舒芬太尼调节JAK2/STAT3信号通路对烧伤脓毒症大鼠肝损伤的影响

doi:10.11958/20240681

摘要/Abstract

摘要：

目的探讨舒芬太尼（suf）调节Janus激酶2（JAK2）/信号转导和转录激活子3（STAT3）信号通路对烧伤脓毒症大鼠肝损伤的影响。方法将SD大鼠随机分为对照组、模型组、suf低剂量组（0.75 μg/kg）、suf高剂量组（3 μg/kg）、亚胺培南组（90 mg/kg）、suf高剂量+库马霉素组（3 μg/kg suf+4 mg/kg库马霉素）。检测血清肝功能指标丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）水平；HE染色观察肝组织病理变化；酶联免疫吸附试验（ELISA）检测肝组织肿瘤坏死因子-α（TNF-α）、单核细胞趋化蛋白-1（MCP-1）、白细胞介素（IL）-6含量；末端脱氧核苷酸转移酶介导的dUTP尼克末端标记（TUNEL）染色检测肝细胞凋亡情况；Western blot检测肝组织中p-JAK2、p-STAT3蛋白表达。结果与对照组相比，模型组血清ALT、AST水平升高，肝组织中肝细胞肿胀、有大量炎性细胞浸润，肝组织TNF-α、MCP-1、IL-6含量，肝细胞凋亡率及p-JAK2、p-STAT3蛋白表达水平升高（P<0.05）；与模型组相比，suf低、高剂量组和亚胺培南组血清ALT、AST水平降低，肝组织病理损伤有所改善，肝组织TNF-α、MCP-1、IL-6含量，肝细胞凋亡率及p-JAK2、p-STAT3蛋白表达水平降低（P<0.05），且suf高剂量组上述指标改变较suf低剂量组显著（P<0.05），与亚胺培南组差异无统计学意义（P>0.05）；库马霉素抑制了高剂量suf对烧伤脓毒症大鼠肝损伤的改善作用。结论 suf可能通过抑制JAK2/STAT3信号通路改善烧伤脓毒症大鼠肝损伤。

关键词: 烧伤, 脓毒症, 舒芬太尼, Janus激酶2, STAT3转录因子, 炎症, 细胞凋亡

Abstract:

Objective To investigate the effect of sufentanil (suf) on hepatic injury in burn sepsis rats by regulating Janus kinase 2 (JAK2)/signal transduction and transcriptional activator 3 (STAT3) signaling pathway. Methods SD rats were randomly divided into the control group, the model group, the suf low-dose group (0.75 μg/kg), the suf high-dose group (3 μg/kg), the imipenem group (90 mg/kg) and the suf high-dose + kumamycin group (3 μg/kg suf+4 mg/kg kumamycin). Serum liver function indexes including alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected. The pathological changes of liver tissue were observed by HE staining. The contents of tumor necrosis factor-α (TNF-α), monocyte chemoattractant protein-1 (MCP-1) and interleukin (IL)-6 were detected by enzyme-linked immunosorbent assay (ELISA). Terminal deoxynucleotide transferase mediated dUTP nick end labeling (TUNEL) staining was used to detect hepatocyte apoptosis. Western blot assay was used to detect the expression of p-JAK2 and p-STAT3 proteins in liver tissue. Results Compared with the control group, the model group showed elevated serum levels of ALT and AST, swelling of liver cells and infiltration of numerous inflammatory cells in liver tissue. The levels of TNF-α, MCP-1 and IL-6 in liver tissue, apoptosis rate of liver cells, and expression levels of p-JAK2 and p-STAT3 proteins were increased (P<0.05). Compared with the model group, serum levels of ALT and AST were reduced in the suf low-dose group and the suf high-dose group, and pathological damages of liver tissue were improved. The contents of TNF-α, MCP-1 and IL-6 in liver tissue, the apoptosis rate of liver cells, and the expression levels of p-JAK2 and p-STAT3 proteins were reduced (P<0.05). Moreover, changes in above indicators were significantly higher in the suf high dose group of suf than those in the suf low dose group（P<0.05）, and there was no significant difference compared with the imipenem group. Kumamycin inhibited the improvement effect of high-dose suf on liver injury in rats with burn sepsis （P>0.05）. Conclusion Sufentanil may improve liver injury in burn-induced sepsis rats by inhibiting the JAK2/STAT3 signaling pathway.

Key words: burns, sepsis, Sufentanil, Janus kinase 2, STAT3 transcription factor, inflammation, apoptosis

中图分类号:

R631

图/表 6

表1 各组大鼠血清中ALT、AST水平比较（n=12，U/L，$\bar{x} \pm s$）

Tab.1 Comparison of changes in serum ALT and AST levels between six groups of rats

组别	ALT	AST
对照组	66.36±4.58	130.24±8.63
模型组	303.23±15.58^a	412.23±23.34^a
suf低剂量组	266.65±12.36^b	343.66±15.52^b
suf高剂量组	108.83±5.12^bc	175.54±8.33^bc
亚胺培南组	110.22±5.34^bc	180.03±8.55^bc
suf高剂量+库马霉素组	189.95±7.37^d	295.54±12.27^d
F	1 245.098^＊	770.683^＊

图1 大鼠肝组织病理变化的HE染色检测结果（×400）

Fig.1 HE staining results of pathological changes in rat liver tissue (×400)

表2 各组大鼠肝组织中TNF-α、MCP-1和IL-6含量比较（n=6，ng/L，$\bar{x} \pm s$）

Tab.2 Comparison of contents of TNF-α，MCP-1 and IL-6 in liver tissue of rats between six groups

组别	TNF-α	MCP-1	IL-6
对照组	60.23±4.12	82.25±3.79	36.68±1.72
模型组	231.15±10.55^a	245.56±10.11^a	121.22±5.33^a
suf低剂量组	185.57±8.44^b	203.34±9.88^b	102.26±4.75^b
suf高剂量组	106.67±5.03^bc	121.23±5.78^bc	59.96±2.18^bc
亚胺培南组	109.44±5.26^bc	125.56±6.64^bc	61.76±2.95^bc
suf高剂量+ 库马霉素组	165.53±7.21^d	173.59±6.88^d	95.23±4.15^d
F	458.472^＊	384.524^＊	428.566^＊

图2 大鼠肝细胞凋亡变化的TUNEL染色检测结果（×200）

Fig.2 TUNEL staining results of apoptosis changes in rat liver cells (×200)

表3 各组大鼠肝组织中p-JAK2、p-STAT3蛋白表达水平比较（n=6，$\bar{x} \pm s$）

Tab.3 Comparison of p-JAK2 and p-STAT3 protein expression in liver tissues of rats between six groups

组别	p-JAK2/JAK2	p-STAT3/STAT3
对照组	0.36±0.02	0.15±0.01
模型组	0.87±0.08^a	0.73±0.07^a
suf低剂量组	0.69±0.06^b	0.53±0.05^b
suf高剂量组	0.49±0.04^bc	0.28±0.02^bc
亚胺培南组	0.51±0.05^bc	0.29±0.02^bc
suf高剂量+库马霉素组	0.66±0.05^d	0.41±0.04^d
F	68.753^＊	156.279^＊

图3 Western blot检测大鼠肝组织中p-JAK2、p-STAT3蛋白表达 A—F分别为对照组、模型组、suf低剂量组、suf高剂量组、亚胺培南组、suf高剂量+库马霉素组。

Fig.3 Western blot detection results of p-JAK2 and p-STAT3 protein expression in rat liver tissue

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