Mechanism study of Qingchang mixture in the treatment of postoperative abdominal adhesions by regulating the expression of Th1 and Th2 cytokines

doi:10.11958/20241146

Abstract

Abstract:

Objective To elucidate the efficacy and possible mechanism of Qingchang mixture in ameliorating postoperative abdominal adhesions in rats. Methods Seventy-five Sprague-Dawley rats were randomly allocated into five experimental groups: the control group, the sham-operated group, the model group, the Qingchang mixture treatment group and the sodium hyaluronate treatment group. Except the control group and the sham-operated group, the other three groups were treated with cecal abrasion method to establish the rat model of abdominal adhesion. In the sodium hyaluronate group, 2 mL sodium hyaluronate gel was meticulously applied to the injured abdominal wall and cecum prior to abdominal closure. Following successful establishment of adhesion model, the Qingchang mixture group received a daily oral gavage of 2 mL Qingchang mixture (14.58 g/kg), while the other four groups were given equal volume normal saline administration. In each group, five rats were euthanized on postoperative days 3, 7 and 14 to assess abdominal adhesions using Nair’s scoring system. Adhesive tissue or normal peritoneal tissue were harvested on postoperative day 7, and mRNA expression levels of interleukin-4 (IL-4), signal transducer and activator of transcription 6 (STAT-6) and interferon-γ (IFN-γ) were quantified via fluorescence-based real-time PCR. Concurrently, Western blot analysis was employed to evaluate protein expression levels of IL-4, interleukin-10 (IL-10), STAT-6 and IFN-γ. Pathological alterations in adhesive tissue were visualized using hematoxylin-eosin (HE) staining under light microscopy, and inflammation and fibrotic changes were assessed accordingly. Results Compared with the blank and sham-operated groups, mRNA and protein expression levels of IL-4 and STAT-6 were significantly downregulated in the model group, protein expression level of IL-10 was also reduced. Conversely, the mRNA and protein expression levels of IFN-γ, as well as the inflammation and fibrosis scores were significantly elevated (P<0.05). In comparison to the model group, IL-4 and STAT-6 mRNA and protein expression levels were increased in the Qingchang mixture group and the hyaluronic acid group, along with an increase in IL-10 protein expression. Conversely, these groups exhibited a significant reduction in Nair’s scores, inflammation scores, fibrosis scores, and IFN-γ mRNA and protein expression levels were decreased (P<0.05). Conclusion Qingchang mixture appears to suppress the development of postoperative peritoneal adhesions, likely through mechanism that involves modulating the expression of T-helper 1 and T-helper 2 cytokines, thereby attenuating inflammatory response.

Key words: Th1 cells, Th2 cells, postoperative intra-abdominal adhesions, Qingchang mixture, inflammatory response

CLC Number:

R285.5，R656

Figures/Tables 8

References 24

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基因名称	引物序列（5′→3′）	产物大小/bp
IFN-γ	上游：GAGGAACTGGCAAAAGGACG	132
IFN-γ	下游：AGGTGCGATTCGATGACACT	132
IL-4	上游：TGTAGAGGTGTCAGCGGTCT	70
IL-4	下游：TCAGTGTTGTGAGCGTGGAC	70
STAT-6	上游：TCAGCACCGAGTAAGGGAGA	130
STAT-6	下游：GGCCAGATCCTCACTTGGAC	130
β-actin	上游：CTCTGIGTGGATTGGTGGCT	139
β-actin	下游：CGCAGCTCAGTAACAGTCCG	139

基因名称	引物序列（5′→3′）	产物大小/bp
IFN-γ	上游：GAGGAACTGGCAAAAGGACG	132
IFN-γ	下游：AGGTGCGATTCGATGACACT	132
IL-4	上游：TGTAGAGGTGTCAGCGGTCT	70
IL-4	下游：TCAGTGTTGTGAGCGTGGAC	70
STAT-6	上游：TCAGCACCGAGTAAGGGAGA	130
STAT-6	下游：GGCCAGATCCTCACTTGGAC	130
β-actin	上游：CTCTGIGTGGATTGGTGGCT	139
β-actin	下游：CGCAGCTCAGTAACAGTCCG	139

组别	0	Ⅰ	Ⅱ	Ⅲ	Ⅳ	Nair’s评分/分
假手术组	9	4	2	0	0	1.28±0.25
模型组	0	3	6	5	1	2.27±0.42^b
清肠合剂组	2	8	5	0	0	1.33±0.31^c
透明质酸钠组	1	9	5	0	0	1.37±0.32^c
F						6.143^＊

组别	0	Ⅰ	Ⅱ	Ⅲ	Ⅳ	Nair’s评分/分
假手术组	9	4	2	0	0	1.28±0.25
模型组	0	3	6	5	1	2.27±0.42^b
清肠合剂组	2	8	5	0	0	1.33±0.31^c
透明质酸钠组	1	9	5	0	0	1.37±0.32^c
F						6.143^＊

组别	IL-4	STAT-6	IFN-γ
空白组	1.01±0.16	1.01±0.15	1.00±0.10
假手术组	0.95±0.11	0.84±0.15	1.14±0.20
模型组	0.25±0.06^ab	0.31±0.03^ab	2.06±0.11^ab
清肠合剂组	0.66±0.16^c	0.70±0.09^c	1.35±0.14^c
透明质酸钠组	0.60±0.10^c	0.67±0.04^c	1.48±0.24^c
F	18.486^＊＊	18.388^＊＊	18.522^＊＊