Expression characteristics of silent information regulator transcript 1 in intestinal tissues of neonatal necrotizing enterocolitis

doi:10.3969/j.issn.1674-8115.2021.09.004

Abstract

Abstract: Objective

·To explore the expression characteristics of silent information regulator transcript 1 (SIRT1) in intestinal tissues of necrotizing enterocolitis (NEC), and preliminarily discuss the effect and possible mechanisms of SIRT1 in NEC.

Methods

·From June 2018 to October 2020, 80 children with NEC who were treated by neonatal surgery in the Children's Hospital, Zhejiang University School of Medicine were divided into observation group and control group. The tissue samples of the observation group were inflammatory necrotic intestinal tubes, while those of the control group were incised intestinal tubes. The NEC children in the control group were with intestinal strictures after conservative treatment, and then were treated by surgery again. The clinical data of the two groups were collected 24 h before surgery, including procalcitonin (PCT), hypersensitive C reactive protein (hs-CRP), cytokines [interleukin-2 (IL-2), IL-4, IL-6, IL-10, tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ)] in the serum, gender, gestational age and so on. The expression characteristics of SIRT1, nuclear factor-κB (NF-κB), transforming growth factor β1 (TGF-β1) and Smad3 proteins of the tissues in the two groups were detected by immunohistochemistry. IEC-6 cells of small intestinal epithelium of SD female rats were cultured in vitro. After SIRT1 expression in IEC-6 cells was inhibited by small interfering RNA (siRNA) technology, the protein expression of NF-κB was detected by Western blotting and the effects of SIRT1 on IEC-6 cells were detected by Cell Counting Kit-8 (CCK8) and Transwell migration assay.

Results

·There was no significant difference in gender, gestational age, birth weight and delivery mode between the two groups. Compared with the control group, the levels of hs-CRP, PCT, IL-6 and IL-10 in the serum in the observation group increased within 24 h before surgery (all P=0.000).Compared with the margin tissues of narrow intestines in the control group, the positive expression of SIRT1 in NEC necrotic intestinal tissues in the observation group showed low expression, and the positive expression of NF-κB was overexpressed (both P=0.000). There was no significant difference in the expression of Smad3 and TGF-β1 between the two groups. The expression of SIRT1 in NEC necrotic intestinal tissue was negatively correlated with the expression of NF-κB (r=-0.592, P=0.000). After inhibiting the mRNA and protein expression of SIRT1, the relative expression of NF-κB in IEC-6 cells was increased, and the proliferation and migration ability of the cells was significantly decreased.

Conclusion

·The mechanism for the SIRT1 reducing progression of NEC may be that SIRT1 can inhibit the expression of NF-κB to reduce the expression of inflammatory factors in NEC, and another possible mechanism may be that SIRT1 can protect intestinal epithelial cells by promoting cell proliferation and migration.

Key words: silencing information regulator factor 1 (SIRT1), nuclear factor-κB (NF-κB), necrotizing enterocolitis (NEC), inflammatory cytokine, neonate

CLC Number:

R722.1

Rui CHEN, Yun ZHAO, Xiao-xia ZHAO, Dong MA, Yi-jiang HAN, Deng-ming LAI, Wei-zhong GU, Jin-fa TOU. Expression characteristics of silent information regulator transcript 1 in intestinal tissues of neonatal necrotizing enterocolitis[J]. JOURNAL OF SHANGHAI JIAOTONG UNIVERSITY (MEDICAL SCIENCE), 2021, 41(9): 1154-1161.

Figures/Tables 7

TrendMD

References 23

1	Niño DF, Sodhi CP, Hackam DJ. Necrotizing enterocolitis: new insights into pathogenesis and mechanisms[J]. Nat Rev Gastroenterol Hepatol, 2016, 13(10): 590-600.
2	Cho SX, Berger PJ, Nold-Petry CA, et al. The immunological landscape in necrotising enterocolitis[J]. Expert Rev Mol Med, 2016, 18: e12.
3	Cho SX, Rudloff I, Lao JC, et al. Characterization of the pathoimmunology of necrotizing enterocolitis reveals novel therapeutic opportunities[J]. Nat Commun, 2020, 11(1): 5794.
4	Agakidou E, Agakidis C, Gika H, et al. Emerging biomarkers for prediction and early diagnosis of necrotizing enterocolitis in the era of metabolomics and proteomics[J]. Front Pediatr, 2020, 8: 602255.
5	Chadha S, Wang LQ, Hancock WW, et al. Sirtuin-1 in immunotherapy: a Janus-headed target[J]. J Leukoc Biol, 2019, 106(2): 337-343.
6	施诚仁, 金先庆, 李仲智. 小儿外科学[M]. 4版. 北京: 人民卫生出版社, 2009: 309-312.
7	张岚, 白铂亮, 王莉, 等. SIRT1信号通路在新生儿坏死性小肠结肠炎肠组织中的表达研究[J]. 海南医学院学报, 2019, 25(8): 578-582.
8	窦敏, 郑英霞, 韩丽, 等. PRMT4在胃癌发生和发展中的作用和机制研究[J]. 上海交通大学学报(医学版), 2020, 40(5): 609-618.
9	Denning NL, Prince JM. Neonatal intestinal dysbiosis in necrotizing enterocolitis[J]. Mol Med, 2018, 24: 4.
10	Hackam DJ, Sodhi CP. Toll-like receptor-mediated intestinal inflammatory imbalance in the pathogenesis of necrotizing enterocolitis[J]. Cell Mol Gastroenterol Hepatol, 2018, 6(2): 229-238.e1.
11	Mihi B, Good M. Impact of Toll-like receptor 4 signaling in necrotizing enterocolitis: the state of the science[J]. Clin Perinatol, 2019, 46(1): 145-157.
12	Lan KC, Chao SC, Wu HY, et al. Salidroside ameliorates sepsis-induced acute lung injury and mortality via downregulating NF‑κB and HMGB1 pathways through the upregulation of SIRT1[J]. Sci Rep, 2017, 7(1): 12026.
13	Li GQ, Xia ZB, Liu Y, et al. SIRT1 inhibits rheumatoid arthritis fibroblast-like synoviocyte aggressiveness and inflammatory response via suppressing NF-κB pathway[J]. Biosci Rep, 2018, 38(3): BSR20180541.
14	Wang L, Wang MM, Dou HJ, et al. Sirtuin 1 inhibits lipopolysaccharide-induced inflammation in chronic myelogenous leukemia k562 cells through interacting with the Toll-like receptor 4-nuclear factor κB-reactive oxygen species signaling axis[J]. Cancer Cell Int, 2020, 20: 73.
15	Mishra M, Duraisamy AJ, Kowluru RA. Sirt1: a guardian of the development of diabetic retinopathy[J]. Diabetes, 2018, 67(4): 745-754.
16	郑文香, 丛立春, 韩斌. 沉默信息调节因子1在具核梭杆菌诱导肠上皮细胞炎症和凋亡中的作用[J]. 中国病原生物学杂志, 2019, 14(6): 643-649, 655.
17	尹海朦, 何鑫, 单颖, 等. 沉默信息调节因子1促进鼻咽癌增殖、迁移和脂质代谢机制的研究[J]. 中华耳鼻咽喉头颈外科杂志, 2020, 55(10): 934-943.
18	Wellman AS, Metukuri MR, Kazgan N, et al. Intestinal epithelial Sirtuin 1 regulates intestinal inflammation during aging in mice by altering the intestinal microbiota[J]. Gastroenterology, 2017, 153(3): 772-786.
19	Markel TA, Martin CA, Chaaban H, et al. New directions in necrotizing enterocolitis with early-stage investigators[J]. Pediatr Res, 2020, 88(): 35-40.
20	Zhang WJ, Zhang YY, Guo XH, et al. Sirt1 protects endothelial cells against LPS-induced barrier dysfunction[J]. Oxid Med Cell Longev, 2017, 2017: 4082102.
21	Hodzic Z, Bolock AM, Good M. The role of mucosal immunity in the pathogenesis of necrotizing enterocolitis[J]. Front Pediatr, 2017, 5: 40.
22	Burge K, Bergner E, Gunasekaran A, et al. The Role of glycosaminoglycans in protection from neonatal necrotizing enterocolitis: a narrative review[J]. Nutrients, 2020, 12(2): 546.
23	Hansen LW, Khader A, Yang WL, et al. Sirtuin 1 activator SRT1720 protects against organ injury induced by intestinal ischemia-reperfusion[J]. Shock, 2016, 45(4): 359-366.

Item	Control group （n=40）	Obeservation group （n=40）	χ²/t value	P value
Gender/n			0.202	0.653
Male	23	21
Female	17	19
Gestational age/n			0.139	0.709
Premature infant（<37 week）	37	35
Term infant（≥37 week）	3	5
Birth weight/g
Premature infant	1 614.1±477.1	1 689.0±493.6	0.109	0.913
Term infant	2 985.0±551.2	3 181.0±583.7	0.224	0.830
Delivery mode/n			0.058	0.809
Vaginal delivery	12	13
Cesarean section	28	27

Item	Control group （n=40）	Obeservation group （n=40）	χ²/t value	P value
Gender/n			0.202	0.653
Male	23	21
Female	17	19
Gestational age/n			0.139	0.709
Premature infant（<37 week）	37	35
Term infant（≥37 week）	3	5
Birth weight/g
Premature infant	1 614.1±477.1	1 689.0±493.6	0.109	0.913
Term infant	2 985.0±551.2	3 181.0±583.7	0.224	0.830
Delivery mode/n			0.058	0.809
Vaginal delivery	12	13
Cesarean section	28	27

Inflammatory factor	Control group （n=40）	Obeservation group （n=40）	U value	P value
hs-CRP/（mg·L^-1）	1.00 （0.50， 2.00）	58.91 （32.74， 107.40）	89.50	0.000
PCT/（ng·mL^-1）	0.25 （0.22， 0.46）	10.25 （6.51， 17.93）	0.00	0.000
IL-2/（pg·mL^-1）	4.70 （2.98， 6.80）	4.55 （3.43， 6.53）	796.00	0.971
IL-4/（pg·mL^-1）	2.15 （1.59， 2.66）	2.29 （1.60， 2.75）	730.00	0.504
IL-6/（pg·mL^-1）	11.85 （9.75， 25.03）	719.30 （156.30， 1 555.00）	20.00	0.000
IL-10/（pg·mL^-1）	3.20 （2.20， 4.95）	23.60 （10.15， 51.08）	0.00	0.000
TNF-α/（pg·mL^－1）	3.09 （2.44， 3.98）	3.27 （2.44， 3.94）	786.00	0.895
IFN-γ/（pg·mL^-1）	3.50 （2.45， 6.93）	5.70 （2.80， 10.27）	614.00	0.074

Inflammatory factor	Control group （n=40）	Obeservation group （n=40）	U value	P value
hs-CRP/（mg·L^-1）	1.00 （0.50， 2.00）	58.91 （32.74， 107.40）	89.50	0.000
PCT/（ng·mL^-1）	0.25 （0.22， 0.46）	10.25 （6.51， 17.93）	0.00	0.000
IL-2/（pg·mL^-1）	4.70 （2.98， 6.80）	4.55 （3.43， 6.53）	796.00	0.971
IL-4/（pg·mL^-1）	2.15 （1.59， 2.66）	2.29 （1.60， 2.75）	730.00	0.504
IL-6/（pg·mL^-1）	11.85 （9.75， 25.03）	719.30 （156.30， 1 555.00）	20.00	0.000
IL-10/（pg·mL^-1）	3.20 （2.20， 4.95）	23.60 （10.15， 51.08）	0.00	0.000
TNF-α/（pg·mL^－1）	3.09 （2.44， 3.98）	3.27 （2.44， 3.94）	786.00	0.895
IFN-γ/（pg·mL^-1）	3.50 （2.45， 6.93）	5.70 （2.80， 10.27）	614.00	0.074

Protein	Control group （n=40）	Observation group （n=40）	χ² value	P value
SIRT1/n			33.885	0.000
+	34	8
-	6	32
NF-κB/n			24.444	0.000
+	11	33
-	29	7
Smad3/n			0.238	0.626
+	27	29
-	13	11
TGF-β1/n			1.067	0.302
+	8	12
-	32	28