Comparative study of CYP2B1/2 induction and the transport of bilirubin and taurocholate in rat hepatocyte-mono- and hepatocyte-Kupffer cell co-cultures

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• 作者：Attila Bá ; tai-Konczos^a ; ^{flax84@gmail.com" class="auth_mail" title="E-mail the corresponding author} ; Zsuzsa Veres^a ; ^{veres.zsuzsanna@ttk.mta.hu" class="auth_mail" title="E-mail the corresponding author} ; Mó ; nika Szabó ; ^a ; ^{szabo.monika@ttk.mta.hu" class="auth_mail" title="E-mail the corresponding author} ; Enikő Ioja^a ; ^{ioja.eniko@ttk.mta.hu" class="auth_mail" title="E-mail the corresponding author} ; Gló ; ria Lá ; szló ; ^b ; ^{glorial@caesar.elte.hu" class="auth_mail" title="E-mail the corresponding author} ; Gyö ; rgy Tö ; rö ; k^c ; ^{torok.gyorgy@ttk.mta.hu" class="auth_mail" title="E-mail the corresponding author} ; Lá ; szló ; Homolya^c ; ^{homolya.laszlo@ttk.mta.hu" class="auth_mail" title="E-mail the corresponding author} ; Katalin Jemnitz^a ; ^{jemnitz.katalin@ttk.mta.hu" class="auth_mail" title="E-mail the corresponding author}
• 关键词：B ; bilirubin ; Bsep ; bile salt export pump ; CYP ; cytochrome P-450 ; GdCl3 ; gadolinium(III) chloride ; H/KC ; hepatocyte-Kupffer-cell co-culture ; HBSS ; Hanks' balanced salt solution ; HPLC ; high-performance liquid chromatography ; KC ; Kupffer-cell ; LPS ; lipopolisacharide ; Mrp ; multidrug resistance-associated protein ; Ntcp ; sodium-dependent taurocholate transporter ; PB ; phenobarbital ; PBS ; phosphate-buffered saline ; PTX ; pentoxyphylline ; TC ; taurocholate
• 刊名：Journal of Pharmacological and Toxicological Methods
• 出版年：2016
• 出版时间：November-December 2016
• 年：2016
• 卷：82
• 期：Complete
• 页码：1-8
• 全文大小：903 K

文摘

Hepatocyte-Kupffer cell (KC) co-cultures represent a promising approach for in vitro modeling of complex interactions between parenchymal and non-parenchymal cells in the liver, responsible for drug-induced liver injury (DILI). In this study we aimed to compare hepatocyte monocultures with hepatocyte-KC co-cultures regarding some basic liver functions associated with the chemical defense system. These pathways involve transporters and enzymes the function of which is highly sensitive towards hepatotoxic events.

Methods

CYP2B1/2 induction and the biliary and sinusoidal elimination of bilirubin (B) and taurocholate (TC) were studied in rat hepatocyte sandwich cultures compared with rat hepatocyte-KC sandwich co-cultures of 1:0, 6:1, 2:1 and 1:1 cell combinations representing the physiologic and pathologic conditions of the liver.

Results

KCs decreased phenobarbital inducibility of CYP2B1/2 in a cell ratio dependent manner and activation of KCs by lipopolisacharide (LPS) amplified this effect. Similarly, KCs decreased the transport of B and its glucuronides (BG) in both sinusoidal and canalicular directions resulting in its intracellular accumulation. In contrast, the uptake and the efflux of TC were greater in the co-cultures than in the hepatocyte monocultures. Immuno-labelling of sodium-dependent taurocholate transporter (Ntcp) revealed increased expression of the transporter in the presence of KCs.

Discussion

Here we presented that KCs have a direct impact on some hepatocyte functions suggesting that the co-culture model may be more suitable for drug related hepatotoxicity studies than hepatocyte monocultures.