He Dan, Qiu Minshan, Wang Shan, Yin Haiyan
Department of Anesthesiology, Hengyang Women and Children's Hospital, Hengyang 421001, Hunan, China.
Department of Critical Care Medicine, the First People's Hospital of Dongguan, Dongguan 523000, Guangdong, China.
Zhonghua Wei Zhong Bing Ji Jiu Yi Xue. 2021 Sep;33(9):1052-1056. doi: 10.3760/cma.j.cn121430-20210105-00008.
To study the effect of curcumin on enterocyte apoptosis and its protective effect on intestinal mucosal barrier in septic rats.
Eighty-seven 3-month male Sprague-Dawley (SD) rats were divided into Sham group, model group and curcumin group by random number table method, with 29 rats in each group. The septic rat model was reproduced by cecal ligation and puncture (CLP). 4 mL dimethyl sulfoxide solution were intraperitoneally injected in both Sham group and model group, 200 mg/kg curcumin dissolved by 4 mL dimethyl sulfoxide solution were intraperitoneally injected in curcumin group 10 minutes after operation. The blood samples (15 rats in each group) were collected 2, 12, 24 hours after operation, and the levels of serum procalcitonin (PCT), tumor necrosis factor-α (TNF-α), D-lactic acid and diamine oxidas (DAO) were tested by enzyme linked immunosorbent assay (ELISA). The ileum tissues were collected 12 hours, 24 hours after operation in three groups, water content was tested by weighting, pathologic structure was observed by light microscope, the enterocyte apoptosis was tested by terminal deoxynucleotidyl transferase-mediated dUTP-nick end labeling method (TUNEL). The 7-day survival rate was observed in three groups (14 rats in each group).
The serum levels of PCT, TNF-α, D-lactic acid and DAO were higher in model group at 2, 12, 24 hours after operation than those in Sham group, PCT, TNF-α levels were significantly higher in model group than those in Sham group 2 hours after operation [PCT (μg/L): 1.89±0.17 vs. 0.10±0.02, TNF-α (ng/L): 216.51±1.47 vs. 85.25±8.20, both P < 0.01], D-lactic acid, DAO levels were significantly higher in model group than those in Sham group 12 hours after operation [D-lactic acid (mg/L): 40.53±7.76 vs. 11.29±1.28, DAO (ng/L): 1 120.40±302.35 vs. 330.02±81.28, both P < 0.01]. Compared with model group, the levels of serum PCT, TNF-α, D-lactic acid and DAO were lower in curcumin group 2, 12, 24 hours after operation, the statistical difference appeared from 12 hours after operation [PCT (μg/L): 5.37±0.44 vs. 8.67±0.64, TNF-α (ng/L): 211.12±4.31 vs. 313.30±18.46, D-lactic acid (mg/L): 29.74±1.41 vs. 40.53±7.76, DAO (ng/L): 810.71±201.41 vs. 1 120.40±302.35, all P < 0.05], curcumin group had lower water content in ileum tissues 12 hours, 24 hours after operation [(68.34±0.68)% vs. (70.55±0.87)%, (69.41±0.59)% vs. (71.69±0.87)%, both P < 0.05]. The pathologic structures of intestinal villus were normal in Sham group, however, in model group intestinal villus were atrophic, edematous and shorten 12 hours after operation, it was further exacerbated 24 hours after operation. Compared with model group, the pathologic structures of intestinal villus in curcumin group were relived 12 hours, 24 hours after operation. The number of apoptotic enterocytes were significantly increased in model group compared with Sham group 24 hours after operation (cells: 25.48±6.10 vs. 4.00±2.04, P < 0.05), and the number of apoptotic enterocytes was lower in curcumin group than that in model group at the same time (cells: 15.48±3.75 vs. 25.48±6.10), the difference was statistically significant (both P < 0.05). Seven-day survival rate was significantly lower in curcumin than that in model group [42.9% (6/14) vs. 50.0% (7/14)], however, the difference was not statistically significant (P > 0.05).
Curcumin can protect the intestinal mucosal barrier by inhibiting enterocyte apoptosis in septic rats.
研究姜黄素对脓毒症大鼠肠上皮细胞凋亡的影响及其对肠黏膜屏障的保护作用。
采用随机数字表法将87只3月龄雄性Sprague-Dawley（SD）大鼠分为假手术组、模型组和姜黄素组，每组29只。采用盲肠结扎穿孔（CLP）法复制脓毒症大鼠模型。假手术组和模型组腹腔注射4 mL二甲基亚砜溶液，姜黄素组于术后10分钟腹腔注射用4 mL二甲基亚砜溶液溶解的200 mg/kg姜黄素。术后2、12、24小时采集血样（每组15只大鼠），采用酶联免疫吸附测定（ELISA）法检测血清降钙素原（PCT）、肿瘤坏死因子-α（TNF-α）、D-乳酸和二胺氧化酶（DAO）水平。三组分别于术后12、24小时采集回肠组织，称重法检测含水量，光镜观察病理结构，采用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法（TUNEL）检测肠上皮细胞凋亡情况。观察三组（每组14只大鼠）的7天生存率。
术后2、12、24小时模型组血清PCT、TNF-α、D-乳酸和DAO水平均高于假手术组，术后2小时模型组PCT、TNF-α水平显著高于假手术组[PCT（μg/L）：1.89±0.17 vs. 0.10±0.02，TNF-α（ng/L）：216.51±1.47 vs. 85.25±8.20，均P<0.01]，术后12小时模型组D-乳酸、DAO水平显著高于假手术组[D-乳酸（mg/L）：40.53±7.76 vs. 11.29±1.28，DAO（ng/L）：1 120.40±302.35 vs. 330.02±81.28，均P<0.01]。与模型组相比，姜黄素组术后2、12、24小时血清PCT、TNF-α、D-乳酸和DAO水平均较低，术后12小时出现统计学差异[PCT（μg/L）：5.37±0.44 vs. 8.67±0.64，TNF-α（ng/L）：211.12±4.31 vs. 313.30±18.46，D-乳酸（mg/L）：29.74±1.41 vs. 40.53±7.76，DAO（ng/L）：810.71±201.41 vs. 1 120.40±302.35，均P<0.05]，姜黄素组术后12、24小时回肠组织含水量较低[（68.34±0.68）% vs. （70.55±0.87）%，（69.41±0.59）% vs. （71.69±0.87）%，均P<0.05]。假手术组肠绒毛病理结构正常，而模型组术后12小时肠绒毛萎缩、水肿、缩短，术后24小时进一步加重。与模型组相比，姜黄素组术后12、24小时肠绒毛病理结构有所缓解。术后24小时模型组凋亡肠上皮细胞数量显著多于假手术组（细胞数：25.48±6.10 vs. 4.00±2.04，P<0.05），同一时间姜黄素组凋亡肠上皮细胞数量低于模型组（细胞数：15.48±3.75 vs. 25.48±6.10），差异有统计学意义（均P<0.05）。姜黄素组7天生存率低于模型组[42.9%（6/14） vs. 50.0%（7/14）]，但差异无统计学意义（P>0.05）。
姜黄素可通过抑制脓毒症大鼠肠上皮细胞凋亡保护肠黏膜屏障。
