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1.江西中医药大学 中药固体制剂制造技术国家工程研究中心,南昌 330006
2.江西中医药大学 药学院,南昌 330004
王玲玲,硕士,从事中药药效评价与新药研究,E-mail:2584032268@qq.com
陈兰英,博士,教授,从事中药药效评价与新药研究,E-mail:clyxy2513@163.com
修回日期:2019-12-19,
网络出版日期:2020-05-27,
纸质出版日期:2020-08-05
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王玲玲,崔亚茹,陈兰英等.白头翁皂苷B4对慢性子宫内膜炎大鼠的治疗作用及其相关因子的影响[J].中国实验方剂学杂志,2020,26(15):54-60.
WANG Ling-ling,CUI Ya-ru,CHEN Lan-ying,et al.Therapeutic Effect of Anemoside B4 on Chronic Endometritis in Rats and Its Related Factors[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(15):54-60.
王玲玲,崔亚茹,陈兰英等.白头翁皂苷B4对慢性子宫内膜炎大鼠的治疗作用及其相关因子的影响[J].中国实验方剂学杂志,2020,26(15):54-60. DOI: 10.13422/j.cnki.syfjx.20201537.
WANG Ling-ling,CUI Ya-ru,CHEN Lan-ying,et al.Therapeutic Effect of Anemoside B4 on Chronic Endometritis in Rats and Its Related Factors[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(15):54-60. DOI: 10.13422/j.cnki.syfjx.20201537.
目的
2
通过体内外实验研究白头翁皂苷B
4
对子宫内膜炎大鼠的影响。
方法
2
动物实验采用25%的苯酚胶浆制备子宫内膜炎模型,60只SD雌性大鼠随机分为空白组、模型组、苦参凝胶组(0.005 g·kg
-1
),白头翁皂苷B
4
凝胶低、中、高剂量组(0.005,0.01,0.02 g·kg
-1
),每组10只,除空白组外,各组大鼠每2 d向其阴道内注入25%苯酚胶浆,造模持续30 d。造模后当天开始给药,白头翁皂苷B
4
凝胶低、中、高剂量组每日直肠给药,苦参凝胶组每日阴道给药,空白组和模型组以同样的方法给予等量的生理盐水,给药连续30 d。末次给药后,取各组大鼠子宫及其附件,进行大鼠子宫形态、子宫指数分析;苏木素-伊红(HE)染色观察大鼠子宫组织病理变化;实时荧光定量聚合酶链式反应(Real-time PCR)检测各组大鼠子宫组织中肿瘤坏死因子-
α
(TNF-
α
),白细胞介素-1
β
(IL-1
β
),白细胞介素-6(IL-6),信号转导蛋白130(gp130),信号传导及转录激活因子3(STAT3)mRNA的表达;蛋白免疫印迹法(Western blot)检测大鼠子宫组织中IL-6和STAT3蛋白的表达;细胞实验,以脂多糖(LPS)诱导大鼠子宫内膜上皮细胞制备体外炎症模型,Real-time PCR检测各组大鼠子宫内膜上皮细胞中IL-6,gp130和STAT3 mRNA的表达。
结果
2
动物实验结果显示,与空白组比较,模型组大鼠子宫宫腔黏连不全,子宫内膜出现水肿及充血病变;与模型组比较,白头翁皂苷B
4
凝胶高剂量组和苦参凝胶组大鼠子宫宫腔无黏连现象,子宫组织较为完整,子宫病理结构得到明显的改善。与空白组比较,模型组大鼠子宫指数明显升高(
P
<
0.05),子宫组织中IL-1
β
mRNA表达明显升高(
P
<
0.05)
,
子宫组织中IL-6,STAT3的mRNA和蛋白表达明显升高(
P
<
0.05);与模型组比较,白头翁皂苷B
4
凝胶高剂量组中子宫指数明显降低(
P
<
0.05),子宫组织中IL-6,STAT3的mRNA和蛋白表达水平明显降低(
P
<
0.05),对TNF-
α
和IL-1
β
的mRNA表达与模型组比较差异无统计学意义。细胞实验结果显示,与空白组比较,模型组大鼠子宫内膜上皮细胞中IL-6和gp130的mRNA表达显著升高(
P
<
0.01),STAT3的mRNA表达明显升高(
P
<
0.05);与模型组比较,白头翁皂苷B
4
高剂量组中IL-6,gp130和STAT3的mRNA表达水平明显下降(
P
<
0.05)。
结论
2
白头翁皂苷B
4
可改善大鼠慢性子宫内膜炎的炎症反应,降低炎症因子IL-6的释放,其机制可能与IL-6/STAT3通路的下调有关。
Objective
2
The effects of anemoside B
4
on endometritis rats were studied through
in vivo
and
in vitro
experiments.
Method
2
Animal experiments used 25% phenol glue to prepare endometritis models. 60 female SD rats were randomly divided into blank group, model group, Kushen gel group(0.005 g·kg
-1
),anemoside B
4
gel low,medium and high dose groups(0.005,0.01,0.02 g·kg
-1
),10 rats in each group,except for the blank group,rats in each group were injected with 25% phenol glue into their vagina every 2 days,and the modeling continued for 30 days. Administration started on the day after modeling. Anemoside B
4
gel low, medium and high dose groups were administered rectal daily,Kushen gel group was given daily vaginal administration. The blank group and model group were given the same amount of normal saline in the same way for 30 consecutive days. After the last administration,the uterus and its attachments of each group of rats were taken to analyze the uterine morphology and index. Hematoxylin-eosin (HE) staining was used to observe the pathological changes of rat uterus. Real-time PCR was used to detect tumor necrosis factor-
α
(TNF-
α
),interleukin-1
β
(IL-1
β
),and interleukin-6 (IL-6),signal transduction protein 130 (gp130),signal transduction and transcription activator 3 (STAT3)mRNA expression. Detection of IL-6 and STAT3 protein expression in rat uterus by Western blot. In cell experiments,lipopolysaccharide (LPS)was used to induce rat endometrial epithelial cells to prepare an in vitro inflammation model, and Real-time PCR was used to detect the expression of IL-6,gp130 and STAT3 mRNA in each group of rat endometrial epithelial cells.
Result
2
The results of animal experiments showed that compared with the blank group, the model group had inadequate uterine cavity adhesions, endometrial edema and hyperemia. Compared with model group, there was no adhesion in the uterine cavity of the rats in the high dose anemoside B
4
gel group and the Kushen gel group. The uterine tissue was relatively complete, and the uterine pathological structure was significantly improved. Compared with the blank group, the uterine index of the model group was significantly increased(
P
<
0.05), the expression of IL-1
β
mRNA in the uterine tissue was significantly increased (
P
<
0.05), the expression of mRNA and protein of IL-6 and STAT3 in the uterine tissue significantly increased (
P
<
0.05). Compared with model group, the uterine index in anemoside B
4
gel high dose group was significantly reduced (
P
<
0.05), and the mRNA and protein expression levels of IL-6 and STAT3 in the uterine tissue were significantly reduced (
P
<
0.05). There was no statistically significant difference between the TNF-
α
and IL-1
β
mRNA expression compared with the model group. Cell experiment results showed that compared with the blank group, the mRNA expression of IL-6 and gp130 in model group endometrial epithelial cells was significantly increased (
P
<
0.01), STAT3 mRNA expression was significantly increased (
P
<
0.05). Compared with model group, the mRNA expression levels of IL-6, gp130 and STAT3 in anemoside B
4
high dose group decreased significantly (
P
<
0.05).
Conclusion
2
Anemoside B
4
can improve the inflammatory response of chronic endometritis in rats and reduce the release of inflammatory factor IL-6. The mechanism may be related to the down-regulation of IL-6/STAT3 pathway.
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