Inhibition of Neutrophil Extracellular Traps Formation by Cl-Amidine Alleviates Lipopolysaccharide-Induced Endometritis and Uterine Tissue Damage

Abstract

Endometritis is a common disease that affects the production in dairy cows and leads to severe losses in the dairy industry. Neutrophil extracellular traps (NETs) formation promotes pathogenic invasions of the lumen of the tissue, leading to inflammatory diseases such as mastitis, pancreatitis, and septic infection. However, research that could show the relationship between NETs and endometritis isscarce. Cl-amidine has been shown to ameliorate the disease squealing and clinical manifestation in various disease models. In this study, we investigated the role of NETs in LPS-triggered endometritis in rats and evaluated the therapeutic efficiency of Cl-amidine. An LPS-induced endometritis model in rats was established and found that the formation of NETs can be detected in the rat’s uterine tissues in vivo. In addition, Cl-amidine treatment can inhibit NETs construction in LPS-induced endometritis in rats. Myeloperoxidase (MPO) activity assay indicated that Cl-amidine treatment remarkably alleviated the inflammatory cell infiltrations and attenuated the damage to the uterine tissue. The Western blot results indicated that Cl-amidine decreased the expression of citrullinated Histone H3 (Cit-H3) and high-mobility group box 1 protein (HMGB1) protein in LPS-induced rat endometritis. The ELISA test indicated that Cl-amidine treatment significantly inhibited the expression of the pro-inflammatory cytokines IL-1β, IL-6, and TNF-α. The NETs were determined by Quant-iTTMPicoGreen dsDNA kit®, which indicated that Cl-amidine significantly inhibited the NETs in rat serum. All results showed that Cl-amidine effectively reduced the expression of Cit-H3 and HMGB1 proteins by inhibiting the formation of NETs, thereby attenuating the inflammatory response to LPS-induced endometritis in rats. Hence, Cl-amidine could be a potential candidate for the treatment of endometritis.

Combination of Doxycycline with Metronidazole Protects against Pyroptosis in Rats with Endometritis through the Modulation of TLR4/NF-κB Pathway

“…Control Injection of 1 mg/kg saline into the uterus Model Injection of 1 mg/kg of 1 mg/mL LPS into the uterus [20] Metronidazole Injection of 1 mg/kg of 1 mg/mL LPS into the uterus, and intraperitoneal injection of 135 mg/kg/day metronidazole for 5 consecutive days [21]…”

“…Grouping of rats and the exact treatments given to every group are presented in Table 1 (Ref. [20][21][22]). After treatments, the remaining total 25 rats (5 rats per group) were euthanized by cervical dislocation after being anesthetized, and uterus was extracted from each rat, fixed for 24 h in 4% paraformaldehyde, dehydrated and embedded in paraffin, and cut into 8-µm-thick sections.…”

Neutrophil extracellular traps primed intercellular communication in cancer progression as a promising therapeutic target

“…Inhibitor of PAD4 Renal IRI Renal IRI mouse [130] Lung injury SAH mouse [131] Hepatocellular carcinoma Hepa1-6 bearing mouse [132] Breast cancer 4T1-bearing mice [28,129] Cl-amidine Inhibitor of PAD4 Mastitis LPS-induced mouse mastitis [133] Endometritis LPS-induced rat endometritis [134] BMS-P5 Inhibitor of PAD4 NETs is crucial for tissue homeostasis and avoiding the presentation of self-antigens [123]. Evidence showed that inhibiting NET formation by the treatment of Dnase I significantly protected hepatocytes and reduced inflammation after liver ischemia/reperfusion (I/R) injury [121], lessened lung injury, and improved survival both in mouse models and in humans with ARDS from pneumonia or sepsis [122].…”

“…It was reported that Cl-amidine could also weaken the inflammatory response of LPS-induced endometritis in rats by decreasing the formation of NETs [134]. BMS-P5 is a novel PAD4-specific inhibitor.…”