黄芪多糖对严重烫伤大鼠肠道免疫功能的影响

[Effects of astragalus polysaccharide on intestinal immune function of rats with severe scald injury].

作者信息

Huang Cuilan, Zhan Jianhua, Luo Jinhua

机构信息

Department of Burns, the First Affiliated Hospital of Nanchang University, Nanchang 330006, China.

Department of Burns, the First Affiliated Hospital of Nanchang University, Nanchang 330006, China. Email:

出版信息

Zhonghua Shao Shang Za Zhi. 2015 Feb;31(1):30-6.

DOI:
Abstract

OBJECTIVE

To observe the effects of astragalus polysaccharide (AP) on the intestinal mucosal morphology, level of secretory IgA (s-IgA) in intestinal mucus, and distribution of T lymphocyte subsets in Peyer's patch in rats with severe scald injury.

METHODS

One hundred and thirty SD rats were divided into sham injury group (SI, sham injured, n = 10), scald group (S, n = 30), low dosage group (LD, n = 30), moderate dosage group (MD, n = 30), and high dosage group (HD, n = 30) according to the random number table. Rats in the latter 4 groups were inflicted with 30% TBSA full-thickness scald on the back. From post injury hour 2, rats in groups LD, MD, and HD were intraperitoneally injected with 0.5 mL AP solution with the dosage of 100, 200, and 300 mg/kg each day respectively, and rats in group S were injected with 0.5 mL normal saline instead. Ten rats from group SI immediately after injury and 10 rats from each of the latter 4 groups on post injury day (PID) 3, 7, 14 were sacrificed, and their intestines were harvested. The morphology of ileal mucosa was examined after HE staining; the level of s-IgA in ileal mucus was determined with double-antibody sandwich ELISA method; the proportions of CD3⁺, CD4⁺, CD8⁺ T lymphocytes in Peyer's patches of intestine were determined with flow cytometer, and the proportion of CD4⁺ to CD8⁺ was calculated. Data were processed with one-way analysis of variance, analysis of variance of factorial design, and SNK test.

RESULTS

(1) Villi in normal form and intact villus epithelial cells were observed in rats of group SI immediately after injury, while edema of villi and necrosis and desquamation of an enormous amount of villi were observed in groups with scalded rats on PID 3, with significant infiltration of inflammatory cells. On PID 7, no obvious improvement in intestinal mucosal lesion was observed in groups with scalded rats. On PID 14, the pathology in intestinal mucosa of rats remained nearly the same in group S, and it was alleviated obviously in groups LD and MD, and the morphology of intestinal mucosa of rats in group HD was recovered to that of group SI. (2) On PID 3, 7, and 14, the level of s-IgA in intestinal mucus significantly decreased in groups S, LD, MD, and HD [(43 ± 5), (45 ± 5), (46 ± 5) µg/mL; (47 ± 5), (48 ± 5), (49 ± 6) µg/mL; (50 ± 6), (51 ± 5), (52 ± 5) µg/mL; (53 ± 6), (54 ± 5), (55 ± 5) µg/mL] as compared with that of rats in group SI immediately after injury [(69 ± 4) µg/mL, with P values below 0.05]. The level of s-IgA in intestinal mucus of rats in group MD was significantly higher than that in group S at each time point (with P values below 0.05), and that of group HD was significantly higher than that in groups S and LD at each time point (with P values below 0.05). (3) Compared with those of rats in group SI immediately after injury, the proportions of CD3⁺ T lymphocytes and CD4⁺ T lymphocytes significantly decreased in groups with scalded rats at each time point (with P values below 0.05), except for those in group HD on PID 14. The proportion of CD4⁺ T lymphocytes of rats in group LD was significantly higher than that in group S on PID 3 (P < 0.05). The proportions of CD3⁺ T lymphocytes and CD4⁺ T lymphocytes were significantly higher in groups MD and HD than in groups S and LD (except for the proportion of CD4⁺ T lymphocytes in group MD on PID 3 and 14) at each time point (with P values below 0.05). The proportion of CD3⁺ T lymphocytes on PID 7 and 14 and that of CD4⁺ T lymphocytes on PID 3 were significantly higher in group HD than in group MD (with P values below 0.05). Compared with that of rats in group SI immediately after injury, the proportion of CD8⁺ T lymphocytes significantly increased in the other 4 groups at each time point (with P values below 0.05). The proportion of CD8⁺ T lymphocytes was significantly lower in rats of group LD on PID 7 and 14 and groups MD and HD at each time point than in group S (with P values below 0.05). The proportion of CD8⁺ T lymphocytes was significantly lower in rats of group MD on PID 7 and 14 and group HD at each time point than in group LD (with P values below 0.05). The proportion of CD8⁺ T lymphocytes was significantly lower in rats of group HD on PID 7 and 14 than in group MD (with P values below 0.05). On PID 3, 7, and 14, the proportion of CD4⁺ to CD8⁺ was significantly lower in groups S, LD, MD, and HD (0.65 ± 0.11, 0.68 ± 0.13, 0.73 ± 0.22; 0.76 ± 0.15, 0.78 ± 0.14, 0.90 ± 0.10; 0.85 ± 0.21, 0.89 ± 0.18, 1.08 ± 0.19; 0.99 ± 0.20, 1.05 ± 0.21, 1.25 ± 0.23) as compared with that of rats in group SI immediately after injury (1.74 ± 0.20, with P values below 0.05). The proportion of CD4⁺ to CD8⁺ was significantly higher in rats of group HD than in group MD on PID 7 (P < 0.05), and the proportion was significantly higher in these two groups than in group S at each time point (with P values below 0.05). The proportion of CD4⁺ to CD8⁺ was significantly higher in rats of group MD on PID 14 and group HD at each time point than in group LD (with P values below 0.05). Compared within each group, the proportions of CD3⁺, CD4⁺, CD8⁺ T lymphocytes and the proportion of CD4⁺ to CD8⁺ of rats in groups LD, MD, and HD showed a trend of gradual elevation along with passage of time.

CONCLUSIONS

AP can improve the injury to intestinal mucosa and modulate the balance of T lymphocyte subsets in Peyer's patch in a time- and dose-dependent manner, and it can promote s-IgA secretion of intestinal mucosa in a dose-dependent manner.

摘要

目的

观察黄芪多糖(AP)对严重烫伤大鼠肠道黏膜形态、肠黏液中分泌型IgA(s-IgA)水平及派尔集合淋巴结中T淋巴细胞亚群分布的影响。

方法

将130只SD大鼠按随机数字表法分为假伤组(SI,假伤,n = 10)、烫伤组(S,n = 30)、低剂量组(LD,n = 30)、中剂量组(MD,n = 30)和高剂量组(HD,n = 30)。后4组大鼠背部给予30%TBSA全层烫伤。自伤后2小时起,LD组、MD组和HD组大鼠分别每天腹腔注射0.5 mL剂量为100、200和300 mg/kg的AP溶液,S组大鼠注射0.5 mL生理盐水。伤后即刻取SI组10只大鼠,伤后第3、7、14天分别取后4组各10只大鼠,处死并摘取肠道。HE染色后观察回肠黏膜形态;采用双抗体夹心ELISA法测定回肠黏液中s-IgA水平;用流式细胞仪检测肠道派尔集合淋巴结中CD3⁺、CD4⁺、CD8⁺ T淋巴细胞比例,并计算CD4⁺/CD8⁺比例。数据采用单因素方差分析、析因设计方差分析及SNK检验进行处理。

结果

(1)伤后即刻SI组大鼠可见形态正常的绒毛及完整的绒毛上皮细胞,烫伤组大鼠伤后第3天可见绒毛水肿,大量绒毛坏死、脱落,有明显炎性细胞浸润。伤后第7天,烫伤组大鼠肠道黏膜损伤无明显改善。伤后第14天,S组大鼠肠道黏膜病理改变基本同前,LD组和MD组明显减轻,HD组大鼠肠道黏膜形态恢复至SI组。(2)伤后第3、7、14天,S组、LD组、MD组和HD组大鼠肠黏液中s-IgA水平均显著低于伤后即刻SI组大鼠[(43±5)、(45±5)、(46±5)μg/mL;(47±5)、(48±5)、(49±6)μg/mL;(50±6)、(51±5)、(52±5)μg/mL;(53±6)、(54±5)、(55±5)μg/mL比(69±4)μg/mL,P值均<0.05]。各时间点MD组大鼠肠黏液中s-IgA水平均显著高于S组(P值<0.05),HD组均显著高于S组和LD组(P值<0.05)。(3)与伤后即刻SI组大鼠相比,各时间点烫伤组大鼠CD3⁺ T淋巴细胞和CD4⁺ T淋巴细胞比例均显著降低(P值<0.05),伤后第14天HD组除外。伤后第3天LD组大鼠CD4⁺ T淋巴细胞比例显著高于S组(P<0.05)。各时间点MD组和HD组CD3⁺ T淋巴细胞和CD4⁺ T淋巴细胞比例均显著高于S组和LD组(伤后第3、14天MD组CD4⁺ T淋巴细胞比例除外,P值<0.05)。伤后第7、14天HD组CD3⁺ T淋巴细胞比例及伤后第3天CD4⁺ T淋巴细胞比例均显著高于MD组(P值<0.05)。与伤后即刻SI组大鼠相比,其他4组各时间点CD8⁺ T淋巴细胞比例均显著升高(P值<0.05)。伤后第7、14天LD组大鼠及各时间点MD组和HD组大鼠CD8⁺ T淋巴细胞比例均显著低于S组(P值<0.05)。伤后第7、14天MD组大鼠及各时间点HD组大鼠CD8⁺ T淋巴细胞比例均显著低于LD组(P值<0.05)。伤后第7、14天HD组大鼠CD8⁺ T淋巴细胞比例显著低于MD组(P值<0.05)。伤后第3、7、14天,S组、LD组、MD组和HD组CD4⁺/CD8⁺比例均显著低于伤后即刻SI组大鼠(0.65±0.11、0.68±0.13、0.73±0.22;0.76±0.15、0.78±0.14、0.90±0.10;0.85±0.21、0.89±0.18、1.08±0.19;0.99±0.20、1.05±0.21、1.25±0.23比1.74±0.20,P值<0.05)。伤后第7天HD组大鼠CD4⁺/CD8⁺比例显著高于MD组(P<0.05),且这两组各时间点均显著高于S组(P值<0.05)。伤后第14天MD组大鼠及各时间点HD组大鼠CD4⁺/CD8⁺比例均显著高于LD组(P值<0.05)。各组内比较,LD组、MD组和HD组大鼠CD3⁺、CD4⁺、CD8⁺ T淋巴细胞比例及CD4⁺/CD8⁺比例均随时间呈逐渐升高趋势。

结论

AP能改善肠道黏膜损伤,呈时间和剂量依赖性调节派尔集合淋巴结中T淋巴细胞亚群平衡,并呈剂量依赖性促进肠黏膜s-IgA分泌。

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