糖尿病足感染者创面肉芽组织细胞自噬相关蛋白表达

doi:10.3877/cma.j.issn.1674-1358.2018.05.007

目的

分析糖尿病足感染对患者创面肉芽组织细胞自噬相关蛋白表达的影响，为治疗糖尿病足感染提供理论依据。

方法

选取自2013年1月至2017年7月恩施土家族苗族自治州中心医院接诊的88例糖尿病足感染者为研究对象，将正处于感染期44例患者作为观察组，将感染控制后44例患者作为对照组。根据踝肱指数(ABI)分组：观察组患者中20例缺血患者作为缺血组，24例非缺血患者作为非缺血组。提取患者创面组织，进一步处理后采用免疫组织化学染色和蛋白质印迹法进行检测，观察记录患者的姓名、年龄、身高、体重、病程等基本信息，密切观察患者病情变化，详细记录患者创面组织细胞自噬蛋白微管结合轻链蛋白-3(LC3)、自噬特异性基因(Beclin-1)和泛素结合蛋白(p62)等指标。

结果

免疫组织化学染色法显示：观察组患者LC3和Beclin-1水平均显著低于对照组患者(t= 3.638、P = 0.011，t = 2.682、P= 0.022)，p62水平高于对照组患者(t = 4.998、P = 0.009)，差异均有统计学意义。非缺血组患者LC3和Beclin-1水平均显著高于缺血组患者(t= 2.837、P = 0.018，t = 3.028、P = 0.012)，p62水平低于缺血组患者(t = 3.562，P= 0.010)，差异均有统计学意义。观察组患者Beclin-1灰度值低于对照组，差异有统计学意义(t = 3.522，P= 0.013)，而p62灰度值高于对照组，差异有统计学意义(t = 6.927、P= 0.004)；两组患者LC3灰度值差异无统计学意义(t = 1.037、P= 0.056)。

结论

糖尿病足感染会导致患者细胞自噬水平降低，合并缺血患者细胞自噬水平进一步降低，糖尿病足感染控制后，细胞自噬水平升高。

关键词: 糖尿病足感染, 创面肉芽组织, 自噬相关蛋白

Objective

To investigate the influence of diabetic foot infection on the expression of autophagic-related proteins in granulation tissue of wounds, and to provid a theoretical basis for the treatment of diabetic foot infection.

Methods

Total of 88 patients with diabetic foot infection admitted to The Central Hospital of Enshi Tujia and Miao Autonomous Prefecture from January 2013 to July 2017 were selected, among whom, 44 patients in the infection stage were selected as observation group, and 44 patients who were controlled after infection were collected as control group. According to the ankle brachial index (ABI), 44 patients of the observation group were divided into 20 patients with ischemia (ischemic group) and 24 patients with non-ischemia (non-ischemic group). The wound tissue was extracted from these patients. After further treatment, wound tissue were detected by immunohistochemical staining and Western blot, and the patients’name, age, height, weight, course of disease and so on were recorded, respectively, the changes of the patients’condition were observed closely. The levels of autophagy protein microtubule-binding light chain protein-3 (LC3), autophagy specific gene (Beclin-1) and ubiquitin binding protein (p62) in wound tissue were recorded in detail.

Results

Immunohistochemical staining showed that the levels of LC3 and Beclin-1 in the observation group were significantly lower than those of the control group (t= 3.638,P= 0.0112; t = 2.682,P = 0.022), and the level of p62 was higher than that of the control group (t= 4.998,P= 0.009), all with significant differences. The levels of LC3 and Beclin-1 in the non-ischemic group were significantly higher than those of the ischemic group(t= 2.837,P = 0.018; t= 3.028, P= 0.012), and the level of p62 was lower than that of the ischemic group (t= 3.562,P = 0.010), the difference was statistically significant. The gray value of Beclin-1 in the study group was lower than that of the control group (t = 3.522,P= 0.013), while the gray value of p62 in the observation group was higher than that in the control group (t= 3.522, P= 0.013), with significant differences. However, the gray value of p62 was significantly higher than that of the control group (t = 6.927,P = 0.004), but the difference of LC3 gray value between the two groups was not significantly different (t = 1.037, P = 0.056).

Conclusions

The level of autophagy in patients with diabetic foot infection was lower than that in patients with ischemia, and the level of autophagy increased after infection control of diabetic foot.

Key words: Diabetic foot infection, Granulation tissue of wound, Autophagy-related protein

表1 观察组和对照组糖尿病足感染者的一般资料

组别	例数	男/女(例)	年龄(±s，岁)	体质量(±s，kg/m²)	病程(±s，年)
观察组	44	29/15	59.3 ± 13.5	21.6 ± 2.2	3.1 ± 1.3
对照组	44	27/17	58.9 ± 12.9	22.8 ± 2.4	2.9 ± 1.1
统计量	?	χ² = 2.989	t = 2.768	t = 2.243	t = 3.027
P值	?	0.681	0.775	0.597	0.934

表1 观察组和对照组糖尿病足感染者的一般资料

组别	例数	男/女(例)	年龄(±s，岁)	体质量(±s，kg/m²)	病程(±s，年)
观察组	44	29/15	59.3 ± 13.5	21.6 ± 2.2	3.1 ± 1.3
对照组	44	27/17	58.9 ± 12.9	22.8 ± 2.4	2.9 ± 1.1
统计量	?	χ² = 2.989	t = 2.768	t = 2.243	t = 3.027
P值	?	0.681	0.775	0.597	0.934

表2 观察组和对照组患者LC3、Beclin-1和p62免疫组织化学染色(±s)

组别	例数	LC3	Beclin-1	p62
对照组	44	0.88 ± 0.12	0.98 ± 0.16	0.31 ± 0.06
观察组	44	0.44 ± 0.15	0.58 ± 0.13	0.97 ± 0.09
t值	?	3.638	2.682	4.998
P值	?	0.011	0.022	0.009

表2 观察组和对照组患者LC3、Beclin-1和p62免疫组织化学染色(±s)

组别	例数	LC3	Beclin-1	p62
对照组	44	0.88 ± 0.12	0.98 ± 0.16	0.31 ± 0.06
观察组	44	0.44 ± 0.15	0.58 ± 0.13	0.97 ± 0.09
t值	?	3.638	2.682	4.998
P值	?	0.011	0.022	0.009

图2 观察组患者LC3、Beclin-1和p62免疫组织化学染色(× 400)

表3 观察组患者缺血情况对自噬蛋白表达的影响(±s)

患者	例数	LC3	Beclin-1	p62
非缺血组	24	0.96 ± 0.09	1.08 ± 0.12	0.47 ± 0.27
缺血组	20	0.55 ± 0.20	0.66 ± 0.15	0.98 ± 0.07
t值	?	2.837	3.028	3.562
P值	?	0.018	0.012	0.010

表3 观察组患者缺血情况对自噬蛋白表达的影响(±s)

患者	例数	LC3	Beclin-1	p62
非缺血组	24	0.96 ± 0.09	1.08 ± 0.12	0.47 ± 0.27
缺血组	20	0.55 ± 0.20	0.66 ± 0.15	0.98 ± 0.07
t值	?	2.837	3.028	3.562
P值	?	0.018	0.012	0.010

图3 两组患者自噬相关蛋白Western blot分析

表4 观察组和对照组患者自噬相关蛋白Western blot分析灰度值(±s)

组别	例数	LC3	Beclin-1	p62
对照组	44	0.62 ± 0.26	0.59 ± 0.09	0.09 ± 0.04
观察组	44	0.42 ± 0.24	0.37 ± 0.08	0.41 ± 0.16
t值	?	1.037	3.522	6.927
P值	?	0.056	0.013	0.004

表4 观察组和对照组患者自噬相关蛋白Western blot分析灰度值(±s)

组别	例数	LC3	Beclin-1	p62
对照组	44	0.62 ± 0.26	0.59 ± 0.09	0.09 ± 0.04
观察组	44	0.42 ± 0.24	0.37 ± 0.08	0.41 ± 0.16
t值	?	1.037	3.522	6.927
P值	?	0.056	0.013	0.004

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Autophagy-related protein expression in granulation tissue of patients with diabetic foot infection

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Autophagy-related protein expression in granulation tissue of patients with diabetic foot infection