Hydrogen sulfide protects H9c2 cardiomyocytes against high glucose-induced injury by inhibiting necroptosis

AIM: To study whether hydrogen sulfide(H₂S) protects H9c2 cardiomyocytes against high glucose(HG)-induced injury by inhibiting necroptosis. METHODS: The protein levels of RIP3(an indicator of necroptosis) and cleaved caspase-3 were determined by Western blot. The cell viability was measured by CCK-8 assay. The intracellular le-vels of reactive oxygen species(ROS) were detected by 2', 7'-dichlorfluorescein diacetate staining followed by photofluorography. Mitochondrial membrane potential(MMP) was examined by rhodamine 123 staining followed by photofluorography. The number of apoptotic cells was observed by Hoechst 33258 nuclear staining followed by photofluorography. RESULTS: After the H9c2 cells were treated with HG(35 mmol/L glucose) for 0~24 h, the protein expression of RIP3 in the H9c2 cells was significantly increased at 3 h, 6 h, 9 h, 12 h and 24 h, reaching the maximum level at 24 h. Pretreatment of the cells with 400μmol/L NaHS(a donor of H₂S) or co-treatment of the cells with necrostatin-1(Nec-1; a speci-fic inhibitor of necroptosis) considerably blocked the up-regulation of RIP3 protein induced by HG. Moreover, pretreatment with NaHS or co-treatment with Nec-1 obviously inhibited HG-induced injuries, leading to an increase in the cell viability, and decreases in the generation of ROS and MMP loss. On the other hand, pretreatment with NaHS also reduced the number of apoptotic cells and the protein level of cleaved caspase-3 in the HG-treated H9c2 cardiomyocytes. CONCLUSION: H₂S protects H9c2 cardiomyocytes against HG-induced injury by inhibiting necroptosis.     

Opening of ATP-sensitive K+ channels protects H9c2 cardiac cells against high glucose-induced injury and inflammation by inhibiting TLR4/NF-κB pathway

AIM: To investigate whether the opening of ATP-sensitive K⁺(K_ATP) channels protects H9c2 cardiac cells against high glucose(HG)-induced injury and inflammation by inhibiting the Toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB) pathway. METHODS: The protein levels of TLR4 and NF-κB p65 were determined by Western blot. The levels of interleukin-1β(IL-1β) and tumor necrosis factor-α(TNF-α) were detected by ELISA. The cell viability was measured by CCK-8 assay. Mitochondrial membrane potential(MMP) was examined by rhodamine 123(Rh 123) staining followed by photofluorography. The intracellular levels of reactive oxygen species(ROS) were detected by 2', 7'-dichlorfluorescein- diacetate(DCFH-DA) staining followed by photofluorography. The number of apoptotic cells was observed by Hoechst 33258 nuclear staining followed by photofluorography. RESULTS: After the H9c2 cardiac cells were treated with HG(35 mmol/L glucose) for 24 h, the protein levels of TLR4 and phosphorylated NF-κB p65(p-NF-κB p65) were significantly increased. Pretreatment of the cells with 100 μmol/L diazoxide(DZ, a K_ATP channel opener) for 30 min before exposure to HG considerably blocked the up-regulation of the TLR4 and p-NF-κB protein levels induced by HG. Moreover, co-treatment of the cells with 30 μmol/L TAK-242(an inhibitor of TLR4) obviously inhibited the HG-induced up-regulation of the p-NF-κB p65 protein level. On the other hand, pretreatment of the cells with 100 μmol/L DZ had a clear myocardial protection effect, which attenuated the HG-induced cytotoxicity, inflammatory response, mitochondrial damage, oxidative stress and apoptosis, evidenced by an increase in the cell viability, and decreases in the levels of IL-1β and TNF-α, MMP loss, ROS generation and the number of apoptotic cells. Similarly, co-treatment of H9c2 cardiac cells with 30 μmol/L TAK-242 or 100 μmol/L PDTC(an inhibitor of NF-κB) and HG for 24 h also obviously reduced the above injuries and inflammation induced by HG.CONCLUSION: The opening of K_ATP channels protects H9c2 cardiac cells against HG-induced injury and inflammation by inhibiting the TLR4/NF-κB pathway.     

Angiotensin-(1-7)/Mas receptor axis protects cardiomyocytes against high glucose-induced injury by modulating nuclear factor-κB pathway

AIM: To study whether the angiotensin-(1-7)[Ang-(1-7)]/Mas receptor axis protects cardiomyocytes against high glucose(HG)-induced injury by inhibiting nuclear factor-κB(NF-κB) pathway. METHODS: The cell viability was measured by CCK-8 assay. The intracellular levels of reactive oxygen species(ROS) were detected by DCFH-DA staining. The number of apoptotic cells was tested by Hoechst 33258 nuclear staining. Mitochondrial membrane potential(MMP) was examined by JC-1 staining. The levels of NF-κB p65 subunit and cleaved caspase-3 protein were determined by Western blotting. RESULTS: Treatment of H9c2 cardiac cells with 35 mmol/L glucose(HG) for 30, 60, 90, 120 and 150 min significantly enhanced the levels of phosphorated(p) NF-κB p65, peaking at 60 min. Co-treatment of the cells with 1 μmol/L Ang-(1-7) and HG for 60 min attenuated the up-regulation of p-NF-κB p65 induced by HG. Co-treatment of the cells with Ang-(1-7) at concentrations of 0.1~30 μmol/L and HG for 24 h inhibited HG-induced cytotoxicity, evidenced by an increase in cell viability. On the other hand, 1 μmol/L Ang-(1-7) ameliorated HG-induced apoptosis, oxidative stress and mitochondrial damage, indicated by decreases in the number of apoptotic cells, cleaved caspase-3 level, ROS generation and MMP loss. However, the above cardioprotective effects of Ang-(1-7) were markedly blocked by A-779, an antagonist of Ang-(1-7) receptor(Mas receptor). Similarly, co-treatment of H9c2 cardiac cells with 100 μmol/L PDTC(an inhibitor of NF-κB) and HG for 24 h also obviously reduced the above injuries induced by HG. CONCLUSION: Ang-(1-7)/Mas receptor axis prevents the cardiomyocytes from the HG-induced injury by inhibiting NF-κB pathway.     

Naringin protects against high glucose-induced injury by inhibiting leptin pathway in H9c2 cardiac cells

AIM：To study whether naringin protects H9c2 cardiac cells against high glucose (HG)-induced injury by inhibiting the leptin pathway. METHODS：The expression levels of leptin and leptin receptor (LEPR) were detected by Western blotting. The cell viability was analyzed by CCK-8 assay. The changes of the morphology and the number of apoptotic cells were tested by Hoechst 33258 nuclear staining. The intracellular levels of reactive oxygen species (ROS) were measured by DCFH-DA staining. Mitochondrial membrane potential (MMP) was determined by rhodamine 123 staining. RESULTS：Treatment of the cells with 35 mmol/L glucose (HG) for 6~24 h up-regulated the expression of leptin in H9c2 cardiac cells with the peak value at 9 h. Treatment of the cells with HG for 1~24 h also enhanced the expression of LEPR, peaking at 12 h. Pretreatment with 80 μmol/L naringin for 2 h before exposure of the H9c2 cardiac cells to HG significantly inhibited the up-regulation of both leptin and LEPR induced by HG. Pretreatment of the cells with naringin for 2 h, leptin antagonist for 24 h, or leptin receptor antagonist for 2 h attenuated HG-induced injury in the cardiomyocytes, evidenced by an increase in cell viability, decreases in the number of apoptotic cells and intracellular ROS production as well as a recovery of MMP. CONCLUSION:Naringin may protect the cardiomyocytes against the HG-induced injury by inhibition of the leptin pathway.     

Carnosine protects against high glucose-induced injury in H9c2 cells

AIM： To explore the protective effect of carnosine (Car) on cardiomyocytes with high glucose (HG)-induced injury. METHODS：Rat H9c2 cardiomyocytes were cultured in vitro and divided into three groups: normal control (NC) group, HG group and Car pretreatment (Car+HG) group. The survival rate of H9c2 cells was measured by MTT assay. Intracellular level of reactive oxygen species (ROS) was detected by fluorescent probe DCFH-DA. The protein expression of caspase-8, caspase-9 and caspase-3 was determined by Western blotting. RESULTS：The survival rate of H9c2 cells decreased with the increases in glucose concentration and time, while pretreatment with 20 mmol/L Car could increase the survival rate significantly (P<0.05). The intracellular level of ROS in HG group was significantly increased compared with NC group (P<0.05), while that in Car+HG group was significantly decreased compared with HG group (P<0.05). The expression of caspase-8, caspase-9 and caspase-3 proteins in HG group was significantly increased compared with NC group (P<0.05). Compared with HG group, the expression of caspase-9 and caspase-3 was significantly decreased in Car+HG group (P<0.05), but the expression of caspase-8 did not obviously change (P>0.05). CONCLUSION：Carnosine can protect H9c2 cells against the injury of oxidative stress and apoptosis induced by high glucose.     

Role of ATP-sensitive potassium channels in inhibitory effect of hydrogen sulfide on high glucose-induced inflammation mediated by necroptosis in H9c2 cardiac cells

AIM: To investigate the role of ATP-sensitive potassium (K_ATP) channels in the inhibitory effect of hydrogen sulfide (H₂S) on high glucose(HG)-induced inflammation mediated by necroptosis in H9c2 cardiac cells.METHODS: The expression levels of receptor-interacting protein 3 (RIP3; an indicator of necroptosis) and cyclooxyge-nase-2 (COX-2) were determined by Western blot. The levels of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) were detected by ELISA.RESULTS: After H9c2 cardiac cells were treated with 35 mmol/L glucose (HG) for 24 h, the expression of RIP3 was significantly increased. Pre-treatment of the cells with 100 μmol/L diazoxide (DZ; a K_ATP channel opener) or 400 μmol/L NaHS (a donor of H₂S) for 30 min considerably blocked the up-regulation of RIP3 induced by HG. Moreover, pre-treatment of the cells with 100 μmol/L 5-hydroxydecanoic acid (5-HD; a K_ATP channel blocker) attenuated the inhibitory effect of NaHS on HG-induced up-regulation of RIP3. On the other hand, co-treatment of the cells with 100 μmol/L necrostatin-1 (a specific inhibitor of necroptosis) or pre-treatment of the cells with 100 μmol/L DZ or 400 μmol/L NaHS attenuated HG-induced inflammatory responses, evidenced by decreases in the expression of COX-2 and secretion levels of IL-1β and TNF-α. However, pre-treatment of the cells with 100 μmol/L 5-HD significantly attenuated the above anti-inflammatory effects of NaHS.CONCLUSION: K_ATP channels play an important role in the inhibitory effect of H₂S on HG-induced inflammation mediated by necroptosis in H9c2 cardiac cells.     

Effects of perindopril at different doses on cardiac function and ACE2/Ang-(1-9)/Ang-(1-7) axis of ischemic cardiac dysfunction rabbits

AIM: To investigate the different dose of perindopril on cardiac function in the rabbits with ischemic cardiac dysfunction. METHODS: Male rabbits weighing 2.5~3.0 kg(n=30) were randomly divided into 3 groups(n=10):high dose perindopril group(HD group), low dose perindopril group(LD group) and cardiac dysfunction group(CD group). The Left anterior descending coronary artery of the rabbits was ligatured for model preparation. In HD group, the rabbits were treated with perindopril split normal saline solution(1 g/L)2 mL·kg^-1·d^-1. In LD group, the rabbits were treated with perindopril split normal saline solution(0.33 g/L)2 mL·kg^-1·d^-1. In CD group, the rabbits were treated with normal saline solution 2 mL·kg^-1·d^-1. Four weeks after treatment, the cardiac function was measured via echocardiography, the mRNA expression of angiotensin-converting enzyme 2(ACE2) and angiotensin type 2 receptor(AT2R) was analyzed by real-time PCR, serum angiotensin(Ang)-(1-9) and Ang-(1-7) levels were detected by ELISA. RESULTS: Compared with CD group, the cardiac function of the 2 groups treated with perindopril was significantly improved(P<0.01), and more improvement in HD group was observed than LD group(P<0.05). The serum angiotensin(Ang)-(1-9) and Ang-(1-7) level and the mRNA expression of ACE2 and AT2R in the 2 groups treated with perindopril were significantly improved(P<0.01). Compared with LD group, the mRNA expression of ACE2 and AT2R and the serum levels of Ang-(1-9) in HD group were significant improved(P<0.05), while no difference of serum Ang-(1-7) level was observed. Correlation analysis revealed that the improvement of the cardiac function was associated with serum Ang-(1-9) level, mRNA expression of ACE2 and AT2R(P<0.01), but has no significant correlation with serum Ang-(1-7) level. CONCLUSION: High dose of perindopril may improve more cardiac function in ischemic cardiac dysfunction model in rabbits. The mechanism may relate to increasing serum Ang-(1-7) level to activate AT2R.     

Angiotensin-(1-7) increases expression of ABCA1 and decreases content of cholesterol in THP-1-derived foam cells via AC-cAMP pathway

AIM: To establish the THP-1-derived foam cell formation and to evaluate the effects of angiotensin-(1-7) and MDL (an inhibitor of adenylate cyclase) on the expression of ATP-binding cassete transporter A1(ABCA1) and the content of cholesterol. METHODS: THP-1-derived macrophages were treated with oxidized low-density lipoprotein(ox-LDL) to develop into foam cells. The foam cells were divided into 4 groups: control group, MDL group, Ang-(1-7) group and MDL+Ang-(1-7) group. At 24 h after treatment, the content of cAMP was measured by ELISA. The mRNA and protein levels of ABCA1 were determined by real-time RT-PCR and Western blotting, respectively. The content of cholesterol was detected by high performance liquid chromatography. RESULTS: The cAMP, the mRNA and protein levels of ABCA1 in Ang-(1-7) group were significantly higher, and the content of cholesterol was significantly lower than those in control group (P<0.05). On the contrary, the cAMP, the mRNA and protein levels of ABCA1 in MDL group were significantly lower and the content of cholesterol was significantly higher than those in control group (P<0.05). The results in MDL+Ang-(1-7) group were between Ang-(1-7) group and control group. CONCLUSION: Ang-(1-7) inhibits the formation of foam cells by promoting the expression of ABCA1 and decreasing the content of cholesterol. MDL partly antagonizes the effect of Ang-(1-7) by inhibiting the adenylate cyclase and decreasing the content of cAMP.     

Effect of angiotensin-(1-7) on angiotensin II-induced rat renal interstitial fibroblast activation

AIM: To explore the influence of angiotensin-(1-7) on angiotension II (Ang II)-induced activation and extracellular matrix secretion in rat renal interstitial fibroblasts (NRK-49F cells). METHODS: The NRK-49F cells were maintained and sub-cultured, then the cells were divided into control group, Ang II group, Ang-(1-7) group and Ang II+Ang-(1-7) group. The expression of α-smooth muscle actin(α-SMA),transforming growth factor β₁ (TGF-β₁) and insulin-like growth factor I(IGF-I) was detected by the method of immunocytochemistry when the cells were cultured for 72 h. The content of TGF-β₁, IGF-I and collagen type I(Col I) in the cultured supernatants were measured by ELISA. RESULTS: In control group and Ang-(1-7) group, only basic expression of α-SMA and almost no expression of TGF-β₁, IGF-I and Col I were observed. Compared with control group, the expression of α-SMA, TGF-β₁, IGF-I and Col I was increased in Ang II group. Compared with Ang II group, the expression of α-SMA, TGF-β₁, IGF-I and Col I was significantly decreased in Ang II+Ang-(1-7) group.CONCLUSION: Ang-(1-7) inhibits the activation of renal interstitial fibroblasts and decreases the Ang II induced secretion of Col I by suppressing TGF-β₁ and IGF-I expression.     

Role of ATP-sensitive potassium channels in inhibitory effect of hydrogen sulfide on high glucose-induced injury in H9c2 cardiac cells

AIM: To investigate the roles of ATP-sensitive potassium (K_ATP) channels in high glucose-induced cardiac injury and the inhibitory effect of hydrogen sulfide (H₂S) on the cardiomyocyte injury. METHODS: The expression level of K_ATP channel protein was tested by Western blot. The cell viability was measured by CCK-8 assay. The number of apoptotic cells was observed by Hoechst 33258 nuclear staining. Mitochondrial membrane potential (MMP) was examined by JC-1 staining. RESULTS: After the H9c2 cells were treated with 35 mmol/L glucose (high glucose, HG) for 1~24 h, the protein level of K_ATP channel was significantly reduced at 6 h, 9 h, 12 h and 24 h, reaching the minimum level at 12 h and 24 h. Pretreatment of the cells with 400 μmol/L NaHS (a donor of H₂S) prior to exposure to HG for 12 h considerably blocked the down-regulation of K_ATP channels induced by HG. Pretreatment of the cells with 100 μmol/L mitochondrial K_ATP channel opener diazoxide, 50 μmol/L non-selective K_ATP channel opener pinacidil or NaHS obviously inhibited HG-induced injuries, leading to an increase in the cell viability, and decreases in the number of apoptotic cells and the MMP loss. Pretreatment with 100 μmol/L mitochondrial K_ATP channel antagonist 5-hydroxydecanoic acid or 1 mmol/L non-selective K_ATP channel antagonist glibenclamide attenuated the above cardioprotective effects of NaHS. CONCLUSION: K_ATP channels mediate the inhibitory effect of H₂S on HG-induced cardiac injury.     

Effects of angiotensin-(1-7) on calcification in vascular smooth muscle cells and its signal transduction pathway

AIM: To clarify the effects of angiotensin-(1-7) on the calcification in rat vascular smooth muscle cells(VSMCs) and its signal transduction.METHODS: Calcification of cultured rat VSMCs was prepared by incubation of VSMCs with β-glycerophosphate.Calcification was confirmed by Von Kossa staining.The cells were treated with angiotensin-(1-7).The calcium content,alkaline phosphatases activity,osteocalcin and Cbfa1 mRNA expression were also measured.RESULTS: Angiotensin-(1-7) inhibited the increases of calcium content,alkaline phosphatases activity（P>0.05）,osteocalcin concentration and Cbfa1 mRNA expression in calcified VSMCs（P<0.05）,and the effects of angiotensin II on calcium content,alkaline phosphatases activity,osteocalcin concentration and Cbfa1 mRNA expression in calcified VSMCs were also inhibited （P<0.05）.Angiotensin-(1-7) increased cAMP concentration in calcified VSMCs（P<0.05）and selective PKA inhibitor blocked the effects of angiotensin-(1-7) on calcium content,alkaline phosphatases activity,osteocalcin concentration and Cbfa1 mRNA expression（P<0.05）.CONCLUSION: Angiotensin-(1-7) can inhibit beta-glycerophosphate-induced calcification in VSMCs through cAMP-PKA-Cbfa1 pathway and antagonize the effect of angiotensin II on calcification in VSMCs.     

Ang(1-7) attenuates palmitic acid-induced injury of Min6 cells

AIM: To investigate the effect of angiotensin (1-7)[Ang (1-7)] on palmitic acid (PA)-induced injury of Min6 cells and the potential protective mechanisms of autophagy. METHODS: Cultured Min6 cells were divided into 7 groups:control group, PA group, PA+Ang(1-7) group, PA+Ang(1-7)+A779 group, PA+Ang(1-7)+rapamycin group, Ang(1-7) group and A779 group. The function of Min6 cells was detected by glucose-stimulated insulin release test. Intracellular reactive oxygen species (ROS) production was measured by ROS assay kit. Apoptosis was analyzed by flow cytometry with Annexin V/PI staining. Autophagy-related proteins were determined by Western blot. RESULTS: Compared with control group, the secretion of insulin from Min6 cells in PA group was significantly decreased (P<0.05), and the apoptotic rate was increased (P<0.05). The ratio of LC3-Ⅱ/LC3-I was significantly increased (P<0.05). Compared with PA group, the insulin secretion in PA+Ang(1-7) group was increased (P<0.05). The intracellular ROS level and the A779 and LC3-Ⅱ/LC3-I were significantly decreased (P<0.05). This protective effect of Ang(1-7) was partially blocked by A779 and rapamycin. CONCLUSION: Ang(1-7) attenuates PA-induced Min6 cell injury, and its protective mechanism may be related to inhibiting the activity of autophagy.     

Effects of (+)-2-(1-hydroxyl-4-oxocyclohexyl)ethyl caffeate at different doses on rat arthritis model and whole blood inflammation model

AIM:To evaluate the pharmacodynamics of (+)-2-(1-hydroxyl-4-oxocyclohexyl)ethyl caffeate (HOEC), and to explore the possible causes of non-dose-dependent effects of HOEC on collagen-induced arthritis (CIA) rats using the arachidonic acid (AA) metabolic model in rat whole blood. METHODS:The rat CIA model was used to study the treatment with HOEC at 3 doses. The expression of cytoplasmic phospholipase A₂ (cPLA₂), 5-lipoxygenase (5-LOX) and cyclo-oxygenase-2 (COX-2) was assayed by immunohistochemical method. The effects of HOEC and its in vivo metabolite caffeic acid (CA) on AA metabolite in rat whole blood were measured by ELISA. RESULTS:HOEC had a therapeutic effect on rat CIA, but the curative effect at low dose and middle dose (1 and 3 mg/kg) was better than that at high dose (10 mg/kg). The expression levels of cPLA₂, 5-LOX and COX-2 in joint tissues were decreased. HOEC inhibited the metabolites of LOX and COX pathways in the rat whole blood AA metabolic model, while the inhibitory effect of CA on these metabolites was weaker than that of HOEC. CONCLUSION:The anti-inflammatory effect of HOEC on rat CIA may be associated with the inhibition of cPLA₂, 5-LOX and COX-2 expression in the joint tissues. The non-dose-dependent therapeutic effect of HOEC on rat CIA may due to the weaker inhibitory activity of CA on AA metabolic model in rat whole blood than that of HOEC.     

Effects of FKBP12.6 gene on intracellular Ca2+ concentration in mouse H9c2 (2-1) myocardial cells transfected by ultrasound-mediated destruction of microbubbles

AIM:To investigate the changes of intracellular calcium ion (Ca²⁺) concentration in mouse H9c2 (2-1) cells transfected with or without FK506 binding protein 12.6(FKBP12.6) gene by ultrasound mediated destruction of microbubbles. METHODS:The pcDNA3.1-FKBP12.6 plasmid, mingled with albumin-coated microbubbles agents, was transfected into H9c2 (2-1) cells by ultrasound-mediated destruction of microbubbles. The H9c2 (2-1) cell growth state was investigated by inverted microscope. The changes of intracellular Ca²⁺ concentration was determined by laser scanning confocal microscope. The FKBP12.6 protein expression was checked by immunohistochemistry. RESULTS:As compared with control cells, the H9c2 (2-1) cells, transfected with FKBP12.6 gene, grew better, had higher gross intracellular Ca²⁺ concentration. CONCLUSION:FKBP12.6 gene augments Ca²⁺ concentration in mouse H9c2 (2-1) cells, enhances the contractibility of the myocardial cell, which may be helpful to improve the myocardial dysfunction.     

Exercise training attenuates hypertension progression via activation of central ACE2-Ang(1-7)-Mas axis in prehypertensive rats

AIM: To investigate the effects of exercise training on the progression from prehypertension to hypertension, blood pressure regulation and the angiotensin-converting enzyme 2 (ACE2)-angiotensin (Ang) (1-7)-MAS axis activation in cardiovascular centers, and to elucidate the central mechanisms of exercise training postponing hypertension progression. METHODS: The male spontaneously hypertensive rats (SHR; n=20, 5 weeks old) and normotensive Wistar Kyoto (WKY) rats (n=20) were randomly assigned to sedentary (Sed) group and exercise training (ExT) group. The trained rats run on a treadmill in moderate-intensity for 20 weeks. Systolic blood pressure (SBP) was measured by tail-cuff method. The baroreflex sensitivity (BRS) was assessed by intravenous injection of phenylephrine. The expression of ACE2 and MAS receptor at mRNA and protein levels in baroreflex centers were determined by real-time PCR and Western blot, respectively. Alterations of BRS were evaluated before and after intracerebroventricular injection of MAS receptor agonist Ang (1-7) and its antagonist A779, respectively. RESULTS: Compared with SHR+Sed group, exercise training since prehypertension significantly postponed the development of hypertension, delayed the hypertension progression, and decreased SBP in both SHR and WKY rats (P<0.05). Exercise training enhanced blood pressure regulation and improved the BRS in SHR (P<0.01). The expression of ACE2 and MAS receptor at mRNA and protein levels in the baroreflex centers (rostral ventrolateral medulla, nucleus tract solitarius and paraventricular nucleus) were up-regulated in SHR+ExT group (P<0.05). Central administration of A779 abolished the benefits of exercise-induced improvement of BRS in SHR+ExT group (P<0.01). In contrast, Ang(1-7) improved the BRS in both SHR+Sed group and SHR+ExT group (P<0.05). CONCLUSION: Exercise training postpones hypertension progression and improves blood pressure regulation, which may be associated with the activation of central ACE2-Ang(1-7)-Mas axis.