Regulation of Tripterygium wilfordii Polycoride Tablet towards miR-146a, miR-146b, and TLR4/MyD88 dependent signaling pathway in ulcerative colitis rat model
10.7501/j.issn.0253-2670.2016.10.016
- Author:
Dan-Ping QIN
1
Author Information
1. Department of Digestion, First Affiliated Hospital of Zhejiang Chinese Medical University
- Publication Type:Journal Article
- Keywords:
MiR-146a;
MiR-146b;
TLR4/MyD88 dependent signaling pathway;
Tripterygium wilfordii Polycoride Tablet;
Ulcerative colitis
- From:
Chinese Traditional and Herbal Drugs
2016;47(10):1723-1730
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To study the regulatory effect of Tripterygium wilfordii Polycoride Tadlet (TWPT) towards miR-146a, miR-146b, and TLR4/MyD88 dependent signaling pathway in TNBS/ethanol ulcerative colitis (UC) rat model. Methods: TNBS enema was adopted to build TNBS/ethanol UC rat model. After the modeling procedure, 90 male Wistar rats were divided into six groups, including normal, model, low-, mid-, high-dose TWPT, and azathioprine (AZA) groups, and each for 15 rats. All rats in each group were administered with corresponding medicines for 14 d. After 14 d administration, corresponding colon tissues were taken to undergo general and microscopic evaluation. qPCR was adopted to test the expression of miR-146a and miR-146b. Western blotting analysis and RT-PCR were adopted to test the mRNA and protein expression levels of TLR4/MyD88 dependent signaling pathway related molecular, including TLR4, MyD88, TRAF-6, NF-κB, TNF-α, and IL-1β. Results: DAI, general and microscopic evaluation all showed that TNBS/ethanol UC rat model was successfully established. TWPT could improve UC-related clinical manifestation and promote the colonic mucosa healing procedure and such effect was equal to AZA. qRT-PCR showed that the expression of miR-146a and miR-146b in model group was significantly superior to that in normal group (P < 0.01). Compared with the model group, TWPT and AZA could significantly inhibit the expression of miR-146a and miR-146b (P < 0.01). The mid-dose TWPT showed the strongest inhibitory effect. RT-PCR and Western blotting results showed that the expression of TLR4/MyD88 dependent signaling pathway related molecular in model group was significantly superior to that in normal group either in mRNA or protein levels (P < 0.01). Compared with model group, TWPT could inhibit the expression of each spot in TLR4/MyD88 dependent signaling pathway in a dose-dependent manner. The inhibitory effect of high-dose TWPT towards the above molecular was superior to that in model group either in mRNA or protein levels (P < 0.05). The inhibitory effect of high-dose TWPT towards upstream molecular of TLR4/MyD88 dependent signaling pathway (TLR4/MyD88/TRAF-6/NF-κB) was slightly superior to that in AZA group either in mRNA or protein levels. However, such inhibitory effect towards terminal inflammatory cytokines (TNF-α and IL-1β) was slightly inferior to that in AZA group either in mRNA or protein levels. All the above differences had no statistical significance (P > 0.05). Conclusion: In TNBS/ethanol UC rat model, TWPT could inhibit the expression of miR-146a, miR-146b, and TLR4/MyD88 dependent signaling pathway. The inhibitory effect of TWPT towards pathway and inflammatory cytokines shows a dose-dependent manner.
