Pro-angiogenic effect of SEHM formula on HUVEC cells in vitro and Zebrafish in vivo and action mechanism study / 中国药理学通报

Aim To investigate pro-angiogenic effect of SEHM formula on HUVEC cells in vitro and Zebrafish in vivo and the related action mechanism .Methods VEGFR tyrosine kinase inhibitor II ( VRI)-induced ze-brafish intersegmental vessels ( ISVs ) damaged model was established to observe the protective effect of SE-HM formula on ISVs under fluorescence microscope . The pro-angiogenic effect on subintestinal vessels ( SIVs ) of water decoction of SEHM formula in ze-brafish was observed and the mRNA level of VEGFRs , including flt-1, kdr, kdrl, was measured by real-time PCR.The experimental models of the HUVEC cells were set up and the toxicity and promoting proliferation effect of water decoction of SEHM formula in HUVEC cells were assessed by cell viability assay (MTT), and then the levels of Akt, p38, p-p38, p44/42, p-p44/42, VEGFR-1 were detected by Western blot at 6, 12 and 24 h.Results SEHM formula treatment groups could significantly protect VRI-induced zebrafish ISVs loss(P<0.01) and presented pro-angiogenic effect on zebrafish SIVs obviously(P<0.01).The mRNA level of VEGFRs, including flt-1, kdr, kdrl.was up-regula-ted by SEHM formula treatment group significantly (P<0.05) compared with VRI group.Compared with control group , and SEHM formula treatment group could apparently promote proliferation in HUVEC cells and up-regulate the level of Akt, p38, p-p38, p44/42, p-p44/42, VEGFR-1.Conclusions Water de-coction of SEHM formula could present pro-angiogenic effect on SIVs in zebrafish and promote proliferation of HUVEC cells significantly , and its action mechanism may be related to up-regulating the expression of VEG-FR.

Effects of hepatotrophic factors on the liver after portacaval shunt in rats with portal hypertension / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Portacaval shunt (PCS) prevent hepatotrophic factors from flowing into the liver, but they enter directly the systemic circulation and worsen liver injury. This study was designed to investigate the effects of hepatotrophic factors through the portal vein on the liver in rats with portal hypertension after portacaval shunt.</p><p><b>METHODS</b>Intrahepatic portal hypertension (IHPH) was induced by intragastric administration of carbon tetrachloride, and end-to-side PCS was performed. Eight normal rats served as controls, and eight rats with IHPH served as IHPH model (IHPH group). Another 32 rats with IHPH-PCS were randomly subdivided into 4 groups: normal saline (NS) given to 8 rats, hepatocyte growth factor (HGF) 8, insulin (INS) 8, hepatocyte growth factor and insulin (HGF + INS) 8. Hepatotrophic factors were infused into the portal vein through an intravenous catheter. Portal venous pressure (PVP) was measured. The levels of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were tested biochemically and those of hyaluronic acid (HA) and laminin (LN) were measured by radioimmunoassay. Hepatic fibrosis was assessed histologically and the expression of collagens type I and III were detected immunohistochemically. Ultrastructural change of hepatocytes and the number of mitochondria were observed under an electron microscope. The data were compared between groups and subgroups by Student-Newman-Keuls procedure with SPSS10.0.</p><p><b>RESULTS</b>PVP was significantly higher in the IHPH rats than in the control rats (P < 0.05). The levels of serum ALT, AST, HA, and LN, hepatic fibrosis score, the amount of collagen deposition, collagens type I and III increased more significantly in the IHPH group than in the control rats (P < 0.05). The number of mitochondria decreased more significantly in the IHPH rats than in the control rats (P < 0.05). The levels of serum ALT, AST, HA and LN as well as hepatic fibrosis score, the amount of collagen deposition, and the amount of collagens type I and III in the HGF and HGF + INS rats were significantly lower than those in the NS rats (P < 0.05). The damage to hepatocyte ultrastructure was markedly alleviated and the number of mitochondria was increased more significantly in the HGF and HGF + INS rats than in the NS rats under an electron microscope.</p><p><b>CONCLUSIONS</b>Perfusion of exogenous hepatotrophic factors through the portal vein can alleviate liver injury, minimize the damage to the ultrastructure of hepatocyte, protect liver function, and lessen hepatic fibrosis in rats with portal hypertension after PCS.</p>

Effect of endothelin-1 and its antagonists on the expression of endothelin receptors mRNA in HSC-T6 cells / 中华外科杂志

<p><b>OBJECTIVE</b>To study the effect of endothelin-1 (ET-1) and its antagonists on the expression of endothelin and its receptors mRNA in HSC-T6 cells.</p><p><b>METHODS</b>Cultured HSC-T6 cells were randomly divided into 7 groups: Sham control group, ET-1 group (10 nmol/L ET-1), BQ-123 group [1 micromol/L BQ-123, a selective endothelin receptor A (ETRA) antagonist], BQ-788 group [1 micromol/L BQ-788, a selective endothelin receptor B (ETRB) antagonist], ET-1 + BQ123 group (10 nmol/L ET-1 + 1 micromol/L BQ-123), ET-1 + BQ-788 group (10 nmol/L ET-1 + 1 micromol/L BQ-788) and ET-1 + BQ-788 group (10 nmol/L ET-1 + 1 micromol/L BQ-123 + 1 micromol/L BQ-788). The expression of endothelin receptor mRNA of HSC-T6 cells was determined by reverse-transcription polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>The expression of ETRA mRNA in ET-1 + BQ123 + BQ788 and ET-1 + BQ788 group was significantly lower than ET-1 group (0.329 +/- 0.044 and 0.292 +/- 0.023 vs. 0.440 +/- 0.030 P < 0.05). Compared with ET-1 group, the expression of ETRB mRNA in ET-1 + BQ788 group was down regulated obviously (0.499 +/- 0.136 vs. 0.153 +/- 0.071, P < 0.05). There was no significant difference in ET-1 + BQ123 group and ET-1 + BQ123 + BQ788 group when compared with ET-1 group (0.499 +/- 0.136 vs. 0.496 +/- 0.103 and 0.299 +/- 0.129, P > 0.05).</p><p><b>CONCLUSIONS</b>ET-1 has no obvious effect on the expression of ETRA mRNA in HSC-T6. ET-1 may up-regulate the expression of ETRB mRNA. Act on ETRA receptor, ET-1 can inhibit the expression of ETRB mRNA.</p>