Zhao D X, Ma L, Shen Z A, Li D W, Shang Y R, Yin K, Cheng W F, Wang X, Liu Z X
Burns Institute, the First Hospital Affiliated to the PLA General Hospital, Beijing 100048, China.
Zhonghua Shao Shang Za Zhi. 2016 Dec 20;32(12):752-758. doi: 10.3760/cma.j.issn.1009-2587.2016.12.011.
To observe the secretion function changes of islet beta cells isolated from rats in the early stage of severe scald, and to explore the influence of them. Thirty-six Wistar rats were divided into sham injury (SI) group, sham injury+ exendin-4 (SIE) group, scald (S) group, and scald+ exendin-4 (SE) group according to the random number table, with 9 rats in each group. Rats in groups S and SE were inflicted with 50% total body surface area full-thickness scald by a 12-s immersion of back and a 6-s immersion of abdomen in 94 ℃ hot water. Rats in groups SI and SIE were sham injured through immersion of back and abdomen in 37 ℃ warm water. Rats in groups S and SE were subcutaneously injected with exendin-4 (4 μg/kg) twice a day post injury, while rats in groups SI and SIE were subcutaneously injected with sterile water in the same volume. At post injury hour (PIH) 72, the following detections were performed. Eight rats of each group were respectively selected to measure level of fasting blood glucose with cutting-tail method, and to detect plasma level of glucagon-like peptide 1 (GLP-1) and serum level of insulin by enzyme-linked immunosorbent assay (ELISA). The insulin resistant index (HOMA-IR) was calculated. Six rats of each group were respectively selected for islet isolation. The isolated rat islets were stimulated with RPMI 1640 medium containing 2.8 or 16.7 mmol/L glucose, respectively. Insulin content in supernatant was detected by ELISA, and insulin secretion index was calculated with 6 samples in each group. The isolated islets from 3 rats of each group were selected for the observation of the super-structure of islet beta cells under transmission electron microscope. The number of docked granules in per 10 μm membrane of islet beta cells and the ratio of insulin vesicles to the total insulin granules were calculated with 3 samples in each group. Data were processed with one-way analysis of variance and LSD test. (1) Compared with that in group S, levels of fasting blood glucose of rats in group SI, SIE, and SE were significantly decreased (<0.05 or <0.01). (2) Compared with those in group SI, plasma level of GLP-1 of rats in group SIE was significantly increased (<0.05), while serum level of insulin and HOMA-IR of rats did not change obviously (with values above 0.05). Plasma levels of GLP-1 of rats in groups S and SE were significantly decreased (with values below 0.01), while serum levels of insulin and HOMA-IR were obviously increased (with values below 0.01). Compared with those in group SIE, plasma levels of GLP-1 of rats in groups S and SE were significantly decreased (with values below 0.01), while serum levels of insulin and HOMA-IR were significantly increased (with values below 0.01). Compared with those in group S, plasma level of GLP-1 and serum level of insulin of rats in group SE were significantly increased (with values below 0.01), while HOMA-IR was significantly decreased (<0.05). (3) There was no statistically significant difference in the insulin secretion content of rats in the 4 groups when stimulated with 2.8 mmol/L glucose (>0.05). Under stimulation of 16.7 mmol/L glucose, compared with that in group SI, the insulin secretion content of rats in groups SIE and SE were significantly increased (<0.05 or <0.01), while in group S it was significantly decreased (<0.05). Compared with that in group SE, the insulin secretion content of rats in group S was significantly decreased (<0.01) . Compared with that in group S, the insulin secretion content of rats in group SE was significantly increased (<0.01). Compared with that in group SI (2.25±0.20), the insulin secretion index of rats in group SE (2.68±0.24) was significantly increased (<0.05). Compared with that in group SIE (2.47±0.18), the insulin secretion index of rats in group S (2.11±0.28) was significantly decreased (<0.05). Compared with that in group S, the insulin secretion index of rats in group SE was significantly increased (<0.01). (4) Compared with those in group SI, the number of docked granules per 10 μm membrane of islet beta cells in group SE was significantly increased (<0.05), while the ratio of insulin vesicles of rat islet beta cells in group S was significantly increased (<0.01). Compared with those in group SE, the number of docked granules per 10 μm membrane of islet beta cells in group S was significantly decreased (<0.01), while the ratio of insulin vesicles of rat islet beta cells was significantly increased (<0.05). Compared with those in group S, the number of docked granules per 10 μm membrane of islet beta cells in group SE was significantly increased (<0.01), while the ratio of insulin vesicles of rat islet beta cells was significantly decreased (<0.05). In the early stage of severe scald in rats, level of GLP-1 is decreased and the insulin secretion function of islet beta cells is injured. Long-lasting GLP-1 analogous exendin-4 can improve the secretion function of isolated islet beta cells from severely scalded rats.
观察严重烫伤大鼠早期胰岛β细胞分泌功能变化,并探讨其影响因素。将36只Wistar大鼠按随机数字表法分为假伤(SI)组、假伤+艾塞那肽-4(SIE)组、烫伤(S)组和烫伤+艾塞那肽-4(SE)组,每组9只。S组和SE组大鼠背部浸入94℃热水12秒、腹部浸入6秒造成50%体表面积全层烫伤。SI组和SIE组大鼠背部和腹部浸入37℃温水进行假伤。S组和SE组大鼠伤后每天皮下注射艾塞那肽-4(4μg/kg)2次,SI组和SIE组大鼠皮下注射等量无菌水。伤后72小时进行以下检测。每组分别选取8只大鼠,采用断尾法测定空腹血糖水平,采用酶联免疫吸附测定(ELISA)法检测血浆胰高血糖素样肽1(GLP-1)水平和血清胰岛素水平,并计算胰岛素抵抗指数(HOMA-IR)。每组分别选取6只大鼠进行胰岛分离。将分离的大鼠胰岛分别用含2.8或16.7mmol/L葡萄糖的RPMI 1640培养基刺激。采用ELISA法检测上清液中胰岛素含量,每组6个样本计算胰岛素分泌指数。每组选取3只大鼠分离的胰岛,在透射电镜下观察胰岛β细胞超微结构。每组3个样本计算每10μm胰岛β细胞膜对接颗粒数及胰岛素囊泡与总胰岛素颗粒的比例。数据采用单因素方差分析和LSD检验进行处理。(1)与S组相比,SI组、SIE组和SE组大鼠空腹血糖水平显著降低(<0.05或<0.01)。(2)与SI组相比,SIE组大鼠血浆GLP-1水平显著升高(<0.05),而大鼠血清胰岛素水平和HOMA-IR无明显变化(P值>0.05)。S组和SE组大鼠血浆GLP-1水平显著降低(P值<0.01),而血清胰岛素水平和HOMA-IR显著升高(P值<0.01)。与SIE组相比,S组和SE组大鼠血浆GLP-1水平显著降低(P值<0.01),而血清胰岛素水平和HOMA-IR显著升高(P值<0.01)。与S组相比,SE组大鼠血浆GLP-1水平和血清胰岛素水平显著升高(P值<0.01),而HOMA-IR显著降低(<0.05)。(3)用2.8mmol/L葡萄糖刺激时,4组大鼠胰岛素分泌含量差异无统计学意义(P>0.05)。在16.7mmol/L葡萄糖刺激下,与SI组相比,SIE组和SE组大鼠胰岛素分泌含量显著增加(<0.05或<0.01),而S组显著降低(<0.05)。与SE组相比,S组大鼠胰岛素分泌含量显著降低(<0.01)。与S组相比,SE组大鼠胰岛素分泌含量显著增加(<0.01)。与SI组(2.25±0.20)相比,SE组大鼠胰岛素分泌指数(2.68±0.24)显著增加(<0.05)。与SIE组(2.47±0.18)相比,S组大鼠胰岛素分泌指数(2.11±0.28)显著降低(<0.05)。与S组相比,SE组大鼠胰岛素分泌指数显著增加(<0.01)。(4)与SI组相比,SE组每10μm胰岛β细胞膜对接颗粒数显著增加(<0.05),而S组大鼠胰岛β细胞胰岛素囊泡比例显著增加(<0.0