The Journal of Practical Medicine >
To investigate the effect of aloperine on bone metabolism in osteoporotic mice based on autophagy and apoptosis mediated by Wnt/β⁃catenin signaling pathway
Received date: 2024-11-25
Online published: 2025-02-28
Objective To investigate the effects of aloperine （ALO） on bone metabolism in osteoporosis （OP） mice via Wnt/β?catenin signaling pathway?mediated autophagy and apoptosis. Methods Sixty mice were randomly allocated into six groups （n = 10 per group）： Sham group （sham?operated mice）， OP group （osteoporosis model induced by bilateral ovariectomy）， L?ALO group （OP mice intraperitoneally injected with 10 mg/kg aloperine）， M?ALO group （OP mice intraperitoneally injected with 20 mg/kg aloperine）， H?ALO group （OP mice intraperitoneally injected with 30 mg/kg aloperine）， and EV group （OP mice administered 0.09 mg/kg estradiol valerate）. Bone mineral density and microstructure of the tibia were assessed. Hematoxylin and eosin （HE） staining was performed to examine the morphology of tibial bone tissue. Serum levels of OCN， OPG， ALP， Ca， and P were measured using ELISA. Protein expression levels of LC3-Ⅱ， LC3-Ⅰ， Beclin?1， P62， Caspase?3， Caspase?9， Bax， Wnt3a， β?catenin， and C?Myc were analyzed by Western blot. Autophagosomes were visualized using immunofluorescence. Results Compared with the Sham group， the bone mineral density （BMD） and trabecular thickness in the OP group were significantly reduced， while trabecular separation， bone surface area， and volume were significantly increased （P < 0.05）. The levels of OPG， OCN， Ca， and P were significantly downregulated， whereas ALP levels were significantly upregulated （P < 0.05）. Additionally， the LC3?Ⅱ/LC3?Ⅰ ratio and expression levels of Beclin?1， Wnt3a， β?catenin， and C?Myc proteins were significantly decreased， while the expression levels of P62， Caspase?3， Caspase?9， and Bax proteins were significantly increased （P < 0.05）. Compared with the OP group， the L?ALO， M?ALO， and H?ALO groups exhibited significant increases in BMD and trabecular thickness， along with significant decreases in trabecular separation， bone surface area， and volume （P < 0.05）. The levels of OPG， OCN， Ca， and P were significantly upregulated， while ALP levels were significantly downregulated （P < 0.05）. Furthermore， the LC3?Ⅱ/LC3?Ⅰ ratio and expression levels of Beclin?1， Wnt3a， β?catenin， and C?Myc proteins were significantly increased， while the expression levels of P62， Caspase?3， Caspase?9， and Bax proteins were significantly decreased （P < 0.05）. In contrast， compared with the OP group， the EV group showed significant increases in BMD and trabecular thickness， as well as significant decreases in trabecular separation， bone surface area to volume ratio （P < 0.05）. The levels of OPG， OCN， Ca， and P were significantly upregulated， while ALP levels were significantly downregulated （P < 0.05）. Moreover， the LC3?Ⅱ/LC3?Ⅰ ratio and expression levels of Beclin?1， Wnt3a， β?catenin， and C?Myc proteins were significantly increased， while the expression levels of P62， Caspase?3， Caspase?9， and Bax proteins were significantly decreased （P < 0.05）. Conclusions Peanine may promote autophagy in osteoblasts and inhibit their apoptosis， thereby improving bone metabolism in OP mice. This effect may be mediated through the activation of the Wnt/β?catenin signaling pathway.
Key words： aloperine; osteoporosis; Wnt/β-catenin signaling pathway; autophagy; apoptosis
Zhizhou XIAO , Ying HUANG , Huawei. BIAN . To investigate the effect of aloperine on bone metabolism in osteoporotic mice based on autophagy and apoptosis mediated by Wnt/β⁃catenin signaling pathway[J]. The Journal of Practical Medicine, 2025 , 41(4) : 500 -508 . DOI: 10.3969/j.issn.1006-5725.2025.04.006
| 1 | GREGSON C L， ARMSTRONG D J， BOWDEN J， et al. UK clinical guideline for the prevention and treatment of osteoporosis ［J］. Arch Osteoporos， 2022， 17（1）： 58. doi:10.1007/s11657-022-01061-5 |
| 2 | ONIZUKA N， ONIZUKA T. Disparities in Osteoporosis Prevention and Care： Understanding Gender， Racial， and Ethnic Dynamics ［J］. Curr Rev Musculoskelet Med， 2024， 17（9）： 365-372. doi:10.1007/s12178-024-09909-8 |
| 3 | FISCHER V， HAFFNER-LUNTZER M. Interaction between bone and immune cells： Implications for postmenopausal osteoporosis ［J］. Semin Cell Dev Biol， 2022， 123： 14-21. doi:10.1016/j.semcdb.2021.05.014 |
| 4 | ZHANG Y W， CAO M M， LI Y J， et al. Fecal microbiota transplantation ameliorates bone loss in mice with ovariectomy-induced osteoporosis via modulating gut microbiota and metabolic function ［J］. J Orthop Translat， 2022， 37： 46-60. doi:10.1016/j.jot.2022.08.003 |
| 5 | CHANG Z， ZHANG P， ZHANG M， et al. Aloperine suppresses human pulmonary vascular smooth muscle cell proliferation via inhibiting inflammatory response ［J］. Chin J Physiol， 2019， 62（4）： 157-165. doi:10.4103/cjp.cjp_27_19 |
| 6 | HU R， CHEN L， CHEN X， et al. Aloperine improves osteoporosis in ovariectomized mice by inhibiting RANKL-induced NF-κB， ERK and JNK approaches ［J］. Int Immunopharmacol， 2021， 97： 107720. doi:10.1016/j.intimp.2021.107720 |
| 7 | WANG X， TIAN Y， LIANG X， et al. Bergamottin promotes osteoblast differentiation and bone formation via activating the Wnt/β-catenin signaling pathway ［J］. Food Funct， 2022， 13（5）： 2913-2924. doi:10.1039/d1fo02755g |
| 8 | CAI Y， SUN H， SONG X， et al. The Wnt/β-catenin signaling pathway inhibits osteoporosis by regulating the expression of TERT： An in vivo and in vitro study ［J］. Aging （Albany NY）， 2023， 15（20）： 11471-11488. |
| 9 | LI X， LU Y， WEN P， et al. Matrine restrains the development of colorectal cancer through regulating the AGRN/Wnt/β-catenin pathway ［J］. Environ Toxicol， 2023， 38（4）： 809-819. doi:10.1002/tox.23730 |
| 10 | 李永志，韩礼军，李智斌，等. 秦岭箭叶淫羊藿对骨质疏松大鼠骨代谢及胫骨骨微结构的影响［J］. 疑难病杂志，2024，23（8）：993-998，1001. doi:10.3969/j.issn.1671-6450.2024.08.019 |
| 11 | 罗兰兰，张宇静，任明诗，等. 杜仲汤对去卵巢大鼠骨质疏松症的影响及机制研究［J］. 中药新药与临床药理，2024，35（4）：461-468. doi:10.19378/j.issn.1003-9783.2024.04.002 |
| 12 | OH W T， YANG Y S， XIE J， et al. WNT-modulating gene silencers as a gene therapy for osteoporosis， bone fracture， and critical-sized bone defects ［J］. Mol Ther， 2023， 31（2）： 435-453. doi:10.1016/j.ymthe.2022.09.018 |
| 13 | WARREN J T， ZOU W， DECKER C E， et al. Correlating RANK ligand/RANK binding kinetics with osteoclast formation and function ［J］. J Cell Biochem， 2015， 116（11）： 2476-2483. doi:10.1002/jcb.25191 |
| 14 | ZADJALI F AL， BROOKS J， O'NEILL T W， et al. Experiences of postmenopausal osteoporosis： A narrative review ［J］. Disabil Rehabil， 2024， 46（5）： 828-840. doi:10.1080/09638288.2023.2169770 |
| 15 | TRéMOLLIERES F A， CHABBERT-BUFFET N， PLU-BUREAU G， et al. Management of postmenopausal women： Collège National des Gynécologues et Obstétriciens Fran?ais （CNGOF） and Groupe d'Etude sur la Ménopause et le Vieillissement （GEMVi） Clinical Practice Guidelines ［J］. Maturitas， 2022， 163： 62-81. doi:10.1016/j.maturitas.2022.05.008 |
| 16 | YUAN F， PENG W， YANG C， et al. Teriparatide versus bisphosphonates for treatment of postmenopausal osteoporosis： A meta-analysis ［J］. Int J Surg， 2019， 66： 1-11. doi:10.1016/j.ijsu.2019.03.004 |
| 17 | LIU Y， YU P， PENG X， et al. Hexapeptide-conjugated calcitonin for targeted therapy of osteoporosis ［J］. J Control Release， 2019， 304： 39-50. doi:10.1016/j.jconrel.2019.04.042 |
| 18 | REID I R， BILLINGTON E O. Drug therapy for osteoporosis in older adults ［J］. Lancet， 2022， 399（10329）： 1080-1092. doi:10.1016/s0140-6736(21)02646-5 |
| 19 | PASCHALIS E P， GAMSJAEGER S， HASSLER N， et al. Vitamin D and calcium supplementation for three years in postmenopausal osteoporosis significantly alters bone mineral and organic matrix quality ［J］. Bone， 2017， 95： 41-46. doi:10.1016/j.bone.2016.11.002 |
| 20 | CHEN L R， KO N Y， CHEN K H. Medical Treatment for Osteoporosis： From Molecular to Clinical Opinions ［J］. Int J Mol Sci， 2019， 20（9） ： 2213. doi:10.3390/ijms20092213 |
| 21 | CHOI D， CHOI S， CHANG J， et al. Exposure to oral bisphosphonates and risk of gastrointestinal cancer ［J］. Osteoporos Int， 2020， 31（4）： 775-782. doi:10.1007/s00198-020-05327-x |
| 22 | CHEN Y J， JIA L H， HAN T H， et al. Osteoporosis treatment： current drugs and future developments ［J］. Front Pharmacol， 2024， 15： 1456796. doi:10.3389/fphar.2024.1456796 |
| 23 | TAO X， YIN L， XU L， et al. Dioscin： A diverse acting natural compound with therapeutic potential in metabolic diseases， cancer， inflammation and infections ［J］. Pharmacol Res， 2018， 137： 259-269. doi:10.1016/j.phrs.2018.09.022 |
| 24 | CAO G， HU S， NING Y， et al. Traditional Chinese medicine in osteoporosis： From pathogenesis to potential activity ［J］. Front Pharmacol， 2024， 15： 1370900. doi:10.3389/fphar.2024.1370900 |
| 25 | MUHAMMAD T， SAKHAWAT A， KHAN A A， et al. Aloperine in combination with therapeutic adenoviral vector synergistically suppressed the growth of non-small cell lung cancer ［J］. J Cancer Res Clin Oncol， 2020， 146（4）： 861-874. doi:10.1007/s00432-020-03157-2 |
| 26 | YU H I， SHEN H C， CHEN S H， et al. Autophagy Modulation in Human Thyroid Cancer Cells following Aloperine Treatment ［J］. Int J Mol Sci， 2019， 20（21）： 5315. doi:10.3390/ijms20215315 |
| 27 | LIU J S， HUO C Y， CAO H H， et al. Aloperine induces apoptosis and G2/M cell cycle arrest in hepatocellular carcinoma cells through the PI3K/Akt signaling pathway ［J］. Phytomedicine， 2019， 61： 152843. doi:10.1016/j.phymed.2019.152843 |
| 28 | TAHIR M， ALI S， ZHANG W， et al. Aloperine： A Potent Modulator of Crucial Biological Mechanisms in Multiple Diseases ［J］. Biomedicines， 2022， 10（4）： 905. doi:10.3390/biomedicines10040905 |
| 29 | CHEN X， ZHI X， PAN P， et al. Matrine prevents bone loss in ovariectomized mice by inhibiting RANKL-induced osteoclastogenesis ［J］. FASEB J， 2017， 31（11）： 4855-4865. doi:10.1096/fj.201700316r |
| 30 | JIANG C， MA Q， WANG S， et al. Oxymatrine Attenuates Osteoclastogenesis via Modulation of ROS-Mediated SREBP2 Signaling and Counteracts Ovariectomy-Induced Osteoporosis ［J］. Front Cell Dev Biol， 2021， 9： 684007. doi:10.3389/fcell.2021.684007 |
| 31 | BRENT M B. Pharmaceutical treatment of bone loss： From animal models and drug development to future treatment strategies ［J］. Pharmacol Ther， 2023， 244： 108383. doi:10.1016/j.pharmthera.2023.108383 |
| 32 | YAMAMOTO H， ZHANG S， MIZUSHIMA N. Autophagy genes in biology and disease ［J］. Nat Rev Genet， 2023， 24（6）： 382-400. doi:10.1038/s41576-022-00562-w |
| 33 | DERETIC V. Autophagy in inflammation， infection， and immunometabolism ［J］. Immunity， 2021， 54（3）： 437-453. doi:10.1016/j.immuni.2021.01.018 |
| 34 | ZHANG L， GUO Y F， LIU Y Z， et al. Pathway-based genome-wide association analysis identified the importance of regulation-of-autophagy pathway for ultradistal radius BMD ［J］. J Bone Miner Res， 2010， 25（7）： 1572-1580. doi:10.1002/jbmr.36 |
| 35 | TANG N， ZHAO H， ZHANG H， et al. Effect of autophagy gene DRAM on proliferation， cell cycle， apoptosis， and autophagy of osteoblast in osteoporosis rats ［J］. J Cell Physiol， 2019， 234（4）： 5023-5032. doi:10.1002/jcp.27304 |
| 36 | ZHANG L， ZHENG Y L， WANG R， et al. Exercise for osteoporosis： A literature review of pathology and mechanism ［J］. Front Immunol， 2022， 13： 1005665. doi:10.3389/fimmu.2022.1005665 |
| 37 | LIU F， FANG F， YUAN H， et al. Suppression of autophagy by FIP200 deletion leads to osteopenia in mice through the inhibition of osteoblast terminal differentiation ［J］. J Bone Miner Res， 2013， 28（11）： 2414-2430. doi:10.1002/jbmr.1971 |
| 38 | TANG T， LIANG H， WEI W， et al. Aloperine targets lysosomes to inhibit late autophagy and induces cell death through apoptosis and paraptosis in glioblastoma ［J］. Mol Biomed， 2023， 4（1）： 42. doi:10.1186/s43556-023-00155-x |
| 39 | OBENG E. Apoptosis （programmed cell death） and its signals-A review ［J］. Braz J Biol， 2021， 81（4）： 1133-1143. doi:10.1590/1519-6984.228437 |
| 40 | BERTHELOOT D， LATZ E， FRANKLIN B S. Necroptosis， pyroptosis and apoptosis： An intricate game of cell death ［J］. Cell Mol Immunol， 2021， 18（5）： 1106-1121. doi:10.1038/s41423-020-00630-3 |
| 41 | RU J Y， WANG Y F. Osteocyte apoptosis： The roles and key molecular mechanisms in resorption-related bone diseases ［J］. Cell Death Dis， 2020， 11（10）： 846. doi:10.1038/s41419-020-03059-8 |
| 42 | CHANDRA A， RAJAWAT J. Skeletal Aging and Osteoporosis： Mechanisms and Therapeutics ［J］. Int J Mol Sci， 2021， 22（7）： 3553. doi:10.3390/ijms22073553 |
| 43 | WEINSTEIN R S， MANOLAGAS S C. Apoptosis and osteoporosis ［J］. Am J Med， 2000， 108（2）： 153-164. doi:10.1016/s0002-9343(99)00420-9 |
| 44 | XU Z， WANG P， WANG Z， et al. ER-β accelerates the process of primary osteoporosis by promoting VEGFA-mediated apoptosis of osteoblasts［J］. Genomics， 2023， 115（6）： 110743. doi:10.1016/j.ygeno.2023.110743 |
| 45 | MORIISHI T， FUKUYAMA R， MIYAZAKI T， et al. Overexpression of BCLXL in Osteoblasts Inhibits Osteoblast Apoptosis and Increases Bone Volume and Strength［J］. J Bone Miner Res， 2016， 31（7）： 1366-1380. doi:10.1002/jbmr.2808 |
| 46 | WONG S K， MOHAMAD N V， JAYUSMAN P A， et al. A Review on the Crosstalk between Insulin and Wnt/β-Catenin Signalling for Bone Health［J］. Int J Mol Sci， 2023， 24（15）： 12441. doi:10.3390/ijms241512441 |
| 47 | LIU J， XIAO Q， XIAO J， et al. Wnt/β-catenin signalling： Function， biological mechanisms， and therapeutic opportunities ［J］. Signal Transduct Target Ther， 2022， 7（1）： 3. doi:10.1038/s41392-021-00762-6 |
| 48 | VISWESWARAN M， POHL S， ARFUSO F， et al. Multi-lineage differentiation of mesenchymal stem cells-To Wnt， or not Wnt ［J］. Int J Biochem Cell Biol， 2015， 68： 139-147. doi:10.1016/j.biocel.2015.09.008 |
| 49 | CHENG B F， FENG X， GAO Y X， et al. Neural Cell Adhesion Molecule Regulates Osteoblastic Differentiation Through Wnt/β-Catenin and PI3K-Akt Signaling Pathways in MC3T3-E1 Cells ［J］. Front Endocrinol （Lausanne）， 2021， 12： 657953. doi:10.3389/fendo.2021.657953 |
| 50 | YU W， XIE C R， CHEN F C， et al. LGR5 enhances the osteoblastic differentiation of MC3T3-E1 cells through the Wnt/β-catenin pathway ［J］. Exp Ther Med， 2021， 22（2）： 889. doi:10.3892/etm.2021.10321 |
| 51 | ZHAO Y， LIU J， ZHANG Y， et al. Mir-381-3p aggravates ovariectomy-induced osteoporosis by inhibiting osteogenic differentiation through targeting KLF5/Wnt/β-catenin signaling pathway ［J］. J Orthop Surg Res， 2024， 19（1）： 480. doi:10.1186/s13018-024-04992-6 |
| 52 | LI R， RUAN Q， YIN F， et al. MiR-23b-3p promotes postmenopausal osteoporosis by targeting MRC2 and regulating the Wnt/β-catenin signaling pathway ［J］. J Pharmacol Sci， 2021， 145（1）： 69-78. doi:10.1016/j.jphs.2020.11.004 |
| 53 | XIAO X， AO M， XU F， et al. Effect of matrine against breast cancer by downregulating the vascular endothelial growth factor via the Wnt/β-catenin pathway ［J］. Oncol Lett， 2018， 15（2）： 1691-1697. |
/
| 〈 |
|
〉 |