ABSTRACT
Objective To investigate the effect of microRNA-22 (miRNA-22) on the expression of P2X7 receptor and inflammatory factors in hippocampus of rats with epilepsy. Methods Healthy SD male rats were intraperitoneal injected with lithium chloride and pilocarpine to induce epilepsy. Three days later, 45 epileptic rats were randomly divided into three groups: epilepsy group( EP group), miRNA-22 agomir group (EF+agomir group) and miRNA-22 agomir control group ( EF+agomir control group). Another 15 healthy rats were selected as control group(N group). The expression of P2X7 protein was detected by West- ern blot and the levels of miRNA-22, P2X7 mRNA, NF-κB mRNA ,IL-1β mRNA were detected by qRT-PCR. Nissl staining was used to observe the damage of Nissl bodies. Results Western blot result showed that compared with the N group(0. 91±0. 10), the level of P2X7 protein in EP group (1. 17±0. 052) in-creased, and the difference was statistically significant (t=-4. 11,P=0. 02). Compared with the EP+ag-omir control group(0. 94± 0. 14),the expression of P2X7 protein in EP+agomir group (0. 66± 0. 06) de-creased and the difference was significant (t=-3. 10,P=0. 04). And the qRT-PCR results showed that compared with N group, the levels of P2X7mRNA (9. 08±0. 94), NF-κB mRNA (20. 10±2. 15) and IL-1β mRNA (50. 64±5. 42) in EP group increased(t=-14. 96,P<0. 05; t=-15. 38,P<0. 05; t=-15. 87,P<0. 05). The expression of P2X7mRNA (1. 31 ± 0. 64), NF-κB mRNA ( 2. 28 ± 1. 10) and IL-1β mRNA (2. 12±1. 20) in EF+agomir group decreased compared with EP group((9. 08± 0. 94),( 20. 10± 2. 15), (50. 64±5. 42)) and EF+agomir control group((7. 03 ±1. 90),(18. 72±1. 76),(47. 39±6. 16)), and the differences were statistically significant(F=29. 77, P<0. 01;F=98. 99, P<0. 05;F=96. 29, P<0. 01). Nissl staining results showed that a large number of morphologically abnormal and disintegrated Nissl bodies could be observed in the hippocampal CA1 and CA3 regions of EP group,which showed a smaller size,irreg-ular morphology,chromatin pyknosis,boundary blur between nucleus and cytoplasm. Compared with the nor-mal group, the difference was significant (P<0. 05). While in miRNA-22 agomir group, the disintegration of Nissl bodies was improved and the number of Nissl bodies increased. Conclusion Intraventricular injec-tion of miRNA-22 agomir can down-regulate the expression of P2X7 receptor and related inflammatory factors in hippocampus of epileptic rats, thus inhibiting seizures.
ABSTRACT
Background:As the endogenous inhibitor of interleukin(IL)-22,IL-22 binding protein(IL-22BP)inhibits the protective effect of IL-22. Expression and significance of IL-22BP in intestinal mucosa of patients with active inflammatory bowel disease(IBD)remain unclear. Aims:To investigate the expression and significance of IL-22BP in intestinal mucosa of patients with active IBD. Methods:A total of 25 Crohn's disease(CD)and 36 ulcerative colitis(UC)patients from Jan. 2017 to Jan. 2018 at Shanghai General Hospital,Shanghai Jiao Tong University School of Medicine were enrolled, and 30 colonic polyp patients were served as controls. The disease activity of CD and UC was assessed. Expressions of IL-22BP mRNA and protein in intestinal mucosa were determined by real-time fluorescent quantitative PCR and immunohistochemistry,respectively. Correlation of IL-22BP protein expression with the disease activity of CD,UC was analyzed. Results:Compared with corresponding control group,expressions of IL-22BP mRNA in intestinal mucosa in CD and UC groups were significantly increased(CD:3.59 ± 0.83 vs. 1.08 ± 0.45,P<0.001;UC:2.19 ± 0.52 vs. 1.05 ± 0.34,P<0.001),and expressions of IL-22BP protein were also significantly increased(CD:6.12 ± 2.30 vs. 1.83 ± 1.86,P<0.001;UC:5.58 ± 2.27 vs. 2.23 ± 1.77,P<0.001). Expression of IL-22BP protein in intestinal mucosa was positively correlated with disease activity of CD(r =0. 649,P <0. 001)and UC(r =0. 732,P <0.001). Conclusions:Expressions of IL-22BP mRNA and protein are increased in intestinal mucosa of patients with active IBD, and the expression of IL-22BP protein is positively correlated with disease activity of IBD.
ABSTRACT
This study was aimed to investigate the effect of baicalin on the proportion of Th22 cells and the concentration of IL-22 bothin vivo andin vitro, in order to explore the immune mechanism of baicalin on inflammatory bowel disease mice model. The 3.5% dextran sodium sulfate (DSS) was used on C57BL/6 mice for the establishment of colitis mice model. Mice were randomly divided into the blank control group, model group, and baicalin group. Flow cytometry and ELISA were used in the detection of the proportion of Th22 cells and concentration of IL-22 in peripheral blood serum, respectively. The spleen lymphocytes of mice were isolated and cultured by baicalin medium (0, 10, 20, 40μM) for 48 h. Flow cytometry was used in the detection of the proportion of Th22 cells. The results showed that baicalin reduced the proportion of Th22 cells and the expression of IL-22 bothin vivo andin vitro experiments. It was concluded that baicalin can inhibit Th22 cell differentiation and expression of IL-22in vitro and DSS-induced colitis mice. It indicated that baicalin had a good treatment potential in Th22 cell-mediated inflammatory diseases.