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Effects of endothelin-1 on hepatic stellate cell proliferation,collagen synthesis and secretion,intracellular free calcium concentration 被引量:14

Effects of endothelin-1 on hepatic stellate cell proliferation,collagen synthesis and secretion,intracellular free calcium concentration
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摘要 AIM: To explore the effects of endothelin-1(ET-1) on hepatic stellate cells (HSCs) DNA uptake, DNA synthesis,collagen synthesis and secretion, inward whole-cell calcium concentration ([Ca^2+]i) as well as the blocking effect of verapamil on ET-l-stimulated release of inward calcium(Ca2+) of HSC in vitro.METHODS: Rat hepatic stellate cells (HSCs) were isolated and cultivated. ^3H-TdR and ^3H-proline incorporation used for testing DNA uptake and synthesis, collagen synthesis and secretion of HSCs cultured in vitro;, Fluorescent calcium indicator Fura-2/AM was used to measure [Ca^2+]i inwardHSCs.RESULTS: ET-1 at the concentration of 5×10^-8 mol/L,caused significant increase both in HSC DNA synthesis (2 247+344 cpm, P<0.05) and DNA uptake (P<0.05) when compared with the control group. ET-1 could also increase collagen synthesis (P<0.05 vs control group) and collagen secretion (P<0.05 vscontrol group). Besides, inward HSC [Ca^2+]i reached a peak concentration (422±98 mol/L, P<0.001)at 2 min and then went down slowly to165+51 mol/L(P<0.01) at 25 min from resting state (39±4 mol/L)after treated with ET-1. Verapamil (5 mol/L) blocked ET-1-activated [Ca^2+li inward HSCs compared with control group(P<0.05). Fura-2/AM loaded HSC was suspended in no Ca^2+ buffer containing 1 mol/L EGTA, 5 rain later, 10^-8 mol/L of ET-1 was added, [Ca^2+]i inward HSCs rose from resting state to peak 399±123 mol/L, then began to come down by the time of 20 min. It could also raise [Ca^2+]i inward HSCs even without Ca^2+ in extracellular fluid, and had a remarkable dose-effect relationship(P<0.05). Meanwhile,verapamil could restrain the action of ET-1(P<0.05).CONCLUSION: Actions of ET-1 on collagen metabolism of HSCs may depend on the transportation of inward wholecell calcium. AIM:To explore the effects of endothelin-1(ET-1) on hepatic stellate cells (HSCs) DNA uptake,DNA synthesis, collagen synthesis and secretion,inward whole-cell calcium concentration ([Ca^(2+)]_i) as well as the blocking effect of verapamil on ET-1-stimulated release of inward calcium (Ca^(2+)) of HSC in vitro. METHODS:Rat hepatic stellate cells (HSCs) were isolated and cultivated.~3H-TdR and ~3H-proline incorporation used for testing DNA uptake and synthesis,collagen synthesis and secretion of HSCs cultured in vitro,Fluorescent calcium indicator Fura-2/AM was used to measure [Ca^(2+)]_i inward HSCs. RESULTS:ET-1 at the concentration of 5×10^(-8) mol/L, caused significant increase both in HSC DNA synthesis (2 247±344 cpm,P<0.05) and DNA uptake (P<0.05) when compared with the control group.ET-1 could also increase collagen synthesis (P<0.05 vs control group) and collagen secretion (P<0.05 vs control group).Besides,inward HSC [Ca^(2+)] _i reached a peak concentration (422±98 mol/L,P<0.001) at 2 rain and then went down slowly to 165±51 mol/L (P<0.01) at 25 min from resting state (39±4 mol/L)after treated with ET-1.Verapamil (5 mol/L) blocked ET-1- activated [Ca^(2+)]_i inward HSCs compared with control group (P<0.05).Fura-2/AM loaded HSC was suspended in no Ca^(2+) buffer containing 1 mol/L EGTA,5 min later,10^(-8) mol/L of ET-1 was added,[Ca^(2+)]_i inward HSCs rose from resting state to peak 399±123 mol/L,then began to come down by the time of 20 min.It could also raise [Ca^(2+)]_i inward HSCs even without Ca^(2+) in extracellular fluid,and had a remarkable dose-effect relationship(P<0.05).Meanwhile, verapamil could restrain the action of ET-1(P<0.05). CONCLUSION:Actions of ET-1 on collagen metabolism of HSCs may depend on the transportation of inward whole- cell calcium.
出处 《World Journal of Gastroenterology》 SCIE CAS CSCD 2004年第18期2697-2700,共4页 世界胃肠病学杂志(英文版)
基金 Supported by the Grant for Nature and Science from Shanghai,No.03ZR 14097
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