Department of Neurology, Affiliated Hospital of Integrated Traditional Chinese and Western Medicine.
Department of Neurology, Jiangsu Province Academy of Traditional Chinese Medicine.
J Nippon Med Sch. 2023;90(1):89-95. doi: 10.1272/jnms.JNMS.2023_90-114.
Oxidative stress is an important mechanism in liver ischemia/reperfusion (I/R) injury. Hepatocyte apoptosis and proliferation occur in parallel with liver I/R injury, and the degree of apoptosis and proliferation determines the effects on hepatocytes. Vitamin D receptor (VDR) can lessen liver I/R injury, but previous studies focused mostly on inflammation and immunity.
HO was used to induce hepatocyte injury. Before treatment with HO, Hep-3B cells were pretreated with paricalcitol (PC) and siRNA-VDR. Rapamycin and chloroquine were also applied in the study.
The number of apoptotic cells was measured with an annexin V (AV) -fluorescein isothiocyanate apoptosis detection kit. Expression of proteins was measured by western blotting. As compared with the HO+Hep-3B group, levels of AV/PI, cleaved caspase-3, and p62 were lower, and expression levels of Bcl-2, proliferating cell nuclear antigen, and VDR were higher, in the PC+HO+Hep-3B group. When the VDR gene was silenced by siRNA-VDR in the siRNA-VDR+HO+Hep-3B group, expressions of AV/PI, cleaved caspase-3, and p62 were upregulated, and expressions of Bcl-2, proliferating cell nuclear antigen, and VDR were downregulated, as compared with values for the siRNA-NC+HO+Hep-3B group. Treatment with rapamycin or chloroquine partially reversed the effect of PC and siRNA-VDR on apoptosis and proliferation.
VDR mediates hepatocyte apoptosis and proliferation through autophagy.
氧化应激是肝缺血/再灌注（I/R）损伤的一个重要机制。肝细胞凋亡和增殖与肝 I/R 损伤同时发生，凋亡和增殖的程度决定了对肝细胞的影响。维生素 D 受体（VDR）可以减轻肝 I/R 损伤，但以前的研究主要集中在炎症和免疫方面。
使用 HO 诱导肝细胞损伤。在用 HO 处理之前，用帕立骨化醇（PC）和 siRNA-VDR 预处理 Hep-3B 细胞。还应用雷帕霉素和氯喹进行了研究。
用 Annexin V（AV）-异硫氰酸荧光素凋亡检测试剂盒测量凋亡细胞的数量。通过 Western blot 测量蛋白质的表达。与 HO+Hep-3B 组相比，PC+HO+Hep-3B 组的 AV/PI、cleaved caspase-3 和 p62 水平较低，Bcl-2、增殖细胞核抗原和 VDR 的表达水平较高。在 siRNA-VDR+HO+Hep-3B 组中沉默 VDR 基因后，siRNA-VDR+HO+Hep-3B 组的 AV/PI、cleaved caspase-3 和 p62 的表达上调，Bcl-2、增殖细胞核抗原和 VDR 的表达下调，与 siRNA-NC+HO+Hep-3B 组相比。用雷帕霉素或氯喹处理可部分逆转 PC 和 siRNA-VDR 对凋亡和增殖的作用。
VDR 通过自噬介导肝细胞凋亡和增殖。
