姜黄素对肠上皮细胞屏障损伤保护作用的研究

王改1*王娜2郝京霞3

保定市儿童医院PICU1(071000)河北医科大学第二医院消化内科2河北省儿童医院PICU3

背景：肠上皮紧密连接的破坏及其所致的肠黏膜屏障功能受损在肠道疾病的发病中起重要作用,姜黄素对受损的肠黏膜屏障具有保护作用。目的：探讨姜黄素对乙醇诱导的肠黏膜屏障损伤的保护作用。方法：培养Caco-2细胞以制备体外肠上皮细胞屏障模型，将其分为空白对照组、乙醇组和不同浓度姜黄素治疗组(5、20、80 μmol/L)。以单层上皮的跨上皮细胞电阻(TEER)和荧光素钠透过率评估肠上皮屏障功能；蛋白质印迹法和免疫荧光法分别检测Occludin蛋白表达和定位；透射电镜观察细胞结构。结果：与空白对照组相比，乙醇组TEER降低，荧光素钠透过率显著升高(P<0.05)，Occludin蛋白表达显著降低(P<0.05)；免疫荧光法示Occludin蛋白表达不连续，荧光强度减弱；透射电镜示刷状缘排列紊乱，细胞间连接模糊。经姜黄素治疗后，上述指标均显著改善，其中20 μmol/L姜黄素治疗组的疗效最为明显(P<0.05)。结论：姜黄素对乙醇诱导的肠上皮细胞屏障损伤具有保护作用。

关键词姜黄素；肠黏膜屏障；紧密连接；乙醇；治疗

Protective Effect of Curcumin on Intestinal Epithelial Cell Barrier DisruptionWANGGai1,WANGNa2,HAOJingxia3.1PICU,BaodingChildren’sHospital,Baoding,HebeiProvince(071000);2DepartmentofGastroenterology,theSecondHospitalofHebeiMedicalUniversity,Shijiazhuang;3PICU,Children’sHospitalofHebeiProvince,Shijiazhuang

Background: Disruption of intestinal epithelial tight junction and the followed barrier function play important roles in the pathogenesis of intestinal disorders. Curcumin could provide protection for the impaired barrier function. Aims: To investigate the protective effect of curcumin on ethanol-induced intestinal mucosal barrier disruption. Methods: Caco-2 cells were cultured to establish intestinal epithelial cell barrier modelinvitro, and then were divided into control group, ethanol group and different concentrations of curcumin groups (5, 20, 80 μmol/L curcumin). Trans-epithelial electrical resistance (TEER) and flux of sodium fluorescein for Caco-2 cell monolayers were measured to examine intestinal epithelial barrier function. Expression and localization of Occludin protein were measured by Western blotting and immunofluorescence, respectively. Cell structure was observed by transmission electron microscopy (TEM). Results: Compared with control group, TEER was significantly decreased and flux of sodium fluorescein was significantly increased (P<0.05), expression of Occludin protein was significantly decreased (P<0.05) in ethanol group. Immunofluorescence showed that Occludin protein expression was discontinuous and fluorescence intensity was low. TEM showed that brusher border was disorganized, and cell-cell junction was vague. When pretreated with curcumin, the above-mentioned indices were significantly improved, especially in 20 μmol/L curcumin group (P<0.05). Conclusions: Curcumin protects ethanol-induced intestinal epithelial cell barrier disruption.

Key wordsCurcumin;Intestinal Mucosal Barrier;Tight Junctions;Ethanol;Therapy

肠上皮屏障由肠上皮细胞和细胞间紧密连接(tight junction, TJ)构成。Occludin蛋白是肠黏膜TJ蛋白成员之一，其通过胞膜外部分与相邻细胞结合而产生细胞旁封闭，对肠黏膜屏障功能的维持至关重要。各种病理情况均能造成肠黏膜屏障损伤，导致内毒素、细菌入血进而引发全身炎症反应综合征、严重脓毒症、败血症，最终导致多脏器功能衰竭[1]。儿童肠壁薄，通透性高，肠黏膜屏障功能差，故肠黏膜屏障在儿科疾病的发生、发展中意义重大。本研究通过给予乙醇诱导的肠上皮屏障损伤模型不同浓度的姜黄素治疗，旨在探讨姜黄素对肠黏膜屏障的保护作用。……