AIM:To study the effect of copper transporting P-typeATPase in copper metabolism of hepstocyte andpathogenesis of Wilson disease(WD).METHODS:WD copper transporting properties in someorganelles of the cultured hepstocytes were studied fromWD patients and normal controls.These culturedhepstocytes were incubated in the media of copper 15mg·L~(-1) only,copper 15 mg·L~(-1) with vincristine(agonist of P-type ATPase)0.5mg·L~(-1),or copper 15 mg·L~(-1) withvanadate(antagonist of P-type ATPase)18 39mg.L~(-1)separately.Mlcrosome(endoplasmic reticulum and Golgiapparatus),lysosome,mitochondria,and cytosol wereisolated by differential centrifugstion.Copper contents inthese organelles were measured with atomic absorptionspectrophotometer,and the influence in copper transportionof these organelles by vanadate and vincristine werecomparatively analyzed between WD patients and controls.WD copper transporting P-type ATPase was detected bySDS-PAGE in conjunction with Western blot in liver samplesof WD patients and controls.RESULTS:The specific WD proteins(Mr155 000 lanes)wereexpressed in human hepatocytes,including the control andWD patients.After incubation with medium containingcopper for 2 h or 24 h,the microsome copper concentrationIn WD patients was obviously lower than that of controls,and the addtion of vanadste or vincristine would change thecopper transporting of microsomes obviously.Whenincubated with vincristine,levels of copper in microsomewere significantly increased,while incubated with vanadate,the copper corcentrations in microsome were obviouslydecreased.The results indicated that there were WDproteins,the copper tmnsportion P-type ATPase in themicrosome of hepstocytes.WD patients possessedabnormal copper transporting function of WD protein in themicrosome,and the agonist might correct the defect of copper transportion by promoting the activity of coppertransportion P-type ATPase.CONCLUSION:Copper tmnsportion P-type ATPase plays animportant role in hepatocytic copper metabolism.Dysfunction of hepatocytic WD protein copper transportionmight be one of the most important factors for WD. AIM: To study the effect of copper transporting P-type ATPase in copper metabolism of hepstocyte and pathogenesis of Wilson disease (WD). METHODS: WD copper transporting properties in someorganelles of the cultured hepatic cells were studied from WD patients and normal controls. The cultured hepatocytes were incubated in the media of copper 15 mg · L -1 only, copper 15 mg · L -1 with vincristine (agonist of P-type ATPase) 0.5 mg · L -1 or copper 15 mg · L -1 (-1) with vanadate (antagonist of P-type ATPase) 18 39 mg.L ~ (-1) separately.Microsome (endoplasmic reticulum and Golgiapparatus), lysosome, mitochondria, and cytosol wereisolated by differential centrifugation.Copper contents inthese organelles were measured with atomic absorption spectrophotometer, and the influence in copper transportion of these organelles by vanadate and vincristine werecomparatively analyzed between WD patients and controls. WD copper transporting P-type ATPase was detected by SDS-PAGE in conjunction with Western blot in liver samples of W D patients and controls .RESULTS: The specific WD proteins (Mr155 ​​000 lanes) wereexpressed in human hepatocytes, including the control and WD patients. After incubation with medium containing cops for 2 h or 24 h, the microsomal copper concentration In WD patients was obviously lower than that of controls, and the addtion of vanadste or vincristine would change the copper transporting of microsomes obviously .Whenincubated with vincristine, levels of copper in microsomewere greatly increased, while incubated with vanadate, the copper corcentrations in microsome were obviously formed. The results indicating that there were WDproteins , the copper tmnsportion P-type ATPase in themicrosome of hepstocytes. WD patients possessed abnormal feed of WD protein in themicrosome, and the agonist might correct the defect of copper transportion by promoting the activity of coppertransportion P-type ATPase. CONCLUSION: Copper tmnsportion P-type ATPase plays animportant role in hepatocytic cop permetabolism.Dysfunction of hepatocytic WD protein copper transported might be one of the most important factors for WD.