Assessment of cytochrome P450 induction in canine intestinal organoid models

Understanding cytochrome P450 (CYP) enzymes in the canine intestine is vital for predicting drug metabolism and developing safer oral medications. This study evaluates canine colonoids as a model to assess the expression and induction of essential intestinal CYP enzymes.Canine colonoids were cultured in expansion medium (EM) with Wnt-3A and in differentiation medium (DM) without Wnt-3A. We assessed the mRNA expression of <i>CYP2B11</i>, <i>CYP2C21</i>, <i>CYP3A12</i>, and <i>CYP3A98</i> using qPCR and examined the effects of rifampicin and phenobarbital as inducers.Our findings show that DM significantly increased the mRNA expression of <i>CYP3A98</i> and <i>CYP2B11</i>, but not <i>CYP3A12</i>, compared to EM. <i>CYP2C21</i>, not typically expressed in the intestine, remained unexpressed in colonoids. Rifampicin induced CYP3A98, aligning with pregnane x receptor (PXR) regulation, while phenobarbital did not, suggesting no constitutive androstane receptor (CAR) involvement. CYP2B11 did not respond to either inducer, suggesting alternative regulatory pathways in canine colonoids.This study is a pioneering effort to establish conditions for studying P450 expression in canine colonoids, confirming significant CYP3A98 expression in the canine intestine. It demonstrated colonoids can induce CYP activity post drug treatments. Further research is needed to enhance species-specific drug metabolism understanding and validate this model for broader applications. Understanding cytochrome P450 (CYP) enzymes in the canine intestine is vital for predicting drug metabolism and developing safer oral medications. This study evaluates canine colonoids as a model to assess the expression and induction of essential intestinal CYP enzymes. Canine colonoids were cultured in expansion medium (EM) with Wnt-3A and in differentiation medium (DM) without Wnt-3A. We assessed the mRNA expression of <i>CYP2B11</i>, <i>CYP2C21</i>, <i>CYP3A12</i>, and <i>CYP3A98</i> using qPCR and examined the effects of rifampicin and phenobarbital as inducers. Our findings show that DM significantly increased the mRNA expression of <i>CYP3A98</i> and <i>CYP2B11</i>, but not <i>CYP3A12</i>, compared to EM. <i>CYP2C21</i>, not typically expressed in the intestine, remained unexpressed in colonoids. Rifampicin induced CYP3A98, aligning with pregnane x receptor (PXR) regulation, while phenobarbital did not, suggesting no constitutive androstane receptor (CAR) involvement. CYP2B11 did not respond to either inducer, suggesting alternative regulatory pathways in canine colonoids. This study is a pioneering effort to establish conditions for studying P450 expression in canine colonoids, confirming significant CYP3A98 expression in the canine intestine. It demonstrated colonoids can induce CYP activity post drug treatments. Further research is needed to enhance species-specific drug metabolism understanding and validate this model for broader applications.

阐明犬肠道内细胞色素P450（cytochrome P450, CYP）酶的功能，对预测药物代谢过程、研发更安全的口服药物至关重要。本研究以犬结肠类器官为模型，评估肠道核心CYP酶的表达与诱导调控情况。研究中将犬结肠类器官分别置于添加Wnt-3A的扩增培养基（expansion medium, EM）以及不含Wnt-3A的分化培养基（differentiation medium, DM）中进行培养。本研究通过实时荧光定量聚合酶链式反应（qPCR）检测了<i>CYP2B11</i>、<i>CYP2C21</i>、<i>CYP3A12</i>及<i>CYP3A98</i>的mRNA表达水平，并考察了利福平与苯巴比妥作为诱导剂的调控效果。研究结果显示，与EM组相比，DM组可显著提升<i>CYP3A98</i>与<i>CYP2B11</i>的mRNA表达水平，但对<i>CYP3A12</i>无此效果。<i>CYP2C21</i>通常不在肠道中表达，因此在结肠类器官中也未检测到其表达。利福平可诱导<i>CYP3A98</i>的表达，这与孕烷X受体（pregnane X receptor, PXR）的调控机制相符；而苯巴比妥无此诱导效果，提示组成型雄烷受体（constitutive androstane receptor, CAR）未参与该调控过程。<i>CYP2B11</i>对两种诱导剂均无响应，提示犬结肠类器官中存在其他调控通路。本研究为建立犬结肠类器官的CYP酶表达研究体系做出了开创性尝试，证实了犬肠道中存在显著的<i>CYP3A98</i>表达。本研究证实，结肠类器官可在药物处理后诱导CYP酶活性。未来仍需开展进一步研究，以加深对物种特异性药物代谢的理解，并验证该模型的更广泛应用潜力。
阐明犬肠道内细胞色素P450（cytochrome P450, CYP）酶的功能，对预测药物代谢过程、研发更安全的口服药物至关重要。本研究以犬结肠类器官为模型，评估肠道核心CYP酶的表达与诱导调控情况。研究中将犬结肠类器官分别置于添加Wnt-3A的扩增培养基（expansion medium, EM）以及不含Wnt-3A的分化培养基（differentiation medium, DM）中进行培养。本研究通过实时荧光定量聚合酶链式反应（qPCR）检测了<i>CYP2B11</i>、<i>CYP2C21</i>、<i>CYP3A12</i>及<i>CYP3A98</i>的mRNA表达水平，并考察了利福平与苯巴比妥作为诱导剂的调控效果。研究结果显示，与EM组相比，DM组可显著提升<i>CYP3A98</i>与<i>CYP2B11</i>的mRNA表达水平，但对<i>CYP3A12</i>无此效果。<i>CYP2C21</i>通常不在肠道中表达，因此在结肠类器官中也未检测到其表达。利福平可诱导<i>CYP3A98</i>的表达，这与孕烷X受体（pregnane X receptor, PXR）的调控机制相符；而苯巴比妥无此诱导效果，提示组成型雄烷受体（constitutive androstane receptor, CAR）未参与该调控过程。<i>CYP2B11</i>对两种诱导剂均无响应，提示犬结肠类器官中存在其他调控通路。本研究为建立犬结肠类器官的CYP酶表达研究体系做出了开创性尝试，证实了犬肠道中存在显著的<i>CYP3A98</i>表达。本研究证实，结肠类器官可在药物处理后诱导CYP酶活性。未来仍需开展进一步研究，以加深对物种特异性药物代谢的理解，并验证该模型的更广泛应用潜力。