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北京中医药大学,北京 100029
Published:20 May 2023,
Published Online:10 February 2023,
Received:07 November 2022,
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廖文勇,李慕云,董肖等.葛花、枳椇子及其配伍对小鼠急性酒精性胃黏膜损伤的保护作用及机制[J].中国实验方剂学杂志,2023,29(10):39-47.
LIAO Wenyong,LI Muyun,DONG Xiao,et al.Protective Effect of Flos Puerariae, Hoveniae Semen, and Their Compatibility on Acute Alcoholic Gastric Mucosal Injury in Mice and Mechanism[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(10):39-47.
廖文勇,李慕云,董肖等.葛花、枳椇子及其配伍对小鼠急性酒精性胃黏膜损伤的保护作用及机制[J].中国实验方剂学杂志,2023,29(10):39-47. DOI: 10.13422/j.cnki.syfjx.20230243.
LIAO Wenyong,LI Muyun,DONG Xiao,et al.Protective Effect of Flos Puerariae, Hoveniae Semen, and Their Compatibility on Acute Alcoholic Gastric Mucosal Injury in Mice and Mechanism[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(10):39-47. DOI: 10.13422/j.cnki.syfjx.20230243.
目的
2
探讨葛花、枳椇子及其配伍对急性酒精性胃黏膜损伤的改善作用,为进一步开展葛花、枳椇子及其配伍防治酒精致多脏器损伤奠定基础。
方法
2
采用多次灌胃给予56%红星二锅头白酒(15 mL·kg
-1
)建立小鼠急性酒精性胃黏膜损伤模型,将120只ICR雄性小鼠随机分为空白组、模型组、奥美拉唑组(0.026 g·kg
-1
)、葛花-枳椇子(配伍)高、中、低剂量组(29.2、14.6、7.3 g·kg
-1
)、葛花组(19.5 g·kg
-1
)、枳椇子组(19.5 g·kg
-1
)共8个组,每组15只,动物适应性喂养1周后,按10 mL·kg
-1
预给相应药物3 d,从第4天开始,给药1 h后按15 mL·kg
-1
灌胃二锅头白酒,空白组给予相同体积去离子水,记录小鼠醉酒和醒酒时间,连续给药给酒3 d,末次给药1 h后摘眼球处死;气相色谱仪测定各组小鼠血清中乙醇体积分数,紫外-可见分光光度计检测各组小鼠胃黏膜中乙醇脱氢酶(ADH)活性;苏木素-伊红(HE)染色观察胃黏膜病理变化;酶联免疫吸附测定法(ELISA)检测各组小鼠血清中炎症因子含量;实时荧光定量聚合酶链式反应(Real-time PCR)检测核转录因子-
κ
B(NF-
κ
B)p65和NF-
κ
B抑制蛋白
α
(I
κ
B
α
)mRNA表达。
结果
2
与正常组比较,模型组血清中白细胞介素-6(IL-6)、白细胞介素-1
β
(IL-1
β)
和肿瘤坏死因子-
α
(TNF-
α)
含量升高(
P
<
0.05),胃黏膜组织NF-
κ
B p65 mRNA表达升高(
P
<
0.01),I
κ
B
α
mRNA表达降低(
P
<
0.01);与模型组比较,奥美拉唑组、配伍高、中剂量组、葛花组醉酒时间延长(
P
<
0.05),配伍高、中剂量组醒酒时间缩短(
P
<
0.05),配伍高剂量组血清中乙醇体积分数降低(
P
<
0.05),奥美拉唑组、配伍高、中剂量组胃黏膜中ADH活性升高(
P
<
0.05),配伍各剂量组、葛花组肉眼损伤评分降低(
P
<
0.05),奥美拉唑组、配伍各剂量组、葛花组病理损伤评分降低(
P
<
0.01),各给药组血清中IL-6表达降低(
P
<
0.05),奥美拉唑组、配伍各剂量组、枳椇子组血清中IL-1
β
表达降低(
P
<
0.05),配伍高、中剂量组血清中TNF-
α
表达降低(
P
<
0.05),各给药组胃黏膜组织NF-
κ
B p65 mRNA表达降低(
P
<
0.05),奥美拉唑组、配伍各剂量组胃黏膜组织I
κ
B
α
mRNA表达增加(
P
<
0.05);与高剂量组比较,配伍低剂量组与枳椇子组醉酒时间缩短(
P
<
0.01),葛花、枳椇子组醒酒时间延长(
P
<
0.01),配伍中、低剂量组、葛花组、枳椇子组血清中乙醇体积分数升高(
P
<
0.05),配伍中、低剂量组、枳椇子组肉眼损伤积分增加(
P
<
0.05),配伍中、低剂量组、葛花组、枳椇子组病理损伤积分增加(
P
<
0.01),配伍低剂量组、葛花组、枳椇子组血清中IL-1
β
含量升高(
P
<
0.01),葛花组与枳椇子组胃黏膜组织I
κ
B
α
mRNA表达量降低(
P
<
0.05);与配伍中剂量组比较,枳椇子组醉酒时间缩短(
P
<
0.05),葛花组醒酒时间延长(
P
<
0.05),葛花组、枳椇子组病理损伤积分增加(
P
<
0.01),配伍低剂量组、葛花组、枳椇子组血清中IL-1
β
含量升高(
P
<
0.05);与配伍低剂量组比较,枳椇子组病理损伤积分增加(
P
<
0.05)。
结论
2
葛花、枳椇子及其配伍能起到对小鼠急性酒精性胃黏膜损伤的防治作用,可能与抑制胃黏膜NF-
κ
B信号通路的表达有关,且配伍高剂量组药效最佳。
Objective
2
To explore the improvement effect of Flos Puerariae, Hoveniae Semen, and their compatibility on acute alcoholic gastric mucosal injury, and lay a foundation for further development of Flos Puerariae, Hoveniae Semen, and their compatibility in the prevention and treatment of alcohol-induced multiple organ injury.
Method
2
The acute alcohol-induced gastric mucosal injury model of mice was established by multiple intragastric administration of 56% Hongxing Erguotou liquor (15 mL·kg
-1
). A total of 120 male ICR mice were randomly divided into 8 groups, namely, the blank group, model group, omeprazole group (0.026 g·kg
-1
), Flos Puerariae-Hoveniae Semen (compatibility) high, medium, and low-dose groups (29.2,14.6, 7.3 g·kg
-1
), Flos Puerariae group (19.5 g·kg
-1
), and Hoveniae Semen group (19.5 g·kg
-1
), with 15 mice in each group. After one week of adaptive feeding, the animals were pre-administrated with the corresponding drug at the rate of 10 mL·kg
-1
for 3 d. From the 4
th
day, after 1 h of administration, Erguotou liquid was administrated at the rate of 15 mL·kg
-1
and the blank group was administrated with the same volume of deionized water to record the drunkenness and sober up time. The administration was lasted for 3 d. One hour after the last administration, the eyeballs were removed and the mice were sacrificed. The concentration of ethanol in serum was determined by gas chromatograph, and the activity of ethanol dehydrogenase (ADH) in gastric mucosa was determined by ultraviolet-vis spectrophotometer. Hematoxylin-eosin (HE) staining was used to observe the pathological changes in gastric mucosa. Serum inflammatory factors were determined by enzyme-linked immunosorbent assay (ELISA). The mRNA expression of nuclear transcription factor-
κ
B (NF-
κ
B) p65 and NF-
κ
B inhibitory protein
α
(I
κ
B
α
) were detected by real-time polymerase chain reaction (Real-time PCR).
Result
2
As compared with the normal group, the content of interleukin-6 (IL-6), interleukin-1
β
(IL-1
β
), and tumor necrosis factor-
α
(TNF-
α
) in serum of mice in the model group was increased (
P
<
0.05), the mRNA expression of NF-
κ
B p65 in gastric mucosa tissues was increased (
P
<
0.01), and the mRNA expression of I
κ
B
α
was decreased (
P
<
0.01). As compared with the model group, the drunkenness time of the omeprazole group, high and medium-dose compatibility groups, and Flos Puerariae group was prolonged (
P
<
0.05), the sober up time of the high and medium-dose compatibility groups was shortened (
P
<
0.05), the ethanol concentration in the serum of the high-dose compatibility group was decreased (
P
<
0.05), the ADH activity in the gastric mucosa of the omeprazole group and high and medium-dose compatibility groups was increased (
P
<
0.05), the macroscopic injury score of the high, medium, and low-dose compatibility groups and Flos Puerariae group was decreased (
P
<
0.05), the score of pathological injury in the omeprazole group, high, medium, and low-dose compatibility groups, and Flos Puerariae group was decreased (
P
<
0.01), the expression of IL-6 in serum of all drug groups was decreased (
P
<
0.05), the expression of IL-1
β
in serum of the omeprazole group, high, medium, and low-dose Flos Puerariae groups, and Hoveniae Semen group was decreased (
P
<
0.05), the expression of TNF-
α
in serum of high and medium-dose groups was decreased (
P
<
0.05), the mRNA expression of NF-
κ
B p65 in gastric mucosa tissues of all drug groups was decreased (
P
<
0.05), and the mRNA expression of I
κ
B
α
in gastric mucosa tissues of the omeprazole group and high, medium, and low-dose compatibility groups was increased (
P
<
0.05). As compared with the high-dose compatibility group, the drunkenness time in the low-dose compatibility group and Hoveniae Semen group was shortened (
P
<
0.01), the sober up time in the Flos Puerariae and Hoveniae Semen groups was prolonged (
P
<
0.01), the concentration of ethanol in the serum of the medium and low-dose compatibility groups, Flos Puerariae group, and Hoveniae Semen group increased (
P
<
0.05), the macroscopic injury score of the medium and low-dose compatibility groups and Hoveniae Semen group was increased (
P
<
0.05), the pathological injury score of the medium and low-dose compatibility groups, Flos Puerariae group, and Hoveniae Semen group was increased (
P
<
0.01), the content of IL-1
β
in serum of low-dose compatibility group, Flos Puerariae group, and Hoveniae Semen group was increased (
P
<
0.01), and the mRNA expression of I
κ
B
α
in gastric mucosa of the Flos Puerariae group and Hoveniae Semen group was decreased (
P
<
0.05). As compared with the medium-dose compatibility group, the drunkenness time in the Hoveniae Semen group was shortened (
P
<
0.05), the sober up time in the Flos Puerariae group was prolonged (
P
<
0.05), the pathological injury score in the Flos Puerariae group and Hoveniae Semen group was increased (
P
<
0.01), and the content of IL-1
β
in serum of the low-dose compatibility group, the Flos Puerariae group, and Hoveniae Semen group was increased (
P
<
0.05). As compared with the low-dose compatibility group, the pathological injury score of the Hoveniae Semen group was increased (
P
<
0.05).
Conclusion
2
Flos Puerariae, Hoveniae Semen, and their compatibility play a role in preventing and treating acute alcoholic gastric mucosal injury in mice, which may be related to the inhibition of the expression of NF-
κ
B signal pathway in gastric mucosa, and the high-dose compatibility group has the optimal effect.
葛花枳椇子急性胃黏膜损伤乙醇代谢炎症反应
Flos PuerariaeHoveniae Semenacute gastric mucosal injuryethanol metabolisminflammatory reaction
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