Tongxinluo Reverses the Hypoxia-suppressed Claudin-9 in Cardiac Microvascular Endothelial Cells / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Claudin-5, claudin-9, and claudin-11 are expressed in endothelial cells to constitute tight junctions, and their deficiency may lead to hyperpermeability, which is the initiating process and pathological basis of cardiovascular disease. Although tongxinluo (TXL) has satisfactory antianginal effects, whether and how it modulates claudin-5, claudin-9, and claudin-11 in hypoxia-stimulated human cardiac microvascular endothelial cells (HCMECs) have not been reported.</p><p><b>METHODS</b>In this study, HCMECs were stimulated with CoCl2to mimic hypoxia and treated with TXL. First, the messenger RNA (mRNA) expression of claudin-5, claudin-9, and claudin-11 was confirmed. Then, the protein content and distribution of claudin-9, as well as cell morphological changes were evaluated after TXL treatment. Furthermore, the distribution and content histone H3K9 acetylation (H3K9ac) in the claudin-9 gene promoter, which guarantees transcriptional activation, were examined to explore the underlying mechanism, by which TXL up-regulates claudin-9 in hypoxia-stimulated HCMECs.</p><p><b>RESULTS</b>We found that hypoxia-suppressed claudin-9 gene expression in HCMECs (F = 7.244; P = 0.011) and the hypoxia-suppressed claudin-9 could be reversed by TXL (F = 61.911; P = 0.000), which was verified by its protein content changes (F = 29.142; P = 0.000). Moreover, high-dose TXL promoted the cytomembrane localization of claudin-9 in hypoxia-stimulated HCMECs, with attenuation of cell injury. Furthermore, high-dose TXL elevated the hypoxia-inhibited H3K9ac in the claudin-9 gene promoter (F = 37.766; P = 0.000), activating claudin-9 transcription.</p><p><b>CONCLUSIONS</b>The results manifested that TXL reversed the hypoxia-suppressed claudin-9 by elevating H3K9ac in its gene promoter, playing protective roles in HCMECs.</p>

Tongxinluo inhibits cyclooxygenase-2, inducible nitric oxide synthase, hypoxia-inducible factor-2α/vascular endothelial growth factor to antagonize injury in hypoxia-stimulated cardiac microvascular endothelial cells / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Endothelial dysfunction is considered as the initiating process and pathological basis of cardiovascular disease. Cyclooxygenase-2 (COX-2) and prostacyclin synthase (PGIS), inducible nitric oxide synthase (iNOS) and endothelial NOS (eNOS) are key enzymes with opposing actions in inflammation and oxidative stress, which are believed to be the major driver of endothelial dysfunction. And in hypoxia (Hx), Hx-inducible factor (HIF)-1α and HIF-2α are predominantly induced to activate vascular endothelial growth factor (VEGF), resulting in abnormal proliferation. Whether and how Tongxinluo (TXL) modulates COX-2, PGIS, iNOS, eNOS, HIF-1α, HIF-2α, and VEGF in Hx-stimulated human cardiac microvascular endothelial cells (HCMECs) have not been clarified.</p><p><b>METHODS</b>HCMEC were treated with CoCl 2 to mimic Hx and the mRNA expressions of COX-2, PGIS, iNOS, eNOS, HIF-1α, HIF-2α, and VEGF were first confirmed, and then their mRNA expression and protein content as well as the cell pathological alterations were evaluated for TXL treatment with different concentrations. In addition, the effector molecular of inflammation prostaglandin E 2 (PGE 2 ) and the oxidative marker nitrotyrosine (NT) was adopted to reflect HCMEC injury.</p><p><b>RESULTS</b>Hx could induce time-dependent increase of COX-2, iNOS, HIF-2α, and VEGF in HCMEC. Based on the Hx-induced increase, TXL could mainly decrease COX-2, iNOS, HIF-2α, and VEGF in a concentration-dependent manner, with limited effect on the increase of PGIS and eNOS. Their protein contents verified the mRNA expression changes, which was consistent with the cell morphological alterations. Furthermore, high dose TXL could inhibit the Hx-induced increase of PGE 2 and NT contents, attenuating the inflammatory and oxidative injury.</p><p><b>CONCLUSIONS</b>TXL could inhibit inflammation-related COX-2, oxidative stress-related iNOS, and HIF-2α/VEGF to antagonize Hx-induced HCMEC injury.</p>

Protective effect of nourishingyin and promoting blood flow recipe on kidney of diabetic rat / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the protective effects of Nourishingyin and Promotingblood flow recipe (NYPBR) on the kidney of diabetic rat.</p><p><b>METHOD</b>SD rats were divided into 3 groups at random: control group, diabetes group and NYPBR group. The latter two groups were injected intraperitoneally with streptozotocin to induce diabetes model. Rats in NYPBR group were fed NYPBR solution (3 g x d(-1)), with dose equivalent to the clinical use in the patients. Rats in the other groups were fed equivalent water. 10 weeks after diabetes was induced, the inducible nitric oxide synthase (iNOS) mRNA expression in the renal cortex was detected by RT-PCR, and its protein content by Western blotting. Immunohistochemistry was used to detect the formation of nitrotyrosine (NT), a specific marker of peroxynitrite (ONOO-). The morphological changes of renal cortex were observed under optical microscope. Superoxide dismutase (SOD) and malondialdehyde (MDA) in renal cortex, blood glucose, 24 h urine protein content and creatinine clearance rate in different groups were detected.</p><p><b>RESULT</b>Compared with control group, the iNOS mRNA expression (0.90 +/- 0.10) and its protein content (43.00 +/- 6.08), and NT content (87.23 +/- 5.94) increased significantly in diabetes group, in accord with the pathological changes of renal cortex and renal dysfunction. NYPBR can attenuate the pathological alterations.</p><p><b>CONCLUSION</b>NYPBR could decrease iNOS mRNA expression and its protein content, and reducing the overformation of ONOO-, thus protecting the kidney of diabetic rat from injury.</p>

Effect of Tongxinluo on 7 gene expression profile associated with vascular endothelium injure of rats with deficiency of vital energy or qi stagnation / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate 7 gene expression profile associated with inflammation and oxidative stress in vascular endothelium injure of rats with deficiency of vital energy or qi stagnation, and the effect of Tongxinluo on gene expression profile.</p><p><b>METHOD</b>The model of vascular endothelium injury of rats with deficiency of vital energy or qi stagnation were established by using high L-methionine, with load-carrying swimming or being fastened, respectively. RT-PCR and SAGE database which is available in NCBI, were used to analyze the changes of 7 gene expression related with inflammation and oxidative stress in endothelium injure and the effect of Tongxinluo on the gene expression profile.</p><p><b>RESULT</b>Compared with control group, the gene expression of inflammation related COX-1, COX-2, oxidative stress related iNOS, SOD and blood vessel vasomotion related eNOS, ECE, increased in deficiency of vital energy group (P < 0.05 or P < 0.01), and the gene expression decreased with Tongxinluo treatment (P < 0.05 or P < 0.01). The gene expression of COX-1, COX-2, iNOS and eNOS, ECE, increased (P < 0.01), but the gene expression of PCS and SOD decreased (P < 0.01), in qi stagnation group, and the disorder of gene expression improved with treatment of Tongxinluo (P < 0.01).</p><p><b>CONCLUSION</b>The 7 gene expression related to vascular endothelium injure were not the same in rat with deficiency of vital energy or qi stagnation, and Tongxinluo could regulate the disorder of the gene expression, protecting vascular endothelium from injure.</p>