| 摘要: |
| 目的：探讨五味子乙素（Sch B） 调控miR-22-3p/runt 相关转录因子3（RUNX3） 分子轴对重症急性胰腺炎（SAP）腺泡细胞损伤的影响。方法：将大鼠胰腺腺泡细胞AR42J 分为Con 组、SAP 组、SAP+Sch-L 组、SAP+Sch-M 组、SAP+Sch-H组、SAP+anti-miR-NC 组、SAP+anti-miR-22-3p 组、SAP+Sch+miR-NC 组、SAP+Sch+miR-22-3p 组。除Con 组外，其余各组采用10 nmol/L 雨蛙素联合10 mg/L 脂多糖刺激AR42J 细胞24 h 构建SAP 细胞模型。SAP+Sch-L 组、SAP+Sch-M 组、SAP+Sch-H 组采用浓度为10、20、40 μmol/L 的Sch B 处理细胞；SAP+anti-miR-NC 组、SAP+anti-miR-22-3p 组为分别转染antimiR-NC、anti-miR-22-3p；SAP+Sch+miR-NC 组、SAP+Sch+miR-22-3p 组为分别转染miR-NC、miR-22-3p mimics 并用40 μmol/L的Sch B 干预。酶联免疫吸附法测定细胞培养液上清中白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α） 表达水平；流式细胞术检测细胞凋亡率；实时荧光定量PCR 检测miR-22-3p 和RUNX3 mRNA 表达水平；BCA 法检测RUNX3、B 细胞淋巴瘤-2 基因（Bcl-2）、Bcl-2 相关X 蛋白（Bax）、活化的含半胱氨酸的天冬氨酸蛋白水解酶3（cleaved-caspase3） 表达水平；双荧光素酶报告基因实验检测miR-22-3p 与RUNX3 的靶向结合关系。结果：与Con 组比较，SAP 组AR42J 细胞IL-6、TNF-α、miR-22-3p、Bax、cleaved-caspase3 的表达及细胞凋亡率升高（P＜0.05），RUNX3、Bcl-2 的表达降低（P＜0.05）。与SAP 组比较，SAP+Sch-L组、SAP+Sch-M 组、SAP+Sch-H 组IL-L、TNF-α、miR-22-3P、Bcl-2 表达降低（P＜0.05），Bax、cleaved-caspase3、RUNX3 表达及细胞凋亡率升高（P＜0.05），且呈剂量依赖性（P＜0.05）。与SAP+anti-miR-NC 组比较，SAP+anti-miR-22-3p 组细胞IL-6、TNF-α、Bcl-2 的表达降低（P＜0.05），细胞凋亡率及Bax、cleaved-caspase3、RUNX3 的表达升高（P＜0.05）。与SAP+Sch+miR-NC 组比较，SAP+Sch+miR-22-3p 组细胞IL-6、TNF-α、Bcl-2 的表达升高（P＜0.05），细胞凋亡率及Bax、cleavedcaspase3、RUNX3 的表达降低（P＜0.05）。结论：Sch B 可通过下调miR-22-3p/RUNX3 分子轴减轻SAP 腺泡细胞损伤。 |
| 关键词: 重症急性胰腺炎 五味子乙素 miR-22-3p RUNX3 细胞凋亡 细胞实验 |
| DOI： |
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| 基金项目: |
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| Study on Effect of Schizandrin B Regulating MiR- 22- 3p/RUNX3 Molecular Axis onInjury of Acinar Cells with Severe Acute Pancreatitis |
| YIN Luxi,DING Wen,WANG Xiaohua |
| (单篇PDF全文查阅请上中国知网下载阅读（中国知网电子版刊出时间：纸刊刊出日期一个月左右）) |
| Abstract: |
| Abstract： Objective： To discuss the effect of Schizandrin B(Sch B) regulating miR- 22- 3p/runt- related transcription
factor 3(RUNX3) molecular axis on injury of acinar cells with severe acute pancreatitis(SAP). Methods： The pancreatic
acinar cells AR42J of rats were divided into the Con group，the SAP group，the SAP + Sch- L group，the SAP + Sch-M
group，the SAP+Sch-H group，the SAP+anti-miR-NC group，the SAP+anti-miR-22-3p group，the SAP+Sch+miR-NC
group，and the SAP+Sch+miR-22-3p group. Except for the Con group，the other groups were given caerulein at a dose
of 10 nmol/L and lipopolysaccharide at a dose of 10 mg/L to stimulate AR42J cells for 24 hours to establish SAP cell
model. In the SAP+Sch-L group，the SAP+Sch-M group，and the SAP+Sch-H group，the cells were treated with Sch B
at concentrations of 10，20，and 40 μmol/L; in the SAP+anti-miR-NC group and the SAP+anti-miR-22-3p group，the
cells were transfected with anti-miR-NC and anti-miR-22-3p respectively; in the SAP+Sch+miR-NC group and the SAP+
Sch+miR-22-3p group，the cells were transfected with miR-NC and miR-22-3p mimics respectively and intervened with
Sch B at a concentration of 40 μmol/L. The expression levels of interleukin- 6(IL-6) and tumor necrosis factor α(TNF-α) in
the culture supernatant of cells were detected by enzyme linked immunosorbent assay. The rate of cell apoptosis was
detected by flow cytometry. The mRNA expression levels of miR- 22- 3p and RUNX3 were detected by real- time
fluorescence quantitative PCR. The expression levels of RUNX3，B- cell lymphoma- 2(Bcl- 2)，Bcl- 2- associated X protein
(Bax)，and cleaved-cysteinyl aspartate specific proteinase 3(cleaved-caspase3) were detected by BCA method. The targeted
binding relationship between miR- 22- 3p and RUNX3 was detected by dual- luciferase reporter assay. Results：Compared
with those in the Con group，in the SAP group，the expression levels of IL-6，TNF-α，miR-22-3p，Bax，and cleavedcaspase3
in AR42J cells as well as the rate of cell apoptosis were increased(P＜0.05)，and the expression levels of RUNX3
and Bcl- 2 were decreased(P＜0.05). Compared with those in the SAP group， in the SAP + Sch- L group， SAP + Sch- M
group and SAP+Sch-H group，the expression levels of IL-6，TNF- α，miR-22-3p and Bcl-2 were decreased(P＜0.05)，
and the expression levels of Bax，cleaved- caspase3，and RUNX3 as well as the rate of cell apoptosis were increased(P＜
0.05)，and in a dose-dependent manner(P＜0.05). Compared with those in the SAP+anti-miR-NC group，in the SAP+antimiR-
22-3p group，the expression levels of IL- 6，TNF-α，and Bcl-2 were decreased(P＜0.05)；the expression levels of
Bax，cleaved-caspase3，and RUNX3 as well as the rate of cell apoptosis were increased(P＜0.05). Compared with those in
the SAP+Sch+miR-NC group，in the SAP+Sch+miR-22-3p group，the expression levels of IL-6，TNF-α，and Bcl-2 were
increased(P＜0.05)；the expression levels of Bax，cleaved-caspase3，and RUNX3 as well as the rate of cell apoptosis were
decreased(P＜0.05). Conclusion：Sch B can relieve the injury of acinar cells with SAP by down-regulating miR-22-3p/RUNX3
molecular axis. |
| Key words: Keywords：Severe acute pancreatitis Schizandrin B MiR-22-3p RUNX3 Cell apoptosis Cell experiment |
