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加味膈下逐瘀汤通过 p38 信号通路缓解慢性输卵管炎。

Modified Gexiazhuyu decoction alleviates chronic salpingitis p38 signaling pathway.

机构信息

Second Affiliated Hospital of Heilongjiang University of Chinese Medicine, Heilongjiang 150001, China.

Heilongjiang University of Traditional Chinese Medicine, Heilongjiang 150040, China.

出版信息

J Tradit Chin Med. 2022 Apr;42(2):213-220. doi: 10.19852/j.cnki.jtcm.2022.02.003.

Abstract

OBJECTIVE

To investigate pharmacodynamic effects of modified Gexiazhuyu decoction (MGXZYD) and explore the underlying mechanism in the treatment of chronic salpingitis METHODS: Chronic salpingitis model rats were firstly constructed and the blood was collected to detect the whole blood viscosity and plasma viscosity. Rat oviduct were collected to evaluate the macroscopic damage and the pathological injury and fibrosis of oviduct by hematoxylin-eosin (HE) and Masson staining. Elisa assay was to detect the production interleukin-1 β (IL-1β) in serum and collagen I (COL-1), matrix metalloprotein 9 (MMP-9), tissue inhibitor of metalloproteinases 1 (TIMP-1) in oviduct tissue. And immunohistochemical staining with MMP-9 and TIMP-1 in oviduct tissue were examined. Western blot was used to detect the expressions of p38 mitogen-activated protein kinases (p38MAPK), phospho-p38MPAK (p-p38MPAK), transforming growth factor-β1 (TGF-β1) in oviduct. The expression of α-smooth muscle actin (α-SMA), p-p38MPAK, in oviduct tissue were detected by immunofluorescence method. The mRNA of p-p38MAPK, α -SMA, COL-1, MMP-9, TIMP-1 was measured by reverse transcription-polymerase chain reaction.

RESULTS

Rats administrated with MGXZYD demonstrated decreased the whole blood viscosity and plasma viscosity. MGXZYD obviously improved the tubal wall thickening, swelling and pelvic adhesion. And HE and Masson staining showed MGXZYD improved the pathological injury and fibrosis of oviduct. The results of MTT assay and flow cytometry indicated that MGXZYD could decreased the NIN-3T3 cells viability and improved the apoptosis. Besides, MGXZYD inhibited the protein and / or mRNA of TGF-β1, IL-1β, COL-1, α-SMA, p-p38MAPK expressions and increased the production of MMP-9/TIMP-1.

CONCLUSION

MGXZYD could prevent the progression of chronic salpingitis by inhibited the fibrocyte and inflammation which inhibited the p38 MAPK signaling pathway.

摘要

目的

研究加味膈下逐瘀汤(MGXZYD)的药效学作用,并探讨其治疗慢性输卵管炎的作用机制。

方法

首先构建慢性输卵管炎模型大鼠,采集全血检测全血黏度和血浆黏度。采集大鼠输卵管,通过苏木精-伊红(HE)和 Masson 染色评估输卵管的大体损伤和组织病理学损伤及纤维化。酶联免疫吸附试验(ELISA)检测血清中白细胞介素-1β(IL-1β)、输卵管组织中胶原 I(COL-1)、基质金属蛋白酶 9(MMP-9)、基质金属蛋白酶组织抑制剂 1(TIMP-1)的产生。检测输卵管组织中 MMP-9 和 TIMP-1 的免疫组织化学染色。Western blot 检测输卵管组织中 p38 丝裂原活化蛋白激酶(p38MAPK)、磷酸化 p38MAPK(p-p38MAPK)、转化生长因子-β1(TGF-β1)的表达。免疫荧光法检测输卵管组织中α-平滑肌肌动蛋白(α-SMA)、p-p38MAPK 的表达。采用逆转录-聚合酶链反应(RT-PCR)法检测 p-p38MAPK、α-SMA、COL-1、MMP-9、TIMP-1 的 mRNA 表达。

结果

给予 MGXZYD 的大鼠全血黏度和血浆黏度降低。MGXZYD 明显改善了输卵管壁增厚、肿胀和盆腔粘连。HE 和 Masson 染色显示,MGXZYD 改善了输卵管的组织病理学损伤和纤维化。MTT 检测和流式细胞术结果表明,MGXZYD 可降低 NIN-3T3 细胞活力,促进细胞凋亡。此外,MGXZYD 抑制 TGF-β1、IL-1β、COL-1、α-SMA、p-p38MAPK 的蛋白和/或 mRNA 表达,并增加 MMP-9/TIMP-1 的产生。

结论

MGXZYD 通过抑制成纤维细胞和炎症反应,抑制 p38MAPK 信号通路,可防止慢性输卵管炎的进展。

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