Department of Orthopedics, Third People's Hospital of Shenzhen, No. 29 Bulan Road, Longgang, Shenzhen, 518112, Guangdong, People's Republic of China.
Department of Orthopedics, Shenzhen No. 2 People's Hospital, Shenzhen, 518000, People's Republic of China.
Mol Med. 2021 Feb 26;27(1):19. doi: 10.1186/s10020-021-00276-5.
Osteoporosis is a common phenomenon in HIV patients on tenofovir treatment, but its underlying mechanisms remain to be explored.
Quantitative real-time PCR was performed to analyze the expression of miR-302, miR-101, miR-145 and osteoclast-specific genes in the serum of HIV patients treated with tenofovir and ZOL. ELISA was used to evaluate the expression of RANKL, SMAD3 and PRKACB in the serum of these patients. Luciferase assay was carried out to explore the inhibitory effects of miR-302, miR-101 and miR-145 on the expression of PRKACB, RANKL and SMAD3, respectively. Western blot was used to examine the expression of genes involved in NF‑κB and JNK signaling pathways.
ZOL treatment significantly suppressed the expression of CTx and osteocalcin in HIV patients treated with tenofovir. The BMD loss of HIV patients treated with tenofovir was effectively hindered by ZOL treatment. Mechanistically, the expression of miR-302, miR-101, miR-145, RANKL, SMAD3 and PRKACB in the serum was remarkably activated by ZOL treatment. Luciferase assays showed that miR-302, miR-101 and miR-145 effectively suppressed the expression of PRKACB, RANKL and SMAD3, respectively, through binding to their 3' UTR. Furthermore, ZOL treatment notably restored the normal expression of osteoclast‑specific genes while activating NF‑κB and JNK signaling pathways.
The findings of this study demonstrated that administration of ZOL suppressed the expression of RANKL via modulating signaling pathways of miR-101-3p/RANKL, miR-302/PRKACB/RANKL and miR-145/SMAD3/RANKL. Furthermore, down-regulated expression of RANKL by ZOL treatment alleviated osteoporosis in HIV-positive subjects treated with tenofovir.
骨质疏松症是接受替诺福韦治疗的 HIV 患者的常见现象，但其中的潜在机制仍有待探索。
采用实时定量 PCR 分析接受替诺福韦和唑来膦酸治疗的 HIV 患者血清中 miR-302、miR-101、miR-145 和破骨细胞特异性基因的表达。采用 ELISA 法检测这些患者血清中 RANKL、SMAD3 和 PRKACB 的表达。通过荧光素酶报告基因实验分别探讨 miR-302、miR-101 和 miR-145 对 PRKACB、RANKL 和 SMAD3 表达的抑制作用。采用 Western blot 检测 NF-κB 和 JNK 信号通路相关基因的表达。
唑来膦酸治疗可显著抑制接受替诺福韦治疗的 HIV 患者 CTx 和骨钙素的表达。唑来膦酸治疗有效阻止了接受替诺福韦治疗的 HIV 患者的骨密度丢失。机制上，唑来膦酸治疗可显著激活接受替诺福韦治疗的 HIV 患者血清中 miR-302、miR-101、miR-145、RANKL、SMAD3 和 PRKACB 的表达。荧光素酶报告基因实验表明，miR-302、miR-101 和 miR-145 可通过结合其 3'UTR 有效抑制 PRKACB、RANKL 和 SMAD3 的表达。此外，唑来膦酸治疗还可显著恢复破骨细胞特异性基因的正常表达，同时激活 NF-κB 和 JNK 信号通路。
本研究结果表明，唑来膦酸通过调节 miR-101-3p/RANKL、miR-302/PRKACB/RANKL 和 miR-145/SMAD3/RANKL 信号通路抑制 RANKL 的表达，从而抑制其表达。此外，唑来膦酸治疗下调 RANKL 的表达可缓解接受替诺福韦治疗的 HIV 阳性患者的骨质疏松症。
