Inhibitor of protein kinase N3 suppresses excessive bone resorption in ovariectomized mice
Inhibitor of protein kinase N3 suppresses excessive bone resorption in ovariectomized mice
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DOI：
10.1007/s00774-021-01296-1
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发表时间：
2022-01
影响因子：
3.3
通讯作者：
Shunsuke Uehara;H. Mukai;T. Yamashita;M. Koide;Kohei Murakami;N. Udagawa;Yasuhiro Kobayashi
中科院分区：
文献类型：
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作者：
Shunsuke Uehara;H. Mukai;T. Yamashita;M. Koide;Kohei Murakami;N. Udagawa;Yasuhiro Kobayashi
骨吸收的长期抑制抑制了新骨的形成，因为这些过程在生理性骨重建过程中是耦合的。迫切需要开发不抑制骨形成的抗骨吸收剂。我们以前证明，成熟破骨细胞中的Wnt 5a-Ror 2信号通过蛋白激酶N3（Pkn 3）促进骨吸收活性。据报道，p38 MAPK抑制剂SB 202190以低Ki值（0.004 μM）抑制Pkn 3。我们在此研究了SB 202190对破骨细胞分化和功能的影响，在体外和vivo.Materials和methodsBone骨髓细胞培养中存在的M-CSF和GST-Rankl分化成多核破骨细胞。破骨细胞用增加浓度的SB 202190处理。对于体内研究，对10周龄雌性小鼠进行卵巢切除术（OVX）。OVX小鼠腹腔内给予Pkn 3抑制剂在2毫克/公斤或车辆4周，骨量进行了分析micro-CT. ResultsSB 202190抑制破骨细胞培养的Pkn 3的自磷酸化。SB 202190通过抑制肌动蛋白环的形成，显著抑制破骨细胞培养物中吸收陷窝的形成。SB 202190降低了破骨细胞培养物中c-Src的活性，而不影响Pkn 3和c-Src之间的相互作用。SB 202190的治疗衰减OVX诱导的骨丢失，而不影响破骨细胞或骨形成的成骨细胞的数量。ConclusionsOur结果表明，Pkn 3有潜力作为治疗目标，由于骨吸收增加骨丢失。SB 202190有望成为开发新型抗骨吸收剂的先导化合物。
IntroductionThe long-term inhibition of bone resorption suppresses new bone formation because these processes are coupled during physiological bone remodeling. The development of anti-bone-resorbing agents that do not suppress bone formation is urgently needed. We previously demonstrated that Wnt5a-Ror2 signaling in mature osteoclasts promoted bone-resorbing activity through protein kinase N3 (Pkn3). The p38 MAPK inhibitor SB202190 reportedly inhibited Pkn3 with a low Ki value (0.004 μM). We herein examined the effects of SB202190 on osteoclast differentiation and function in vitro and in vivo.Materials and methodsBone marrow cells were cultured in the presence of M-csf and GST-Rankl to differentiate into multinucleated osteoclasts. Osteoclasts were treated with increasing concentrations of SB202190. For in vivo study, 10-week-old female mice were subjected to ovariectomy (OVX). OVX mice were intraperitoneally administered with a Pkn3 inhibitor at 2 mg/kg or vehicle for 4 weeks, and bone mass was analyzed by micro-CT.ResultsSB202190 suppressed the auto-phosphorylation of Pkn3 in osteoclast cultures. SB202190 significantly inhibited the formation of resorption pits in osteoclast cultures by suppressing actin ring formation. SB202190 reduced c-Src activity in osteoclast cultures without affecting the interaction between Pkn3 and c-Src. A treatment with SB202190 attenuated OVX-induced bone loss without affecting the number of osteoclasts or bone formation by osteoblasts.ConclusionsOur results showed that Pkn3 has potential as a therapeutic target for bone loss due to increased bone resorption. SB202190 is promising as a lead compound for the development of novel anti-bone-resorbing agents.