A newly improved method of primary cell culture: Tissue block with continuous adhesion subculture in skin fibroblast.

BACKGROUND: Fibroblasts (FBs) have been widely used as a typical in vitro cell model for investigating the biological processes and cell pathophysiological mechanisms. However, FBs are prone to senescence in cell culture process after several passages. Thus, a new approach to cell culture is quite required to enhance the viability of cells. OBJECTIVE: To explore a novel method of cell culture based on skin FBs. METHODS: Dermal tissue blocks were obtained from BALB/c neonatal mice and randomly divided into experimental group and control group. The experimental group received the newly improved culture method, namely, continuous adherence subculture of tissue block (CASTB) method; while the traditional subculture method was applied in the control group. Cells at 1st, 5th and 10th passages were collected and identified by using histological/immunohistochemical and western blot analysis. Cellular viability, proliferation, senescence and apoptosis were analyzed through application of cell growth curve, CCK-8 assay, Ki67 assay, PCNA protein analysis, ß-galactosidase staining, flow cytometry and western blot analysis. RESULTS: Cells under two culture patterns exhibited spindle/irregular shape and vimentin positive expression. With the increase of passage times, the cellular growth rate in the control group gradually decreased, but no alterations emerged from the experimental group. CASTB method remarkably promoted cell growth and proliferation. Moreover, a greatly lower apoptosis and senescence tendency appeared in the experimental group than the control group with passages increasing. CONCLUSION: The method of CASTB is superior to traditional subculture, offering a large number of primary FBs with higher efficiency and success rate and being worth of further popularization and application.

Proanthocyanidins: A novel approach to Henoch­Schonlein purpura through balancing immunity and arresting oxidative stress via TLR4/MyD88/NF­κB signaling pathway (Review).

Henoch-Schonlein purpura (HSP), a recurrent and immunoglobulin (Ig)A-mediated vasculitis, presents not only as skin lesions but also as systemic involvement that can be life-threatening. Although the etiology of HSP remains unknown, immune imbalance and oxidative stress (OS) are primary contributors to its pathogenesis, alongside the abnormal activation of Toll-like receptor (TLR)/myeloid differentiation primary response gene 88 (MyD88)/nuclear factor-κB (NF-κB) pathway. TLRs, especially TLR4, stimulate downstream signaling molecules such as NF-κB and proinflammatory cytokines, which are released when TLRs combine with the key adapter molecule MyD88. This leads to the activation of T helper (Th) cell 2/Th17 and overproduction of reactive oxygen species (ROS). The function of regulatory T (Treg) cells is suppressed in the process. Th17/Treg imbalance then produces various inflammatory cytokines to promote proliferation and differentiation of B cells and the secretion of antibodies. IgA is secreted, and it binds to vascular endothelial surface receptors where the complex induces injury of the vascular endothelial cells. Additionally, excessive ROS creates OS that leads to an inflammatory response and vascular cell apoptosis or necrosis, thereby contributing to vascular endothelial damage and HSP occurrence. Proanthocyanidins are active compounds naturally enriched in fruits, vegetables and plants. Proanthocyanidins have diverse properties, including anti-inflammatory, antioxidant, antibacterial, immunoregulatory, anticarcinogenic and vascular protective effects. Proanthocyanidins are used in the management of various diseases. Proanthocyanidins regulate T cells, equilibrate immunity and arrest OS by inhibiting the TLR4/MyD88/NF-κB signaling pathway. Considering the pathogenesis of HSP and the properties of proanthocyanidins, the present study hypothesized that these compounds may potentially lead to HSP recovery through modulating the immune equilibrium and preventing OS by inhibiting the TLR4/MyD88/NF-κB pathway. To the best of our knowledge, however, little is known about the positive effects of proanthocyanidins against HSP. The present review summarizes the potential of proanthocyanidins to treat HSP.

Natural Plant Extract - Loganin: A Hypothesis for Psoriasis Treatment Through Inhibiting Oxidative Stress and Equilibrating Immunity via Regulation of Macrophage Polarization.

Psoriasis, a chronic immune-mediated inflammatory skin disease, influences approximately 2-3% of the world's population. At present, the etiology of psoriasis remains unclear and there is still no causal treatment available. Recent studies indicate that oxidative stress (OS) and T cells dysregulation may participate in the pathogenesis of psoriasis, among which M1-dominant macrophage polarization is a crucial contributor. Macrophages mainly polarize into two different subsets, ie, classically activated macrophage (M1) and alternatively activated macrophage (M2). M1 polarization tends to exacerbate psoriasis via producing substantial reactive oxygen species (ROS) and inflammatory mediators, to encourage OS invasion and T cells dysregulation. Thus, targeting M1 polarization can be a possible therapeutic alternative for psoriasis. Loganin, belonging to iridoid glycosides, is a pharmaceutically active ingredient originated from Cornus officinalis, exerting multiple biological activities, eg, immunomodulation, antioxidation, anti-inflammation, etc. More importantly, it could effectively suppress M1 polarization, thereby arresting OS aggression and T cells' dysregulation. Numerous studies have confirmed that loganin is quite reliable for diseases treatment via suppressing M1 polarization. Nevertheless, reports about loganin treating psoriasis have seldom appeared so far. Accordingly, we hold a hypothesis that loganin would availably manage psoriasis through preventing M1 polarization. Data from previous studies guarantee the potential of loganin in control of psoriasis.