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目的:淋巴细胞表达胱硫醚γ裂解酶(cystathionineγ-lyase,CSE)/硫化氢(hydrogen sulfide,H_2S),但其是否参与高血压发病尚不清楚。本课题旨在探讨淋巴细胞CSE/H_2S拮抗高血压的免疫调节机制。方法:收取高血压患者及匹配的健康对照纳入研究。亚甲基蓝法检测外周淋巴细胞H_2S产率,Western blot检测蛋白表达及磷酸化,RT-q PCR检测m RNA表达,biotinswitch法检测蛋白质硫氢化修饰。结果:高血压组外周血淋巴细胞CSE蛋白表达、H_2S产率及IL-10水平明显低于正常血压组,药物治疗血压恢复后CSE蛋白表达、H_2S产率及IL-10水平也恢复至正常水平。SHR大鼠给予Na HS治疗4周后,动脉血压显著下调,同时Th17细胞亚群下调,而Treg亚群上调。分离小鼠脾脏CD~+T细胞,si RNA下调CSE或PAG均可抑制Treg的分化,减少IL-10的分泌。反之CSE过表达或H_2S供体可促进Treg分化和IL-10分泌。提示CD4~+T细胞内源性CSE/H_2S可促进其向Treg亚群分化。Treg细胞的分化受到能量代谢的调节。CSE下调或PAG可抑制AMPK Thr172位点磷酸化,促进m TOR Ser2448位点磷酸化。反之CSE过表达或H_2S供体促进AMPK磷酸化,抑制m TOR磷酸化。AMPK Thr172位点磷酸化受LKB1激酶调控,H_2S可促使LKB1 Cys430位点发生硫氢化修饰进而增加LKB1的磷酸化水平。结论:淋巴细胞内源性H_2S可使LKB1 Cys430位点硫氢化修饰并激活LKB1/AMPK通路,促使Treg细胞的分化,并使Treg募集到肾脏、血管周淋巴节,局部分泌IL-10增加,发挥其抗高血压作用。
AIM: Lymphocytes express cystathionine γ-lyase (CSE) / hydrogen sulfide (H 2 S), but its involvement in the pathogenesis of hypertension remains unclear. This topic aims to investigate the immune regulation mechanism of lymphocyte CSE / H 2 S against hypertension. METHODS: Hypertensive patients and matched healthy controls were included in the study. The production of H 2 S in peripheral lymphocytes was detected by methylene blue method, the protein expression and phosphorylation were detected by Western blot, the expression of m RNA by RT-q PCR and the protein sulfhydryl modification by biotinswitch assay. Results: The expression of CSE, the production of H 2 S and the level of IL-10 in peripheral blood lymphocytes in hypertension group were significantly lower than those in normotensive group. The CSE protein expression, H 2 S production rate and IL-10 level returned to normal after blood pressure recovery . After 4 weeks of NaHS treatment in SHR rats, arterial blood pressure was significantly down-regulated while the Th17 cell subset was down-regulated while the Treg subset was up-regulated. Isolation of murine splenic CD ~ + T cells, si RNA down regulation of CSE or PAG can inhibit Treg differentiation, reduce IL-10 secretion. Conversely, CSE overexpression or H 2 S donor promoted Treg differentiation and IL-10 secretion. It is suggested that endogenous CSE / H 2 S in CD4 ~ + T cells can promote their differentiation to Tregs. Treg cell differentiation is regulated by energy metabolism. Down-regulation of CSE or PAG can inhibit AMPK Thr172 site phosphorylation and promote m TOR Ser2448 site phosphorylation. Conversely, CSE overexpression or H 2 S donor stimulated AMPK phosphorylation and inhibited m TOR phosphorylation. Phosphorylation of AMPK Thr172 site is regulated by LKB1 kinase. H_2S can promote the sulfhydrylation of LKB1 Cys430 site and increase the phosphorylation of LKB1. CONCLUSION: Endogenous H 2 S can lyse LKB1 / AMPK pathway through activating LKB1 / AMPK pathway at LKB1 Cys430 site, promote the differentiation of Treg cells and increase the recruitment of Treg to the kidneys and perivascular lymph nodes with an increase of locally secreted IL-10 Its anti-hypertensive effect.