Effect of Paclitaxel+Hirudin on the TLR4-MyD88 Signaling Pathway During Inflammatory Activation of Human Coronary Artery Smooth Muscle Cells and Mechanistic Analysis

Abstract

Background/Aims: Approximately 10%-20% of patients with acute cardiovascular disease who have received coronary intervention suffer restenosis and high inflammation. The stent compound paclitaxel+hirudin was prepared for the treatment of post-intervention restenosis. This study aimed to explore the anti-inflammatory and anti-restenosis mechanisms of paclitaxel+hirudin with regard to the TLR4/MyD88/NF-κB pathway. Methods: Human coronary artery smooth muscle cells (HCASMCs) at 4-6 generations after in vitro culture were used as a model. Lipopolysaccharide (LPS) was used as an inducer to maximally activate the TLR4/MyD88/NF-κB inflammation pathway. After MyD88 knockdown and selective blocking of MyD88 degradation with epoxomicin, the effects of paclitaxel+hirudin stenting on key sites of the TLR4/MyD88/NF-κB pathway were detected using ELISA, Q-PCR, and western blot analysis. Results: LPS at 1 μg/mL for 48 h was the optimal modeling condition for inflammatory activation of HCASMCs. Paclitaxel+hirudin inhibited the levels of key proteins and the gene expression, except for that of the MyD88 gene, of the TLR4-MyD88 pathway. The trend of the effect of paclitaxel+hirudin on the pathway proteins was similar to that of MyD88 knockdown. After epoxomicin intervention, the inhibitory effects of paclitaxel+hirudin on the key genes and proteins of the TLR4-MyD88 pathway were significantly weakened, which even reached pre-intervention levels. Paclitaxel+hirudin affected the MyD88 protein in a dosage-dependent manner. Conclusion: The paclitaxel+hirudin compound promotes MyD88 degradation in the TLR4/MyD88/NF-κB pathway to reduce the activity of TLR4 and NF-κB p65 and to weaken the LPS-initiated inflammatory reactions of IL-1β, IL-6, and TNF-α.

Effect of infection after liver cancer interventional therapy on T lymphocyte subsets and Toll-like receptors in peripheral blood mononuclear cells and its mechanism

“…22 There is also evidence suggesting that TLR2 mainly recognizes Gram-positive bacteria while TLR4 -Gram-negative bacteria. 23 Our study demonstrated that the expression levels of TLR2 and TLR4 in the non-infection group decreased after surgery, while the levels in the infection group increased compared to those before surgery. These results suggest that surgical treatment can improve TLR2 and TLR4 levels in patients.…”

Hirudin Regulates Vascular Function in Chronic Renal Failure through Modulating Macrophage Polarization

“…Han et al have uncovered that Hiru attenuated kidney damage by suppressing inflammation in diabetic nephropathy [ 15 ]. Furthermore, Li et al have illuminated that paclitaxel combined with Hiru eases inflammatory reactions of human coronary artery smooth muscle cells [ 16 ]. What is more, Hiru partially reverses the downregulated Arg-1 expression in streptozotocin-induced diabetes [ 17 ].…”

“…Also, more attention has been paid on the anti-inflammatory effects of Hiru. A large number of researches have mentioned that Hiru decreases inflammatory response in renal interstitial fibrosis [ 14 ], diabetic nephropathy [ 15 ], and acute cardiovascular disease [ 16 ]. Meanwhile, Pang et al have uncovered that Hiru could elevate the expression of arginase-1 (Arg-1), a functional marker of M2 phenotype in high glucose-induced diabetes [ 17 ].…”

Tanshinone IIA reduces pyroptosis in rats with coronary microembolization by inhibiting the TLR4/MyD88/NF-κB/NLRP3 pathway

“…TLRs are widely distributed, which can directly recognize the highly conserved specific molecular structure shared by certain pathogens or their products, and trigger a series of signal transductions [ 23 , 24 ]. TLR4 is a key upstream immune response regulatory factor.…”