NF-κB/JNK/PI3K信号表达在糖尿病足多重耐药金黄色葡萄球菌感染中的作用

Role of NF-κB/JNK/PI3K signaling pathway related molecules in multi-drug resistant Staphylococcus aureus infection in diabetic foot

目的研究核因子-κB(NF-κB)/c-Jun N-末端激酶(JNK)/磷酸酰肌醇3-激酶(PI3K)信号表达在糖尿病足多重耐药金黄色葡萄球菌(SA)感染中的作用。方法前瞻性选取2019年7月-2021年7月本院收治的69例SA感染的糖尿病足患者作为研究组,同期选取于本院就诊的40例足部烧伤患者作为对照组。对患者进行SA分离鉴定并做药敏试验,根据耐药结果将研究组分为多重耐药组与非多重耐药组;采用蛋白质印迹法(Western blot)检测NF-κB p65、JNK、PI3K信号通路分子表达强度,使用实时定量PCR检测其mRNA表达量。结果对照组与研究组中性粒细胞比率、血肌酐、C反应蛋白、降钙素原、白介素-6水平差异无统计学意义(P>0.05);研究组糖化血红蛋白、白细胞计数、红细胞沉降率高于对照组(P<0.05)。药敏试验显示,研究组检出33例多重耐药SA,对照组检出2例,研究组多重耐药SA检出率高于对照组(P<0.05)。多重耐药组白细胞计数、中性粒细胞比率、C反应蛋白、降钙素原、白介素-6水平高于非多重耐药组(P<0.05);两组的糖化血红蛋白、红细胞沉降率、血肌酐水平差异无统计学意义(P>0.05)。对照组标本中NF-κB、JNK、PI3K蛋白表达微弱,研究组标本中NF-κB、JNK、PI3K蛋白表达强烈;多重耐药组标本中的NF-κB、JNK、PI3K光密度值高于非多重耐药组。研究组NF-κB、JNK、PI3K表达量高于对照组(P<0.05),且多重耐药组表达量高于非多重耐药组(P<0.05)。结论SA感染的糖尿病足患者创面肉芽组织中NF-κB/JNK/PI3K信号表达增强,且多重耐药SA感染患者增强更显著;SA多重耐药发生可能与炎症反应增强有关。

Objective To investigate the role of nuclear factor-kappa B(NF-κB)/c-Jun N-terminal kinase(JNK)/phosphoinositide 3-kinase(PI3K)signaling pathway related molecules in multi-drug resistant Staphylococcus aureus(SA)infection in diabetic foot.Methods Sixty-nine SA-infected diabetic foot patients admitted to our hospital from July 2019 to July 2021 were prospectively selected as study group,and another 40 patients with foot burns were selected as control group.SA strains were isolated and identified from the patients and drug sensitivity test was conducted.According to the drug resistance results,study group was divided into multiple drug resistance group and non-multiple drug resistance group.The protein expression intensity and mRNA expression levels of NF-κB P65,JNK and PI3K signaling pathway molecules were detected by Western blot and real-time quantitative PCR.Results The neutrophil-lymphocyte ratio,serum creatinine,C-reactive protein,procalcitonin and interleukin-6 levels demonstrated no significant difference between the two groups(P>0.05).The glycated haemoglobin,white blood cell count and erythrocyte sedimentation rate were(5.34±0.61)%,(7.86±1.49)10~9/Land(34.48±7.59)mm/h in control group,which had significant difference with(7.89±1.17)%,(9.42±1.66)10~9/Land(37.52±9.86)mm/h in study group(P<0.05).Drug sensitivity test showed that 33 cases of multidrug-resistant SA infections were detected in study group and 2 cases in control group,and the overall detection rate of multidrug-resistant SA infection was 47.83%in study group,which was higher than 5.00%in control group(P<0.05).The white blood cell count,neutrophil-lymphocyte ratio and C-reactive protein,procalcitonin and interleukin-6 level in multi-drug resistance group were higher than those in non-multi-drug resistance group(P<0.05),while the glycated haemoglobin,erythrocyte sedimentation rate and serum creatinine yielded no statistical difference between the two groups(P>0.05).The expression of NF-κB,JNK,PI3K protein was weak in control group,while was relatively strong in study group.The optical density values of NF-κB,JNK and PI3K in multi-drug resistant group were higher than those in non-multi-drug resistant group.The expression levels of NF-κB,JNK and PI3K were(137.48±16.71),(2.04±0.27)and(2.17±0.29)in study group,which were significantly higher than(93.52±11.36),(0.89±0.06)and(1.38±0.20)in control group(P<0.05).The expression levels of NF-κB,JNK and PI3K were(139.52±17.12),(2.21±0.35)and(2.36±0.41)in multi-drug resistant group,which were significantly higher than(128.37±14.08),(1.76±0.18)and(1.84±0.26)in non-multi-drug resistant group(P<0.05).Conclusion NF-κB/JNK/PI3K signaling expression in granulation tissue of SA infected diabetic foot is enhanced,especially in patients with multidrug resistant SA infection,moreover,the occurrence of multidrug resistance SA infection may be related to the enhancement of inflammatory response.