β-estradiol alleviates hypertension- and concanavalin A-mediated inflammatory responses via modulation of connexins in peripheral blood lymphocytes

Gap junctions (GJs) formed by connexins (Cxs) in T lymphocytes have been reported to have important roles in the T lymphocyte-driven inflammatory response and hypertension-mediated inflammation. Estrogen has a protective effect on cardiovascular diseases, including hypertension and it attenuates excessive inflammatory responses in certain autoimmune diseases. However, the mechanisms involved in regulating the pro-inflammatory response are complex and poorly understood. The current study investigated whether β-estradiol suppresses hypertension and pro-inflammatory stimuli-mediated inflammatory responses by regulating Cxs and Cx-mediated GJs in peripheral blood lymphocytes. Male, 16-week-old spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY) rats were randomly divided into the following three groups: WKY rats, vehicle (saline)-treated SHRs, and β-estradiol (20 µg/kg/day)-treated SHRs. β-estradiol was administered subcutaneously for 5 weeks. Hematoxylin and eosin staining was performed to evaluate target organ injury. Flow cytometry and ELISA were used to measure the populations of T lymphocyte subtypes in the peripheral blood, and expression of Cx40/Cx43 in T cell subtypes, and pro-inflammation cytokines levels, respectively. ELISA, a dye transfer technique, immunofluorescence and immunoblotting were used to analyze the effect of β-estradiol on pro-inflammatory cytokine secretion, Cx-mediated GJs and the expression of Cxs in concanavalin A (Con A)-stimulated peripheral blood lymphocytes isolated from WKY rat. β-estradiol significantly decreased blood pressure and inhibited hypertension-induced target organ injury in SHRs. Additionally, β-estradiol treatment significantly improved the immune homeostasis of SHRs, as demonstrated by the decreased percentage of cluster of differentiation (CD)4(+)/CD8(+) T-cell subset ratio, reduced serum levels of pro-inflammatory cytokines and increased the percentage of CD4(+)CD25(+) T cells. β-estradiol also markedly reduced the expression of Cx40/Cx43 in T lymphocytes from SHRs. In vitro, β-estradiol significantly suppressed the production of pro-inflammatory cytokines, reduced communication via Cx-mediated gap junctions and decreased the expression of Cx40/Cx43 in Con A-stimulated lymphocytes. These results indicate that β-estradiol attenuates inflammation and end organ damage in hypertension, which may be partially mediated via downregulated expression of Cxs and reduced function of Cx-mediated GJ.