CAJ | 학술논문

目的：观察乙肝病毒X蛋白（HBx）对肾小管上皮细胞凋亡的作用，并探讨其在乙型病毒肝炎相关性肾炎（HBVGN）发病中的分子机制。方法：将构建好的HBx真核表达载体pcDNA3．1（＋）-HBx转染至体外培养的人肾近曲小管上皮细胞（ HK-2细胞）中。 Western blotting 法检测转染后目的蛋白的表达及JAK2/STAT3信号通路的活化。细胞免疫荧光检测STAT3及p-STAT3表达水平。 CCK-8法检测细胞增殖活性。 Hoechst 33342染色观察细胞的形态学改变。透射电镜观察细胞的超微结构及凋亡。 Annexin V/PI双染流式细胞术检测细胞凋亡率。结果：转染目的基因HBx后，p-JAK2及p-STAT3表达水平均显著增加；同时，细胞增殖明显受抑。透射电镜、Ho-echst 33342染色及Annexin V/PI双染流式细胞术检测发现HBx促进HK-2细胞凋亡。 AG490（ JAK2/STAT3信号通路阻断剂）孵育后能够部分阻断JAK2/STAT3信号通路，减少HBx 所致细胞凋亡。结论：HBx 通过激活JAK2/STAT3信号通路导致肾小管上皮细胞凋亡，可能参与了HBV直接损伤肾组织的致病机制。
목적：관찰을간병독X단백（HBx）대신소관상피세포조망적작용，병탐토기재을형병독간염상관성신염（HBVGN）발병중적분자궤제。방법：장구건호적HBx진핵표체재체pcDNA3．1（＋）-HBx전염지체외배양적인신근곡소관상피세포（ HK-2세포）중。 Western blotting 법검측전염후목적단백적표체급JAK2/STAT3신호통로적활화。세포면역형광검측STAT3급p-STAT3표체수평。 CCK-8법검측세포증식활성。 Hoechst 33342염색관찰세포적형태학개변。투사전경관찰세포적초미결구급조망。 Annexin V/PI쌍염류식세포술검측세포조망솔。결과：전염목적기인HBx후，p-JAK2급p-STAT3표체수평균현저증가；동시，세포증식명현수억。투사전경、Ho-echst 33342염색급Annexin V/PI쌍염류식세포술검측발현HBx촉진HK-2세포조망。 AG490（ JAK2/STAT3신호통로조단제）부육후능구부분조단JAK2/STAT3신호통로，감소HBx 소치세포조망。결론：HBx 통과격활JAK2/STAT3신호통로도치신소관상피세포조망，가능삼여료HBV직접손상신조직적치병궤제。
AIM: To investigate the correlation of hepatitis B virus X protein (HBx) with renal tubular epithelialcell apoptosis in hepatitis B virus-associated glomerulonephritis (HBVGN) and the possible signaling mechanism. METHODS: The activation of JAK2/STAT3 signal pathway and the expression of apoptosis -related proteins in humankindey proximal tubular epithelial cells (HK-2 cells) were determined by Western blotting after transfection with HBx eukaryoticexpression vector.The cell proliferation was observed by CCK-8 assay.The cell apoptosis was analyzed by the imagingof HO33342 staining, transmission electron microscopy and flow cytometry with Annexin V /PI double staining.RESULTS:After transfection of the target gene HBx, the expression levels of both p-JAK2 and p-STAT3 were significantly increased.At the same time, the cell proliferation was obviously inhibited, and the apoptotic rate was increased.After incubationwith AG490, the JAK2/STAT3 signal pathway was partially blocked, and the cell apoptosis induced by HBx was reduced. CONCLUSION: HBx up-regulates the activation of JAK2/STAT3 signal pathway to induce renal tubular epithelialcell apoptosis, which is possibly involved in the pathogenic mechanism that HBV directly damages nephridial tissue .