| Objective:Hypertension is a low degree of inflammatory disease,and the immune system is involved in the development of hypertension,and the gap junctions between immune cells are involved in the process of inflammation.The donor of nitric oxide has been widely used in clinic as a blood pressure lowering drug,which has anti-inflammatory effect.The purpose of this study is to investigate whether nitric oxide can modulate the immune inflammatory response through the gap junction between peripheral blood T lymphocytes and ultimately alleviate the target organ damage caused by hypertension.Methods:in vivo experiment:The 12 weeks old of spontaneously hypertensive rats(SHR)and Wistar-Kyoto(WKY)rats as the research object,which divided into WKY group,SHR group and SHR+NO group,8 rats in each group.The SHR+NO intervention group were injected intraperitoneally Sodium nitroprusside according to 10?g/(kg·d)for 4 weeks,then the WKY group and the SHR group were injected intraperitoneally equal amount of saline for 4 weeks.The blood pressure of tail artery in rats was measured by BP-6 noninvasive blood pressure meter;the expression of cell factors(IL-17,IL-2,IL-6,TNF-?and IL-10)in peripheral blood of rats were detected by the method of ELISA;the pathological changes of basilar artery and kidney in rats were observed by HE staining;the T lymphocyte subsets in peripheral blood and the expression of connexin 40(Cx40)in T lymphocytes of rats were detected by flow cytometry.In vitro experiment:the peripheral blood lymphocytes of SHR and WKY rats were extracted under aseptic condition,which were divided into WKY group,SHR group and SHR+NO group,the SHR+NO group was tread with 200?M nitric oxide for 48 hours,the WKY group and SHR group were given equal amount of saline to incubate 48h,after that,the number of cells in the three groups was observed and the expression of Cx40 in the three groups of lymphocytes was detected by the method of western blot and flow cytometry,the expression of IL-6 and TNF-?in cell culture supernatant detected by ELISA.Results:(1)The tail artery systolic pressure of SHR was significantly higher than that of WKY rats(P<0.01,n=8),and the tail artery systolic pressure of SHR+NO group was significantly lower than that of SHR group(P<0.01,n=8).(2)Compared with the WKY group,the expression levels of IL-17,IL-2,IL-6and TNF-?increased in the plasma of SHR group(P<0.01,n=8),and the expression level of IL-10decreased(P<0.01,n=8);the expression levels of IL-17,IL-2 and TNF-?in SHR+NO group were decreased(P<0.01,n=8),and the expression level of IL-6 was decreased(P<0.05,n=8),and the expression level of IL-10 increased(P<0.05,n=8).(3)Compared with the WKY group,the SHR group suffered from the damage of cerebral basilar artery endothelial cells,thickening of blood vessel wall,atrophy of glomerulus and inflammatory cell infiltration,then the inflammatory damage degree of SHR+NO group was lower than that of SHR group.(4)The frequency of CD3~+T cells in SHR group increased comparing with the WKY group(P<0.01,n=8),the frequency of CD3~+T cells in SHR+NO group decreased comparing with the SHR group(P<0.05,n=8).(5)Compared with the WKY group,the frequency of CD4~+T cells and the ratio of CD4~+/CD8~+increased(P<0.01,n=8)and the frequency of CD8~+T cells decreased(P<0.05,n=8)and the frequency of CD4~+CD25~+T cells increased(P<0.01,n=8)in SHR group,then the frequency of CD4~+T cells and the ratio of CD4~+/CD8~+decreased(P<0.05,n=8)and the frequencies of CD8~+T cells and CD4~+CD25~+T cells in SHR+NO group increased(P<0.05,n=8)comparing with the SHR group.(6)The expression frequency of CD4~+Cx40~+in SHR group was significantly higher than that in WKY group(P<0.01,n=8),the expression frequency of CD4~+Cx40~+in SHR+NO group was lower than that in SHR group(P<0.01,n=8);the expression frequency of CD8~+Cx40~+in SHR group was significantly higher than that in WKY group(P<0.05,n=8),the expression frequency of CD8~+Cx40~+in SHR+NO group was lower than that in SHR group(P<0.05,n=8).(7)The expression frequency of Cx40 on T lymphocytes in primary cultured SHR group was significantly higher than that in WKY group(P<0.05,n=6),and the expression frequency of Cx40 on T lymphocyte in SHR+NO group was significantly lower than that in SHR group(P<0.05,n=6).(8)The expression of Cx40 in T lymphocytes in primary cultured in SHR group was significantly higher than that in group WKY(P<0.05,n=3),and the expression of Cx40 in T lymphocyte in SHR+NO group was significantly lower than that in SHR group(P<0.05,n=3).(9)In cell culture supernatant,the expression level of IL-6 and TNF-?in group SHR were higher than those in WKY group(P<0.05,n=6),and the expression level of IL-6 and TNF-?in SHR+NO group were lower than those in SHR group(P<0.05,n=6).Conclusion:The inflammatory damage of cerebral vessels and kidneys occured in SHR,and accompanied with the disorder of peripheral blood T lymphocyte subsets and inflammatory factors;NO can down regulate the expression of Cx40 on peripheral blood T lymphocytes;NO may regulate cytokine release by inhibiting the expression of Cx40 in peripheral blood T lymphocytes,thereby relieving target organ damage caused by hypertension. |
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