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中华细胞与干细胞杂志(电子版) ›› 2022, Vol. 12 ›› Issue (01) : 8 -13. doi: 10.3877/cma.j.issn.2095-1221.2022.01.002

论著

PSP下调miR-345-5p对雨蛙素诱导的AP腺泡细胞氧化应激和炎症因子表达
倪昊1,(), 王发武2, 李翔天2, 马战胜1   
  1. 1. 210046 南京泰康仙林鼓楼医院急诊科
    2. 211800 南京市浦口区中医院普外科
  • 收稿日期:2021-03-15 出版日期:2022-02-01
  • 通信作者: 倪昊

Polygonatum sibiricum polysaccharide inhibits caerulein-induced oxidative stress and inflammatory factors expression of acinar cells in acute pancreatitis by down-regulating miR-345-5p

Hao Ni1,(), Fawu Wang2, Xiangtian Li2, Zhansheng Ma1   

  1. 1. Department of Emergency Center, Taikang Xianlin Drum Tower Hospital, Nanjing 211800, China
    2. Department of Surgery, Nanjing Pu Kou District Hospital of Traditional Chinese Medicine General, Nanjing 211800, China
  • Received:2021-03-15 Published:2022-02-01
  • Corresponding author: Hao Ni
引用本文:

倪昊, 王发武, 李翔天, 马战胜. PSP下调miR-345-5p对雨蛙素诱导的AP腺泡细胞氧化应激和炎症因子表达[J]. 中华细胞与干细胞杂志(电子版), 2022, 12(01): 8-13.

Hao Ni, Fawu Wang, Xiangtian Li, Zhansheng Ma. Polygonatum sibiricum polysaccharide inhibits caerulein-induced oxidative stress and inflammatory factors expression of acinar cells in acute pancreatitis by down-regulating miR-345-5p[J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2022, 12(01): 8-13.

目的

探讨黄精多糖(PSP)对雨蛙素诱导的急性胰腺炎(AP)腺泡细胞氧化应激和炎症因子表达的影响及分子机制。

方法

取对数期大鼠胰腺腺泡细胞AR42J,采用100 nmol/L雨蛙素处理细胞6 h,建立AP腺泡细胞损伤模型,并采用不同浓度(1、2、4 mg/mL)PSP处理AP细胞(AP+PSP-L、AP+PSP-M、AP+PSP-H)。试剂盒检测细胞中丙二醛(MDA)含量、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GPx)活性以及培养液中白细胞介素-6 (IL-6)、肿瘤坏死因子-α (TNF-α)和IL-1β水平,实时荧光定量PCR (RT-qPCR)检测miR-345-5p表达水平。将miR-345-5p抑制物转染AR42J细胞,检测下调miR-345-5p表达对AP细胞氧化应激和炎症因子表达的影响。将miR-345-5p模拟物转染AR42J细胞,检测上调miR-345-5p表达和PSP处理对AP细胞氧化应激和炎症因子表达的影响。两组比较采用t检验,多组间比较采用方差分析,进一步两两比较采用LSD-t检验。

结果

与对照比较,AP细胞MDA含量、IL-6、TNF-α、IL-1β水平和miR-345-5p表达水平均升高,SOD和GPx活性降低(P < 0.05)。与AP细胞比较,不同浓度(1、2、4 mg/mL)PSP作用的AP细胞中MDA含量[(1.08± 0.07)比(0.88±0.06),(0.73±0.06),(0.60±0.05) nmol/mg]降低,SOD [(43.01±4.37)比(59.60±5.62),(72.37±6.32),(94.21±8.70) U/mg]和GPx活性[(29.03±2.51)比(44.11± 4.71),(58.07±4.20),(72.67± 6.56) U/mg]均升高,培养液中IL-6 [(310.72±22.27)比(257.01±20.85),(192.28±17.70),(146.93±11.90) pg/mL]、TNF-α [(223.82±21.87)比(175.57±15.85),(137.00±11.31),(89.26±7.05) pg/mL]、IL-1β表达水平[(41.66±3.85)比(33.82±3.20),(26.15±2.56),(20.14±1.71) pg/mL]和miR-345-5p表达水平(2.78±0.24比2.38±0.21,1.91±0.12,1.25±0.13)均降低,且呈浓度依赖性,差异有统计学意义(P均< 0.05)。下调miR-345-5p表达后,AP细胞中MDA含量[(1.13±0.08)比(0.72±0.06) nmol/mg]降低,SOD活性[(41.31±3.98)比(81.73±7.62) U/mg]和GPx [(28.82±2.97)比(61.41±5.81) U/mg]升高,培养液中IL-6 [(314.65±25.02)比159.76±11.93) pg/mL]、TNF-α [(235.18±23.13)比(100.41±8.09) pg/mL]和IL-1β水平[(48.67±4.50)比(27.73±2.54) pg/mL]降低(P均< 0.05)。上调miR-345-5p表达可逆转PSP对AP细胞的影响。其中MDA含量[(0.58±0.03)比(0.95±0.08) nmol/mg]升高、SOD活性[(96.52±9.54)比(54.24±4.15) U/mg]、GPx活性[(79.62±6.23)比(39.81±3.84) U/mg]降低、IL-6 [(145.38±12.49)比(275.38± 21.55) pg/mL]、TNF-α [(84.83±7.81)比(183.73±16.39) pg/mL]和IL-1β [(19.38±1.85)比(36.97±3.62) pg/mL]的表达水平升高(P均< 0.05)。

结论

PSP以剂量依赖方式减轻雨蛙素诱导AP细胞炎症反应和氧化应激损伤,其机制与下调miR-345-5p表达有关。

Objective

To study the effects of polygonatum sibiricum polysaccharide (PSP) on the oxidative stress and inflammatory factors expression in caerulein-induced acute pancreatitis (AP) acinar cells and relevant mechanisms.

Methods

Rat pancreatic acinar cells (AR42J) were treated with 100 nmol/L caerulein for 6 h to establish a model of acinar cell injury in AP. Caerulein-induced AP cells were incubated with different concentrations (1, 2, 4 mg/mL) of PSP. Malonaldehyde (MDA) content, superoxide dismutase (SOD) and glutathion peroxidase (GPx) activities in the cell, and the levels of intedeukin-6 (IL-6) , tumor necrosis factor α (TNF-α) and IL-1β in the culture medium were evaluated by commercial kits. The expression level of miR-345-5p was measured by real-time quantitative PCR (RT-qPCR) . AR42J cells were transfected with miR-345-5p inhibitors. The effect of down-regulating miR-345-5p on caerulein-induced oxidative stress and inflammatory factor expression in AP cells was examined by the above methods. The miR-345-5p mimics was transfected into AP cells, and the effects of upregulation of miR-345-5p and PSP treatment on oxidative stress and inflammatory cytokine expression was examined by the above methods. For data with homogeneity of variance, t test was used for comparison between two groups, analysis of variance for comparison between multiple groups, and LSD-t test for further pair comparison.

Results

Compared with control, MDA content, the levels of IL-6, TNF-α, IL-1β and miR-345-5p were increased in AP cells, whereas SOD and GPx activities were decreased. The MDA content [ (1.08±0.07) vs (0.88±0.06) , (0.73±0.06) , (0.60±0.05) nmol/mg] in AP cells was significantly reduced after PSP treatment, while the activities of SOD [ (43.01±4.37) vs (59.60±5.62) , (72.37±6.32) , (94.21±8.70) U/mg] and GPx [ (29.03±2.51) vs (44.11±4.71) , (58.07±4.20) , (72.67± 6.56) U/mg] were significantly increased. In addition, the expression levels of IL-6 [ (310.72±22.27) vs (257.01±20.85) , (192.28±17.70) , (146.93±11.90) pg/mL], TNF-α [ (310.72±22.27) vs (257.01±20.85) , (192.28±17.70) , (146.93±11.90) pg/mL]and IL-1β [ (41.66±3.85) vs (33.82±3.20) , (26.15±2.56) , (20.14±1.71) pg/mL] in the culture medium and the expression level of miR-345-5p (2.78±0.24 vs 2.38±0.21, 1.91±0.12, 1.25±0.13) were significantly reduced after PSP treatment in a dose-dependent manner (all P < 0.05) . After down-regulating the expression of miR-345-5p, the MDA content [ (1.13±0.08) vs (0.72±0.06) nmol/mg] in AP cells was significantly reduced, the activities of SOD [ (41.31±3.98) vs (81.73±7.62) U/mg] and GPx [ (28.82±2.97) vs (61.41±5.81) U/mg]were significantly increased, and the levels of IL-6 [ (314.65±25.02) vs (159.76±11.93) pg/mL], TNF-α [ (235.18±23.13) vs (100.41±8.09) pg/mL]and IL-1β[ (48.67±4.50) vs (27.73±2.54) pg/mL] in the culture medium were significantly reduced, with statistically significant differences (all P < 0.05) . Up-regulating miR-345-5p expression reduced the above impacts of PSP on AP cells, with an increase of expressions of MDA content [ (0.58±0.03) vs (0.95±0.08) nmol/mg], decreased SOD [ (96.52±9.54) vs (54.24±4.15) U/mg] and GPx [ (79.62±6.23) vs (39.81±3.84) U/mg] activity, as well as increased expression levels of IL-6 [ (145.38±12.49] vs (275.38±21.55) pg/mL], TNF-α [ (84.83±7.81) vs (183.73±16.39) pg/mL] and IL-1β[ (19.38±1.85) vs (36.97±3.62) pg/mL] in culture medium (P < 0.05) .

Conclusion

PSP can alleviate caerulein-induced acinar cells' inflammatory response and oxidative stress injury in acute pancreatitis in a dose-dependent manner, which is related to the down-regulation of miR-345-5p expression.

表1 引物序列信息
表2 黄精多糖对雨蛙素诱导的急性胰腺炎腺泡细胞氧化应激的影响( ± s
表3 黄精多糖对雨蛙素诱导的急性胰腺炎腺泡细胞炎症因子表达的影响( ± s
图1 黄精多糖对雨蛙素诱导急性胰腺炎腺泡细胞miR-345-5P相对表达水平的影响注:与对照比较,aP < 0.05;与AP比较,bP < 0.05;与AP+PSP-L比较,cP < 0.05;与AP+PSP-M比较,dP < 0.05
表4 抑制miR-345-5p表达对雨蛙素诱导的急性胰腺炎腺泡细胞氧化应激和炎症因子表达的影响( ± s
表5 miR-345-5p过表达逆转了黄精多糖对雨蛙素诱导的急性胰腺炎腺泡细胞氧化应激和炎症因子表达的作用( ± s
1
Cheng Z, Abrams ST, Alhamdi Y, et al. Circulating histones are major mediators of multiple organ dysfunction syndrome in acute critical illnesses[J]. Crit Care Med, 2019, 47(8):e677-e684.
2
Lankisch PG, Apte M, Banks PA. Acute pancreatitis[J]. Lancet, 2015, 386(9988):85-96.
3
贾宇涵,吴澎,郗良卿, 等. 黄精多糖提取工艺及功能作用研究进展[J]. 中国调味品, 2018, 43(11):157-161.
4
李丽,龙子江,高华武, 等. 黄精多糖对急性心肌梗死模型大鼠炎症及氧化应激反应的影响[J]. 实验动物科学, 2016, 33(5):33-38.
5
李超彦,周媛媛,王福青, 等. 黄精多糖对顺铂致肝损害大鼠肝功能的保护及抗氧化指标的影响[J]. 中国实验方剂学杂志, 2013, 19(16):229-231.
6
Cai J, Zhu Y, Zuo Y, et al. Polygonatum sibiricum polysaccharide alleviates inflammatory cytokines and promotes glucose uptake in high-glucose and high-insulin-induced 3T3-L1 adipocytes by promoting Nrf2 expression[J]. Mol Med Rep, 2019, 20(4):3951-3958.
7
刘超,徐飞,吴涛, 等. 黄精多糖对TNF-α作用的大鼠成骨细胞增殖,凋亡影响及其机制研究[J]. 中国骨质疏松杂志, 2020, 26(8):1147-1153.
8
Wan J, Yang X, Ren Y, et al. Inhibition of miR-155 reduces impaired autophagy and improves prognosis in an experimental pancreatitis mouse model[J]. Cell Death Dis, 2019, 10(4):303.
9
Kong L, Wu Q, Zhao L, et al. Effect of microRNA-27a-5p on apoptosis and inflammatory response of pancreatic acinar cells in acute pancreatitis by targeting PTEN[J]. J Cell Biochem, 2019, 120(9):15844-15850.
10
刘春雨,林宁,熊杰, 等. 高脂血症性急性胰腺炎患者血清miR-345-5p的表达及意义[J]. 解放军医学杂志, 2019, 44(4):318-321.
11
张雪良,徐子琴,熊建华. spautin-1对雨蛙素诱导的急性胰腺炎AR42J细胞凋亡及自噬的影响[J]. 中华胰腺病杂志, 2018, 18(4):247-250.
12
徐志刚,郑帅,杨泽冉, 等. NR5A2减弱雨蛙素诱导大鼠胰腺腺泡细胞的炎性反应[J]. 基础医学与临床, 2018, 38(8):1080-1087.
13
雷升萍,龙子江,施慧, 等. 黄精多糖对缺氧复氧诱导H9c2心肌细胞损伤的保护作用[J]. 中药药理与临床, 2017, 33(1):102-106.
14
Cui X, Wang S, Cao H, et al. A Review: The bioactivities and pharmacological applications of polygonatum sibiricum polysaccharides[J]. Molecules, 2018, 23(5):1170.
15
Wang Y, Lan C, Liao X, et al. Polygonatum sibiricum polysaccharide potentially attenuates diabetic retinal injury in a diabetic rat model[J]. J Diabetes Investig, 2019, 10(4):915-924.
16
Yang JX, Wu S, Huang XL, et al. Hypolipidemic activity and antiatherosclerotic effect of polysaccharide of polygonatum sibiricum in rabbit model and related cellular mechanisms[J]. Evid Based Complement Alternat Med, 2015, 2015(1):391065.
17
马怀芬,方欢乐,师西兰, 等. 黄精多糖对心脏重塑小鼠心脏组织中ICAM-1、VCAM-1蛋白表达的影响[J]. 环球中医药, 2018, 11(1):25-29.
18
Habtezion A. Inflammation in acute and chronic pancreatitis[J]. Curr Opin Gastroenterol, 2015, 31(5):395-399.
19
Yildirim AO, Ince M, Eyi YE, et al. The effects of glycyrrhizin on experimental acute pancreatitis in rats[J]. Eur Rev Med Pharmacol Sci, 2013, 17(22):2981-2987.
20
Liu TY, Zhao LL, Chen SB, et al. Polygonatum sibiricum polysaccharides prevent LPS-induced acute lung injury by inhibiting inflammation via the TLR4/Myd88/NF-κB pathway[J]. Exp Ther Med, 2020, 20(4):3733-3739.
21
Han C, Sun T, Liu Y, et al. Protective effect of polygonatum sibiricum polysaccharides on gentamicin-induced acute kidney injury in rats via inhibiting p38 MAPK/ATF2 pathway[J]. Int J Biol Macromol, 2020, 151(1):595-601.
22
Zhang Y, Yan L, Han W. Elevated level of miR-551b-5p is associated with inflammation and disease progression in patients with severe acute pancreatitis[J].Ther Apher Dial, 2018, 22(6):649-655.
23
Zhang KK, Yu SS, Li GY, et al. miR-135a deficiency inhibits the AR42J cells damage in cerulein-induced acute pancreatitis through targeting FAM129A[J]. Pflugers Arch, 2019, 471(11-12):1519-1527.
24
Ma L, Cao Y, Zhang L, et al. Celastrol mitigates high glucose-induced inflammation and apoptosis in rat H9c2 cardiomyocytes via miR-345-5p/growth arrest-specific 6[J]. J Gene Med, 2020, 22(9):e3201.
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