Paeoniflorin suppresses kidney inflammation by regulating macrophage polarization via KLF4-mediated mitophagy.

BACKGROUND: Macrophages M1 polarization involved in the process of renal inflammatory injury, is a well-established hallmark of chronic kidney disease (CKD). Paeoniflorin (PF), a water-soluble monoterpene glycoside extracted from Paeonia lactiflora, revealed renal anti-inflammatory activities in our previous study. However, the potential molecular mechanism of PF on CKD remains unknown. PURPOSE: The present study aims to investigate the regulation of PF on macrophage polarization in CKD. METHODS: A CKD model was established by cationic bovine serum albumin and a murine macrophage cell line RAW264.7 induced with lipopolysaccharide (LPS) were used to clarify the underlying mechanisms of PF in CKD. RESULTS: Results showed that PF exhibited favorable protective effects on CKD model mice by promoting renal function, ameliorating renal pathological injury and podocyte damage. Furthermore, PF inhibited the infiltration of M1 macrophage marker CD68 and iNOS in kidney tissue, but increased the proportion of M2 macrophage marker CD206. In RAW264.7 cells stimulated with LPS, the levels of cytokines including IL-6, IL-1ß, TNF-α, MCP-1 were lessened under PF treatment, while the levels of Arg1, Fizz1, IL-10 and Ym-1 were augmented. These results indicated that PF promoted macrophage polarization from M1 to M2 in vivo and in vitro. More importantly, PF repaired the damaged mitochondria through increasing mitochondrial membrane potential and reducing ROS accumulation. The mitophagy-related proteins PINK1, Parkin, Bnip3, P62 and LC3 were up-regulated by PF, accompanied by the incremental expressions of Krüppel-like transcription factor 4 (KLF4). Moreover, the promotion of mitophagy and inhibition of M1 macrophage polarization owing to PF were reversed by mitophagy inhibitor Mdivi-1 or silencing KLF4. CONCLUSION: Overall, PF suppressed renal inflammation by promoting macrophage polarization from M1 to M2 and inducing mitophagy via regulating KLF4. It is expected to provide a new strategy for exploring the effects of PF in treating CKD.

An NK cell line (NK92-41BB) expressing high levels of granzyme is engineered to express the high affinity chimeric genes CD16/CAR.

Natural killer (NK) cells are known to play a role in mediating innate immunity and have been implicated in mediating anti-tumor responses via antibody-dependent cell-mediated cytotoxicity (ADCC) based on the reactivity of CD16 with the Fc region of human IgG1 antibodies. The NK-92 cell line, devoid of CD16 and derived from a lymphoma patient, has been well characterized. The adoptive transfer of irradiated NK-92 cells demonstrated safety and showed preliminary evidence of clinical benefit for cancer patients. The molecules 41BB and CD3 are commonly used as stimulators in the CAR structure, and their expression in NK cells can promote the activation of NK cells, leading to the enhanced perforin- and granzyme-mediated lysis of tumor cells. This study showed that genetically modified NK-92 cells combined with antibody-mediated ADCC using rituximab and trastuzumab monoclonal antibodies lysed tumor cells more efficient than the NK-92 cell lines. It also showed that the anti-tumor activity of chimeric stimulator molecules of the CAR-modified CD16 receptor was stronger than that of CD16 (allotype V158). These studies provide a rationale for the use of genetically modified NK-92 cells in combination with IgG1 anti-tumor monoclonal antibodies. We also provide a rationale for the chimeric modified CD16 receptor that can improve the anti-tumor effect of NK92 cells via ADCC. SUPPLEMENTARY INFORMATION: The online version of this article (10.1007/s10616-021-00476-1) contains supplementary material, which is available to authorized users.

Role of anti-TNF-α therapy in fat graft preservation.

BACKGROUND/OBJECTIVE: In this study, we evaluated the role of antitumor necrosis factor-alpha (TNF-α) therapy in the decrease of adipocyte apoptosis and weight preservation of fat grafts in the rat model. METHODS: A total of 64 rats were randomly divided into 2 groups, with 32 rats in each group. Autologous fat tissue was grafted subcutaneously on the back of each rat. For the experimental group, antirat TNF-α monoclonal antibody was injected into the fat grafts during operation. No treatment was given to the tissue in the control group. Eight rats in each group were killed respectively, at days 7, 14, 30, and 60 postoperatively and sampled for assessments of weight preservation, gene expression of TNF-α, histology, and adipocyte apoptosis. RESULTS: There were no significant differences in the weight of fat tissues between the control group and the experimental group at days 7, 14, and 30 postoperatively (P > 0.05). However, the preservation ratio of the tissue was 65.36% ± 14.98% in the antirat TNF-α antibody-treated group when compared with the weight at transplantation, which was significantly higher than the control group (44.63% ± 10.39%) 60 days after the operation (P < 0.05). The numbers of apoptotic cells in the control group were 15.6 ± 3.17, 24.6 ± 4.34, 22.8 ± 2.42, and 27 ± 3.83 per field at different postoperative intervals. However, the numbers of apoptotic cells in the tissues treated with TNF-α antibody were significantly lower than that in the control group, which was 1 ± 0.63, 4 ± 1.41, 6 ± 2.08, and 7.2 ± 2.82 per field (P < 0.05). Gene expression showed that the expression of TNF-α was lower in the experimental group than the control group at days 7 and 14 postoperatively (P < 0.05). CONCLUSION: The results indicate that antirat TNF-α monoclonal antibody can preserve the quality of the transplanted fat tissue.