为实验组3、组4、组2及空白对照组1(P<0.01),其比例为1.00∶1.33∶2.00∶1.78。沉默TFF1表达后,选择2 mmol/L的COM共培养RTECs,TFF1的表达水平由高到低依次为组1、组2、组3(P<0.05),组2与组3比较其差异无统计学意义,其比例为1.00∶0.44∶0.45。结论:TFF1可能是肾草酸钙结石抑制剂,为草酸钙结石抑制的进一步研究提供理论依据。Objective:Study the expression of Trefoil factor 1(TFF1)in renal tubular epithelial cells and its mechanism in the formation of renal calcium oxalate stones.Methods:Obtain SPF grade healthy male rat(SD)RTECs and perform primary cell culture.RTECs were divided into 6 groups to validate TFF1 expression.Different concentrations of COM affect TFF1 expression in four groups:blank control group,1 mmol/L group,2 mmol/L group,and 4 mmol/L group.The optimal concentration of COM(2 mmol/L)affects the expression of TFF1 RNAi in three groups:negative control group,TFF1 RNAi group,and its repeat group.Analyze the expression of TFF1 in RTECs using immunofluorescence,western blot,immunohistochemistry,analysis of variance,and LSD test.Results:TFF1 is expressed in RTECs,mainly in the cytoplasm,with a small amount expressed in the nucleus.At a concentration of 2 mmol/L,the cytoplasm contains a large number of brown positive particles of varying sizes,while at other concentrations and blank controls,the cytoplasm contains a small amount of brown positive particles of varying sizes.And at a concentration of 2mmol/L,it inhibits the growth of RTECs and results in smaller cell morphology.After co culturing RTECs with different concentrations of COM,the expression of TFF1 was ranked from high to low as experimental group 3,group 4,group 2,and blank control group 1(P<0.01),with a ratio of 1.00∶1.33∶2.00∶1.78.After silencing TFF1 expression,2 mmol/L COM co cultured RTECs were selected.The expression levels of TFF1 were ranked from high to low in Group 1,Group 2,and Group 3(P<0.05).There was no statistically significant difference between Group 2 and Group 3,with a ratio of 1.00∶0.44∶0.45.Conclusion:TFF1 may be an inhibitor of renal calcium oxalate stones,providing a theoretical basis for further research on the inhibition of calcium oxalate stones.

关 键 词：三叶因子1 肾小管上皮细胞 草酸钙结石 成石机制 

分 类 号：R692.6[医药卫生—泌尿科学]