Distribution of pathogenic bacteria in incisional infections of closed lower limb fracture and expression analysis of miR-15b-5p/IKBKB signaling pathway-related molecules

doi:10.11958/20252915

Abstract

Abstract:

Objective To investigate the distribution of pathogenic bacteria in postoperative incision infection following closed lower limb fracture and the effects of different pathogens on the expression of microRNA-15b-5p (miR-15b-5p)/inhibitor of nuclear factor kappa-B kinase subunit beta (IKBKB) signaling pathway-related molecules. Methods A total of 2 318 patients with closed lower limb fracture were analyzed for postoperative incision infection, with 120 patients diagnosed with incision infection. These patients were divided into the methicillin-resistant Staphylococcus aureus (MRSA) group (group A), the methicillin-sensitive S. aureus (MSSA) group (group B), the coagulase-negative staphylococci (CNS) group (group C) and the other bacterial groups (group D) based on the pathogens. The sensitivity of these pathogens to commonly used antibiotics in each group was analyzed. Infected incision tissue specimens were collected, and the expression levels of miR-15b-5p, IKBKB mRNA and protein were detected using qRT-PCR and Western blot assay to analyze the expression differences between the groups. Results The incidence of incision infection in this group was 5.18%. Among the 120 infected patients, Staphylococcus aureus infection was identified in 84 cases (70.00%). There were no statistically significant differences in the sensitivity of the pathogens to cefazolin sodium and cefuroxime sodium between the groups (P > 0.05). There were significant differences in the sensitivity differences of pathogens to cefazolin sodium, cefuroxime sodium and cefuroxime sodium between different groups (P < 0.01). Compared to the group A, the differences in the sensitivity to cefazolin sodium and cefuroxime sodium were statistically significant in the groups B, C and D (P < 0.01). Compared to the group B, the differences in the sensitivity to cefazolin sodium and cefuroxime sodium were also statistically significant in the groups C and D (P < 0.01). The expression levels of miR-15b-5p were lower in the group C and the group D than those of the group A and the group B (P < 0.01). The expression levels of IKBKB mRNA and protein were higher in the group C and the group D than those in the group A and the group B (P < 0.01). Conclusion Staphylococcus aureus is the most common bacterium causing postoperative incision infections following closed lower limb fracture. It can induce the increased expression of miR-15b-5p in tissue and may promote inflammatory progression by regulating the expression of IKBKB.

Key words: fractures, closed, staphylococcus aureus, microbial sensitivity tests, microRNAs, I-kappa B proteins, surgical site infection

CLC Number:

Figures/Tables 5

References 20

[1]	WANG X, ZHANG M, ZHU T, et al. Flourishing antibacterial strategies for osteomyelitis therapy[J]. Adv Sci (Weinh), 2023, 10(11):e2206154. doi:10.1002/advs.202206154.
[2]	POLMEAR M, HAGEN J E, SCHRANK G M, et al. Deep infections after open and closed fractures[J]. J Bone Joint Surg Am, 2025, 107(Suppl 1):71-79. doi:10.2106/JBJS.24.01249.
[3]	WANG Y, LI H, JIANG S, et al. The glycolytic enzyme PFKFB3 drives kidney fibrosis through promoting histone lactylation-mediated NF-κB family activation[J]. Kidney Int, 2024, 106(2):226-240. doi:10.1016/j.kint.2024.04.016.
[4]	FISCHER A, KRAMER M M, BAÑOS M, et al. Activation of NF-κB signaling by optogenetic clustering of IKKα and β[J]. Adv Biol (Weinh), 2025, 9(9):e00384. doi:10.1002/adbi.202400384.
[5]	JOHANSSON K, GAGNON J D, ZHOU S K, et al. An essential role for miR-15/16 in Treg suppression and restriction of proliferation[J]. Cell Rep, 2023, 42(10):113298. doi:10.1016/j.celrep.2023.113298.
[6]	MARTINO E, D'ONOFRIO N, BALESTRIERI A, et al. MiR-15b-5p and PCSK9 inhibition reduces lipopolysaccharide-induced endothelial dysfunction by targeting SIRT4[J]. Cell Mol Biol Lett, 2023, 28:66. doi:10.1186/s11658-023-00482-5.
[7]	Major Extremity Trauma Research Consortium (METRC). Effect of supplemental perioperative oxygen on SSI among adults with lower-extremity fractures at increased risk for infection:a randomized clinical trial[J]. J Bone Joint Surg Am, 2022, 104(14):1236-1243. doi:10.2106/JBJS.21.01317.
[8]	SLOBOGEAN G P, SPRAGUE S, WELLS J, et al. Effectiveness of iodophor vs chlorhexidine solutions for surgical site infections and unplanned reoperations for patients who underwent fracture repair:the PREP-IT master protocol[J]. JAMA Netw Open, 2020, 3(4):e202215. doi:10.1001/jamanetworkopen.2020.2215.
[9]	COSTA M L, ACHTEN J, KNIGHT R, et al. Effect of incisional negative pressure wound therapy vs standard wound dressing on deep surgical site infection after surgery for lower limb fractures associated with major trauma:the WHIST randomized clinical trial[J]. JAMA, 2020, 323(6):519-526. doi:10.1001/jama.2020.0059.
[10]	SHEN J, SUN D, FU J, et al. Management of surgical site infection post-open reduction and internal fixation for tibial plateau fractures[J]. Bone Joint Res, 2021, 10(7):380-387. doi:10.1302/2046-3758.107.BJR-2020-0175.R2.
[11]	吴晖, 方真华, 潘娜, 等. 开放性与闭合性下肢骨折术后切口感染及血清炎症因子变化[J]. 中华医院感染学杂志, 2020, 30(5):721-724.
	WU H, FANG Z H, PAN N, et al. Study of postoperative incision infection and variation levels of serum inflammatory factor in patient with open and closed lower limb fractures[J]. Chin J nosocomiol, 2020, 30(5):721-724. doi:10.11816/cn.ni.2020-183310.
[12]	MEHDI M, MEHDI T, KIANA S, et al. Prevalence of Bacterial surgical site infection in traumatic patients undergoing orthopedic surgeries:a cross-sectional study[J]. Int J Burns Trauma, 2021, 11(3):191-196.
[13]	MELLINGHOFF S C, BRUNS C, ALBERTSMEIER M, et al. Staphylococcus aureus surgical site infection rates in 5 European countries[J]. Antimicrob Resist Infect Control, 2023, 12(1):104. doi:10.1186/s13756-023-01309-w.
[14]	吴昊, 石展英, 李兵, 等. 头孢呋辛与他类抗菌药物在骨科手术感染患者中的应用效果与效价评估[J]. 中华医院感染学杂志, 2017, 27(23):5399-5402,5410.
	WU H, SHI Z Y, LI B, et al. Application effect observation and titer evaluation of cefuroxime and other antimicrobial agents in patients with orthopedic surgery[J]. Chin J nosocomiol, 2017, 27(23):5399-5402,5410. doi:10.11816/cn.ni.2017-171045.
[15]	邝俊健, 黄献京, 叶义平, 等. 骨科择期清洁切口手术3种头孢菌素预防性应用临床随机研究[J]. 中国医院药学杂志, 2017, 37(2):172-176.
	KUANG J J, HUANG X J, YE Y P, et al. Antibiotic prophylaxis of three cephalosporins in elective clean incision orthopedic surgery: a randomized clinical study[J]. Chin Hosp Pharm J, 2017, 37(2):172-176. doi:10.13286/j.cnki.chinhosppharmacyj.2017.02.17.
[16]	POLADIAN N, ORUJYAN D, NARINYAN W, et al. Role of NF-κB during Mycobacterium tuberculosis infection[J]. Int J Mol Sci, 2023, 24(2):1772. doi:10.3390/ijms24021772.
[17]	ZHANG Z, GUAN X, SUN L. A novel teleost microRNA regulates autophagy and NF-κB activation during bacterial infection[J]. Fish Shellfish Immunol, 2023, 137:108778. doi:10.1016/j.fsi.2023.108778.
[18]	CAO Y, TANG S, NIE X, et al. Decreased miR-214-3p activates NF-κB pathway and aggravates osteoarthritis progression[J]. EBioMedicine, 2021, 65:103283. doi:10.1016/j.ebiom.2021.103283.
[19]	GONZÁLEZ-LÓPEZ P, ÁLVAREZ-VILLARREAL M, RUIZ-SIMÓN R, et al. Role of miR-15a-5p and miR-199a-3p in the inflammatory pathway regulated by NF-κB in experimental and human atherosclerosis[J]. Clin Transl Med, 2023, 13(8):e1363. doi:10.1002/ctm2.1363.
[20]	RAMIREZ H A, PASTAR I, JOZIC I, et al. Staphylococcus aureus triggers induction of miR-15B-5P to diminish DNA repair and deregulate inflammatory response in diabetic foot ulcers[J]. J Invest Dermatol, 2018, 138(5):1187-1196. doi:10.1016/j.jid.2017.11.038.

组别	n	性别（男/女）	年龄/ 岁	手术部位/例
组别	n	性别（男/女）	年龄/ 岁	股骨	胫腓骨	踝关节	足部
A组	44	25/19	55.05±12.05	2	20	7	15
B组	40	23/17	55.73±11.75	0	22	10	8
C组	17	12/5	52.65±13.67	0	8	6	3
D组	19	11/8	53.32±15.33	0	9	5	5
χ²或F		1.077	0.318	0.625^▲

组别	n	性别（男/女）	年龄/ 岁	手术部位/例
组别	n	性别（男/女）	年龄/ 岁	股骨	胫腓骨	踝关节	足部
A组	44	25/19	55.05±12.05	2	20	7	15
B组	40	23/17	55.73±11.75	0	22	10	8
C组	17	12/5	52.65±13.67	0	8	6	3
D组	19	11/8	53.32±15.33	0	9	5	5
χ²或F		1.077	0.318	0.625^▲

基因	引物序列（5'→3'）	产物大小/bp
IKBKB	上游：GGGAAATAACACCCGAAAG 下游：TGTAACTGAACATAAAC	165
miR-15b-5p	上游：AAAGCAAATCATCGGACGACC 下游：GTACAACACATTGTTTCCTCGGA	181
GAPDH	上游：TGCACCACCAACTGCTTAGC 下游：GGCATGGACTGTGGTCATGAG	87

基因	引物序列（5'→3'）	产物大小/bp
IKBKB	上游：GGGAAATAACACCCGAAAG 下游：TGTAACTGAACATAAAC	165
miR-15b-5p	上游：AAAGCAAATCATCGGACGACC 下游：GTACAACACATTGTTTCCTCGGA	181
GAPDH	上游：TGCACCACCAACTGCTTAGC 下游：GGCATGGACTGTGGTCATGAG	87

组别	n	头孢唑林钠		头孢呋辛钠		P
组别	n	敏感	耐药	敏感	耐药	P
A组	44	0	44	0	44	1.000^▲
B组	40	37	3	35	5	0.712^▲
C组	17	4	13	3	14	1.000^▲
D组	19	5	14	9	10	0.313^▲
χ²		79.736^＊＊		71.388^＊＊