益母草总生物碱对药物流产大鼠蜕膜绒毛组织排出的促进作用及其机制

doi:10.13481/j.1671-587X.20210618

摘要/Abstract

摘要： 目的

探讨益母草总生物碱对药物流产大鼠蜕膜绒毛组织排出的促进作用，并阐述其可能的作用机制。

方法

选取动情期健康雌性SD大鼠100只和雄性SD大鼠50只合笼，在受孕雌鼠中选取60只。随机选取9只作为正常组，其余51只大鼠建立药物流产后子宫复旧不全病理模型，成功建模的40只大鼠随机分为模型组、激活剂组、激活剂+益母草总生物碱组和益母草总生物碱组（n=10）分别给予生理盐水、磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（AKT）信号通路激活剂胰岛素样生长因子1（IGF-1）、IGF-1+益母草总生物碱和益母草总生物碱灌胃。正常组大鼠灌胃等体积生理盐水。检测各组大鼠子宫出血量、凝血时间、子宫质量、子宫系数、子宫平滑肌张力和收缩力，HE染色观察大鼠子宫组织病理形态表现，免疫组织化学法检测各组大鼠子宫内膜组织中雌激素受体（ER）蛋白表达水平，Western bloting法检测各组大鼠蜕膜组织中p38丝裂原活化蛋白激酶（p38MAPK）、PI3K、磷酸化PI3K（p-PI3K）、AKT和磷酸化AKT（p-AKT）蛋白表达水平。

结果

与正常组比较，激活剂组、模型组、激活剂+益母草碱组和益母草碱组大鼠子宫出血量增加（P<0.05），凝血时间缩短（P<0.05），子宫质量和子宫系数降低（P<0.05），子宫平滑肌张力和收缩力增大（P<0.05），大鼠子宫蜕膜组织中ER表达水平升高（P<0.05），子宫蜕膜组织中p-p38MAPK/p38MAPK、p-PI3K/PI3K和p-AKT/AKT比值降低（P<0.05）；与激活剂组比较，模型组、激活剂+益母草总生物碱组和益母草总生物碱组大鼠子宫出血量增加（P<0.05），凝血时间延长（P<0.05），大鼠子宫质量和子宫系数降低（P<0.05），子宫平滑肌张力与收缩力增大（P<0.05），大鼠子宫蜕膜组织中ER表达水平升高（P<0.05），子宫蜕膜组织中p-p38MAPK/p38MAPK、p-PI3K/PI3K和p-AKT/AKT比值降低（P<0.05）；与模型组比较，激活剂+益母草总生物碱组和益母草总生物碱组大鼠子宫出血量增加（P<0.05），凝血时间延长（P<0.05），大鼠子宫质量和子宫系数降低（P<0.05），子宫平滑肌张力与收缩力增大（P<0.05），大鼠子宫蜕膜组织中ER表达水平升高（P<0.05），子宫蜕膜组织中p-p38MAPK/p38MAPK、p-PI3K/PI3K和p-AKT/AKT比值降低（P<0.05）；与激活剂+益母草总生物碱组比较，益母草总生物碱组大鼠子宫出血量降低（P<0.05），凝血时间延长（P<0.05），大鼠子宫质量和子宫系数降低（P<0.05），子宫平滑肌张力与收缩力增大（P<0.05），大鼠子宫蜕膜组织中ER表达水平升高（P<0.05），子宫蜕膜组织中p-p38MAPK/p38MAPK、p-PI3K/PI3K和p-AKT/AKT比值降低（P<0.05）。激活剂组和模型组大鼠子宫内有大量淤血和蜕膜细胞，激活剂+益母草总生物碱组和益母草总生物碱组大鼠子宫无淤血，蜕膜细胞明显减少。

结论

益母草总生物碱可促进药物流产大鼠蜕膜绒毛组织排出，其作用机制可能与抑制MAPK/PI3K/AKT信号通路有关。

关键词: 益母草总生物碱, 丝裂原活化蛋白激酶/磷脂酰肌醇3-激酶/蛋白激酶B信号通路, 药物流产, 蜕膜组织

Abstract: Objective

To investigate the promotion effect of total alkaloids of leonurus on decidual villus excretion in the drug abortion rats， and to explore its possible mechanism.

Methods

A total of 100 healthy female SD rats during estrus and 50 male SD rats were caged， and 60 pregnant female rats were selected. Nine rats were randomly selected as normal group， and the other 51 rats were used to establish the pathological models of incomplete uterine involution after drug abortion. A total of 40 successfully modeled rats were randomly divided into model group， activator group， activator +leonurus alkaloid group and leonurus alkaloid group（n=10），and the rats were given phosphatidylinositol 3-kinase （PI3K）/protein kinase B （AKT） signaling pathway activator insulin-like growth factor-1 （IGF-1）， PI3K/AKT activator IGF-1+total alkaloids of leonurus and total alkaloids of leonurus by gavage， and the rats in normal group were given equal volume of normal saline by gavage. The bleeding volume， coagulation time， wet weight，and coefficient of the urerus， tension and contractility of uterus smooth muscle of the rats in various groups were detected. The morphology of uterine tissue of the rats in various groups was observed by HE staining， the expression levels of estrogen receptor （ER） in decidual tissue of the rats in various groups were detected by immunohistochemistry， and Western blotting method was used to detect the expression levels of p38 mitogen activated protein kinase （p38MAPK），PI3K，phosphorylated PI3K（p-PI3K），AKT and phosphorylated AKT （p-AKT） proteins in decidual tissue of uterus of the rats in various groups.

Results

Compared with normal group， the bleeding volume， coagulation time， weight and coefficient of uterus of the rats in activator group， model group， activator+leonurus alkaloid group and leonurus alkaloid group were decreased （P<0.05）， the tension and contractility of uterine smooth muscle were increased （P<0.05）， the expression levels of ER in decidual tissue of uterus were increased （P<0.05）， and the ratios of P-P38MAPK / p38MAPK， p-PI3K/PI3K and p-AKT/AKT in decidual tissue of uterus were decreased （P<0.05）； compared with activator group， the bleeding volume， coagulation time， uterus weights and uterus coefficients of the rats in model group， activator+leonurus alkaloid group and leonurus alkaloid group were increased （P<0.05）， the tensions and contractilities of uterine smooth muscle were increased （P<0.05）， and the expression levels of ER in decidual tissue of uterus were increased （P<0.05） and the ratios of p-P38MAPK / p38MAPK， p-PI3K / PI3K and p-AKT/AKT in decidual tissue of uterus were decreased （P<0.05）；compared with model group， the bleeding volume of uterus of the rats in activator+leonurus alkaloid group and leonurus alkaloid group were decreased （P<0.05）， the coagulation time was prolonged， the tensions and contractility of the uterine smooth muscle were increased （P<0.05），the expression levels of ER in decidual tissue of the uterus were increased （P<0.05）， and the ratios of p-P38MAPK/p38MAPK， p-PI3K/PI3K and p-AKT/AKT in decidual tissue of uterus were decreased （P<0.05）； compared with activator+leonurus alkaloid group， the bleeding volume of uterus of the rats in leonurus alkaloid group was decreased （P<0.05）， the coagulation time was prolonged（P<0.05），the tension and contractility of uterine smooth muscle were increased （P<0.05）， and the expression level of ER in decidual tissue of uterus was increased（P<0.05），and the ratios of p-p38MAPK/p38MAPK，p-PI3K/PI3K and p-AKT/AKT in decidual tissue of uterus were decreased （P<0.05）.There were a large amount of congestion and decidual cells in the rats in activator group and model group. There was no congestion and decidual cells decreased significantly in the activator+leonurus alkaloid group and leonurus alkaloid group.

Conclusion

The total alkaloids of leonurus can promote the excretion of decidual villi tissue of the drug abortion rats， which may be achieved by inhibiting the MAPK/PI3-K/AKT signaling pathway.

Key words: total alkaloids of leonurus, mitogen activated protein kinase/phosphatidylinositol 3-kinase/protein kinase B signaling pathway, drug abortion, decidual tissue

中图分类号:

R285.5

马静静,刘宇,梁艳,李颖. 益母草总生物碱对药物流产大鼠蜕膜绒毛组织排出的促进作用及其机制[J]. 吉林大学学报(医学版), 2021, 47(6): 1476-1484.

Jingjing MA,Yu LIU,Yan LIANG,Ying LI. Promotion effect of total alkaloids of leonurus on decidual villus excretion in drug abortion rats and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2021, 47(6): 1476-1484.

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参考文献 20

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Group	n	bleeding volume(V/mL)	Coagulation time(t/s)
Normal	9	0.03±0.01	82.24±11.78
Activator	10	0.46±0.07^*	45.64±4.31^*
Model	10	0.40±0.04^*△	51.58±3.97^*△
Activator+leonurus alkaloid	10	0.29±0.03^*△#	62.34±3.28^*△#
Leonurus alkaloid	10	0.17±0.01^*△○	71.56±3.05^*△○
F		185.410	57.489
P		<0.001	<0.001

Group	n	Uterus weight (m/g)	Uterus coefficient
Normal	9	6.32±1.03	0.022 5±0.003 2
Activator	10	0.88±0.09^*	0.003 5±0.000 9^*
Model	10	0.56±0.06^*△	0.002 4±0.000 5^*△
Activator+leonurus alkaloid	10	0.43±0.04^*△#	0.001 6±0.000 2^*△#
Leonurus alkaloid	10	0.32±0.03^*△#○	0.000 9±0.000 1^*△#○
F		312.780	370.405
P		<0.001	<0.001

Group	n	Tension	Contractility
Normal	9	1.49±0.31	5.12±1.10
Activator	10	4.68±0.14^*	29.64±4.66^*
Model	10	5.81±0.24^*△	48.55±5.31^*△
Activator+leonurus alkaloid	10	6.33±0.25^*△#	56.47±4.95^*△#
Leonurus alkaloid	10	7.22±0.19^*△#○	65.48±3.91^*△#○
F		860.231	293.857
P		<0.001	<0.001

Group		ER
Normal		29.43±2.11
Activator		17.61±1.13^*
Model		20.34±0.97^*△
Activator+leonurus alkaloid		23.48±1.31^*△#
Leonurus alkaloid		27.06±1.23^*△#○
F		35.816
P		<0.001

Group	p-p38MAPK/p38MAPK	p-PI3-K/PI3-K	p-AKT/AKT
Normal	87.42±4.64	79.94±3.28	95.31±5.31
Activator	37.48±1.71^*	35.15±1.23^*	36.34±1.45^*
Model	25.15±1.32^*△	28.75±1.25^*△	29.64±1.72^*△
Activator+leonurus alkaloid	19.67±1.52^*△#	14.39±0.76^*△#	16.39±1.25^*△#
Leonurus alkaloid	11.36±0.58^*△#○	9.64±0.12^*△#○	10.36±1.36^*△#○
F	472.928	810.117	469.122
P	<0.001	<0.001	<0.001