Study of ultrasound-guided ropivacaine combined with butorphanol continuous paravertebral block to prevent pain syndrome by evaluating ccl2 gene expression after radical mastectomy

This study aimed to investigate the clinical effect of ultrasound-guided ropivacaine combined with butorphanol continuous paravertebral block in preventing postoperative pain syndrome of breast cancer. For this purpose, 100 women treated for breast cancer from April 2018 to July 2019 were enrolled as research objects. Surgical procedures included local sentinel lymph node biopsy, mastectomy, sentinel lymph node biopsy for mastectomy, modified radical mastectomy, and implantation. The selected patients were randomly divided into two groups: control group (routine operation anesthesia; n = 50) and observation group (ultrasound-guided thoracic paravertebral block before induction of ropivacaine+butorphanol anesthesia; n = 50). The Real-time PCR technique was performed to evaluate CCL2 gene expression. VAS scores were recorded during the postoperative period. Compared with the control group, the observation group had lower VAS scores at six h, 24h, and 48h (P<0.05). The pain effect of the observation group was less than that of the control group. The observation group had better analgesic effects after anesthesia. The observation group had a lower incidence of pain syndrome at the 6th, 8th, and 12th months (P<0.05), and the incidence of pain syndrome in the two groups decreased with the extension of time. The observation group had lower levels of related factors (P<0.05), and the observation group had lower traumatic stress responses. The protein expression of IL-6, IL-17, and CRP in the observation group was lower than that in the control group (P<0.05). The results of CCL2 gene expression also showed that gene expression in the control group increased significantly (P=0.0047). Since the expression of this gene is one of the factors that stimulate pain signals in the body, the method used in the present study was able to reduce the amount of pain significantly. Therefore, the combination of ropivacaine combined with butorphanol ultrasound-assisted paravertebral block can reduce the intensity of postoperative pain in patients with breast cancer surgery, decrease the incidence of pain syndrome, and increase pain tolerance.

Phenylbutyrate facilitates homeostasis of non-resolving inflammatory macrophages

Non-resolving inflammatory monocytes/macrophages are critically involved in the pathogenesis of chronic inflammatory diseases. However, mechanisms of macrophage polarization are not well understood, thus hindering the development of effective strategies to promote inflammation resolution. In this study, we report that macrophages polarized by subclinical super-low dose LPS preferentially expressed pro-inflammatory mediators such as ccl2 (which encodes the protein monocyte chemo attractant protein-1) with reduced expression of anti-inflammatory/homeostatic mediators such as slc40a1 (which encodes the protein ferroportin-1). We observed significantly elevated levels of the autophagy-associated and pro-inflammatory protein p62 in polarized macrophages, closely correlated with the inflammatory activation of ccl2 gene expression. In contrast, we noted a significant increase of ubiquitinated/inactive nuclear-erythroid-related factor 2 (NRF2), consistent with reduced slc40a1 gene expression in polarized macrophages. Addition of the homeostatic restorative agent phenylbutyrate (4-PBA) effectively reduced cellular levels of p62 as well as ccl2 gene induction by super-low dose LPS. On the other hand, application of 4-PBA also blocked the accumulation of ubiquitinated NRF2 and restored anti-inflammatory slc40a1 gene expression in macrophages. Together, our study provides novel insights with regard to macrophage polarization and reveals 4-PBA as a promising molecule in restoring macrophage homeostasis.

5946The immune protein MD-2 is associated with early death in dilated cardiomyopathy and increases M1 macrophage polarization and recruitment in vitro

Objective Dilated cardiomyopathy (DCM) is characterized by systolic dysfunction and dilatation of ventricles. Myocardial inflammation and leukocyte activation and recruitment play a major role in the development and progression of disease. Myeloid differentiation factor-2 (MD-2) is the TLR (toll like receptor)-4 co-receptor and has been shown to be an important risk predictor for mortality of DCM patients. It is expressed in various cell types and mediates TLR-4 dependent inflammation/activation processes. Purpose We examined the impact of MD-2 on mortality of DCM patients and on polarization and recruitment of monocytes in vitro. Methods In 77 DCM patients, divided by median time point of death after first hospital admission into early and late death and alive group, MD-2 was quantified by means of ELISA. In THP-1 monocytes, cytokine secretion was quantified by ELISA after 72h treatment with MD-2 (5μg/mL). Bone marrow derived macrophages (BMDM) were generated from MD-2 KO and WT mice. NFkB phosphorylation (10min) and changes in gene expression (4h) of different adhesion molecules was quantified after treatment with 1 or 10ng/mL LPS. In human umbilical vein endothelial cells (HUVEC), protein kinase B (PKB) phosphorylation was quantified after 15min of treatment with 10ng/mL LPS or 5μg/mL MD-2. CCL2 gene expression in lysed cells (4h) and CCL2 secretion in supernatants (48h) were quantified to. Adhesion of monocytes on treated HUVEC was determined by FACS (Fig.1a). Initial HUVEC treatment with MD-2 (5μg/mL) or LPS (10 or 100ng/mL) took place for 48h. Results We found significant increased MD-2 in early (591.3ng/mL; N=18) vs late death (p=0.015) (369.2ng/mL; N=17) and alive (p≤0.0001) (303.2ng/mL; N=42) patients. Treatment of THP-1 cells (N=5) with MD-2 lead to a significantly increased secretion of inflammatory cytokines IL-8 (p=0.012), IP-10 (p=0.029), and MCP-1 (p=0.032) but not of anti-inflammatory cytokines IL-4, IL-10 and IL-13. Treatment of BMDM obtained from MD-2 KO and WT mice with 10ng/mL LPS lead to a increased phosphorylation of NFkB (N=4; p=0.022) and increased gene expression (N=6) of adhesion molecules VLA-4 (p=0.006) and ICAM-1 (p=0.049) in WT mice but not in KO mice. In HUVEC, LPS (p=0.008) and MD-2 induced a comparable increased phosphorylation of PKB (p=0.008) as well as an increase of CCL2 gene expression (p=0.029) and protein amount (p=0.039). Furthermore, treatment of HUVEC with both MD-2 (p=0.015) and LPS (p=0.0001) lead to a significant increase in monocyte adhesion (Fig.1). Conclusion The impact of MD-2 on cardiac inflammation and macrophage recruitment has not been described yet. In this study, we showed that, in DCM, elevated levels of sMD-2 are associated with early death. Furthermore, we could demonstrate that MD-2 enhances the process of HUVEC based monocyte recruitment. Finally, we could show that MD-2 induces inflammatory monocyte activity and triggers polarization of macrophages towards an inflammatory phenotype.

Аssociation between (rs1024611) -12518A/G CCL2 gene polymorphism and serum level of C-reactive protein with different clinical and serological phenotypes of systemic sclerosis in the Russian cohort of patients

Установлено, что полиморфизмы генов медиаторов воспаления, тканевых матричных белков и ростовых факторов вовлечены в аутоиммунные и фибротические процессы при системной склеродермии (ССД). Хемокин CCL2 является ключевым воспалительным белком, присутствующим в циркулирующей крови и в очагах повреждения кожи больных ССД. Цель настоящего исследования состояла в изучении связи полиморфизма -2518 A/G гена CCL2 с уровнем С-реактивного белка (СРБ) у больных с разными клиническими и серологическими фенотипами ССД. В общей группе пациентов носители генотипа -2518АА имели статистически значимо более высокий средний уровень СРБ по сравнению с носителями аллеля -2518G (AG+GG генотипы) (р=0,032). Сходные различия уровней СРБ были получены у пациентов с диффузной формой ССД и у пациентов с позитивными титрами антител к топоизомеразе I (р=0,041 и р=0,042 соответственно). Выводы. Носительство генотипа -2518AА ассоциировано с высоким уровнем СРБ у больных с плохим прогнозом ССД (диффузная форма и позитивные титры аутоантител к топоизомеразе I) по сравнению с носительством аллеля -2518G. Полиморфизм -2518A/G гена CCL2 может быть использован в качестве генетического маркера тяжести и неблагоприятного прогноза ССД. It has been established that polymorphisms of inflammatory mediator genes, tissue matrix proteins and growth factors are involved in autoimmune and fibrotic processes in systemic scleroderma (SSc). CCL2 chemokine is a key inflammatory protein present in the circulating blood and in the lesions of the skin of patients with SSc. The purpose of this study was to study the association of the polymorphism -2518 A / G of the CCL2 gene with C-reactive protein (CRP) levels among patients with different clinical and serological phenotypes of SJS. In the general group of patients, carriers of the genotype -2518AA had a statistically significant higher average CRP level compared to carriers of the -2518G allele (AG + GG genotypes) (p = 0.032). Similar differences in CRP levels were obtained in patients with a diffuse form of SJS and in patients with positive antibody titers to topoisomerase I (p = 0.041 and p = 0.042, respectively). Conclusions: The carriage of the -2518AA genotype was significantly associated with a high level of CRP in patients with poor prognosis of SSc (presence of a diffuse form and seropositive autoantibody titers to topoisomerase I) compared with patients carrying the -2518G allele. Polymorphism -2518A / G of the CCL2 gene can be used as a genetic marker of severity and poor prognosis for SSc.