Tanaka Kimiaki, Kobayashi Naoya, Gutierrez Alejandro Soto, Rivas-Carrillo Jorge David, Navarro-Alvarez Nalu, Chen Yong, Narushima Michiki, Miki Atsushi, Okitsu Teru, Noguchi Hirofumi, Tanaka Noriaki
Department of Surgery, Okayama University Graduate School of Medicine and Dentistry, Okayama, Japan.
Transplantation. 2006 Feb 15;81(3):427-37. doi: 10.1097/01.tp.0000188693.48882.18.
Because hepatocyte transplantation has been considered to be an attractive method to treat acute liver failure (ALF), efficient recovery of hepatocytes and maintenance of differentiated hepatocyte functions is of extreme importance. We here report the usefulness of an antiapoptotic pentapeptide V5, composed of Val-Pro-Met-Leu-Lys, in the monkey hepatocyte cultures.
We evaluated albumin production, metabolizing abilities of ammonia, lidocaine, and diazepam of monkey hepatocytes cultured with V5. The protein expression of apoptosis-associated molecules was analyzed using power blot analysis. An unwoven cloth inoculated with V5-treated monkey hepatocytes was transplanted on the surface of the spleen of both SCID mice and Balb/c mice suffering from ALF induced by 90% hepatectomy.
When 100 microM V5 was utilized, ammonia-, lidocaine- and diazepam- metabolizing capacities and albumin production ability were significantly increased in V5-treated monkey hepatocytes. Such hepatocytes showed decreased Annexin V binding and increased the expression of anti-apoptotic and/or cytoprotective molecules, including Ku70, NF-kappaB, IKAP, hILP/XIAP, IkappaB, and CAS. Transplantation of the cloth containing the monkey hepatocytes significantly improved blood levels of glucose and ammonia and encephalopathy score and prolonged the survival of the mice with ALF.
The present work clearly demonstrates the usefulness of V5 for maintaining the functions of monkey hepatocytes in tissue culture.
由于肝细胞移植被认为是治疗急性肝衰竭（ALF）的一种有吸引力的方法，因此高效恢复肝细胞并维持分化肝细胞的功能至关重要。我们在此报告一种由缬氨酸 - 脯氨酸 - 甲硫氨酸 - 亮氨酸 - 赖氨酸组成的抗凋亡五肽V5在猴肝细胞培养中的作用。
我们评估了用V5培养的猴肝细胞的白蛋白产生、氨、利多卡因和地西泮的代谢能力。使用免疫印迹分析凋亡相关分子的蛋白表达。将接种有V5处理的猴肝细胞的无纺布移植到经90%肝切除诱导急性肝衰竭的SCID小鼠和Balb/c小鼠的脾脏表面。
当使用100微摩尔V5时，V5处理的猴肝细胞中氨、利多卡因和地西泮的代谢能力以及白蛋白产生能力显著增加。这些肝细胞显示膜联蛋白V结合减少，抗凋亡和/或细胞保护分子的表达增加，包括Ku70、核因子κB、IKAP、人白细胞介素相关蛋白酶/ X连锁凋亡抑制蛋白、IκB和CAS。移植含有猴肝细胞的无纺布显著改善了血糖和氨的血液水平以及肝性脑病评分，并延长了急性肝衰竭小鼠的存活时间。
目前的工作清楚地证明了V5在组织培养中维持猴肝细胞功能的作用。
