Jinkui Shenqi Pill accelerates osteoporotic fracture healing by promoting bone formation through neurosensory PGE2/EP4/p-CREB axis

PurposeJinkui Shenqi Pill (JKSQP), a traditional Chinese herbal formula, is clinically utilized in China for managing bone disorders secondary to kidney deficiency, including osteoporotic fractures (OPFs). The present study aims to elucidate the pharmacological mechanism underlying JKSQP’s therapeut...

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Main Authors: Jihao Xi, Danqing Fu, Dan Xu, Ruhan Shen, Yan Zhao, Haoqiang Dai, Chenjie Xia, Peihong Zhou
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-07-01
Series:Frontiers in Endocrinology
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Online Access:https://www.frontiersin.org/articles/10.3389/fendo.2025.1570685/full
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author Jihao Xi
Jihao Xi
Danqing Fu
Danqing Fu
Dan Xu
Ruhan Shen
Ruhan Shen
Yan Zhao
Haoqiang Dai
Haoqiang Dai
Chenjie Xia
Chenjie Xia
Peihong Zhou
author_facet Jihao Xi
Jihao Xi
Danqing Fu
Danqing Fu
Dan Xu
Ruhan Shen
Ruhan Shen
Yan Zhao
Haoqiang Dai
Haoqiang Dai
Chenjie Xia
Chenjie Xia
Peihong Zhou
author_sort Jihao Xi
collection DOAJ
description PurposeJinkui Shenqi Pill (JKSQP), a traditional Chinese herbal formula, is clinically utilized in China for managing bone disorders secondary to kidney deficiency, including osteoporotic fractures (OPFs). The present study aims to elucidate the pharmacological mechanism underlying JKSQP’s therapeutic effects on OPF healing.MethodsLC-MS/MS was employed to characterize the chemical constituents of JKSQP. Two-month-old female C57BL/6J mice underwent bilateral ovariectomy (OVX) followed by transverse tibial osteotomy to establish the OPF model. These OPF mice were randomly divided into the JKSQP group and OPF group, in which mice were gavaged with 1 g/kg/day JKSQP and equivalent volume of normal saline, respectively. At 4, 14, and 24 days post-fracture, biological specimens including serum, tibiae, dorsal root ganglion (DRG) and hypothalamus were collected for ELISA assay, μCT analysis and histopathology staining. Primary bone marrow stromal cells (BMSCs) were treated with the serum obtained from Sprague-Dawley rats administered with 1.5 g/kg/day JKSQP via oral gavage for three consecutive days. The conditioned medium derived from these JKSQP serum-treated BMSCs and the serum collected from the JKSQP-treated mice were applied to the DRG neurons. The levels of COX-2, PGE2, EP4 and CGRP in vitro were detected using qRT-PCR, western blot, ELISA and immunofluorescence (IF).ResultsLC-MS/MS analysis identified 1872 chemical components in JKSQP. μCT evaluation demonstrated accelerated healing of OPF in JKSQP-treated mice. Histomorphometric analysis combined with Calcein double-labeling revealed enhanced bone formation within the fracture callus. Compared with OPF controls, mice in the JKSQP group exhibited elevated serum PGE2 levels, upregulated Osterix, COX-2 and EP4 expression in fracture callus, increased EP4 and CGRP in DRG, and enhanced p-CREB in hypothalamus. In vitro, JKSQP-containing serum increased both PGE2 secretion and COX-2 expression in BMSCs. Furthermore, qRT-PCR and IF analyses confirmed that both conditioned medium from JKSQP-treated BMSCs and serum from JKSQP-administered mice upregulated EP4 and CGRP expressions in DRG neurons.ConclusionJinkui Shenqi Pill accelerates OPF healing by promoting bone formation possibly through activation of neurosensory PGE2/EP4/p-CREB axis.
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publisher Frontiers Media S.A.
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series Frontiers in Endocrinology
spelling doaj-art-2c41c12c2d13409da796e8f6d0191df92025-08-20T02:43:25ZengFrontiers Media S.A.Frontiers in Endocrinology1664-23922025-07-011610.3389/fendo.2025.15706851570685Jinkui Shenqi Pill accelerates osteoporotic fracture healing by promoting bone formation through neurosensory PGE2/EP4/p-CREB axisJihao Xi0Jihao Xi1Danqing Fu2Danqing Fu3Dan Xu4Ruhan Shen5Ruhan Shen6Yan Zhao7Haoqiang Dai8Haoqiang Dai9Chenjie Xia10Chenjie Xia11Peihong Zhou12Affiliated Cixi Hospital, Wenzhou Medical University, Ningbo, ChinaDepartment of Orthopedic Surgery, The Affiliated Lihuili Hospital of Ningbo University, Ningbo, ChinaAffiliated Cixi Hospital, Wenzhou Medical University, Ningbo, ChinaSchool of Basic Medical Sciences, Zhejiang Chinese Medical University, Hangzhou, ChinaSchool of Basic Medical Sciences, Zhejiang Chinese Medical University, Hangzhou, ChinaAffiliated Cixi Hospital, Wenzhou Medical University, Ningbo, ChinaSchool of Basic Medical Sciences, Zhejiang Chinese Medical University, Hangzhou, ChinaNingbo Municipal Hospital of Traditional Chinese Medicine, Affiliated Hospital to Zhejiang Chinese Medical University, Ningbo, ChinaAffiliated Cixi Hospital, Wenzhou Medical University, Ningbo, ChinaDepartment of Orthopedic Surgery, The Affiliated Lihuili Hospital of Ningbo University, Ningbo, ChinaAffiliated Cixi Hospital, Wenzhou Medical University, Ningbo, ChinaDepartment of Orthopedic Surgery, The Affiliated Lihuili Hospital of Ningbo University, Ningbo, ChinaNingbo Municipal Hospital of Traditional Chinese Medicine, Affiliated Hospital to Zhejiang Chinese Medical University, Ningbo, ChinaPurposeJinkui Shenqi Pill (JKSQP), a traditional Chinese herbal formula, is clinically utilized in China for managing bone disorders secondary to kidney deficiency, including osteoporotic fractures (OPFs). The present study aims to elucidate the pharmacological mechanism underlying JKSQP’s therapeutic effects on OPF healing.MethodsLC-MS/MS was employed to characterize the chemical constituents of JKSQP. Two-month-old female C57BL/6J mice underwent bilateral ovariectomy (OVX) followed by transverse tibial osteotomy to establish the OPF model. These OPF mice were randomly divided into the JKSQP group and OPF group, in which mice were gavaged with 1 g/kg/day JKSQP and equivalent volume of normal saline, respectively. At 4, 14, and 24 days post-fracture, biological specimens including serum, tibiae, dorsal root ganglion (DRG) and hypothalamus were collected for ELISA assay, μCT analysis and histopathology staining. Primary bone marrow stromal cells (BMSCs) were treated with the serum obtained from Sprague-Dawley rats administered with 1.5 g/kg/day JKSQP via oral gavage for three consecutive days. The conditioned medium derived from these JKSQP serum-treated BMSCs and the serum collected from the JKSQP-treated mice were applied to the DRG neurons. The levels of COX-2, PGE2, EP4 and CGRP in vitro were detected using qRT-PCR, western blot, ELISA and immunofluorescence (IF).ResultsLC-MS/MS analysis identified 1872 chemical components in JKSQP. μCT evaluation demonstrated accelerated healing of OPF in JKSQP-treated mice. Histomorphometric analysis combined with Calcein double-labeling revealed enhanced bone formation within the fracture callus. Compared with OPF controls, mice in the JKSQP group exhibited elevated serum PGE2 levels, upregulated Osterix, COX-2 and EP4 expression in fracture callus, increased EP4 and CGRP in DRG, and enhanced p-CREB in hypothalamus. In vitro, JKSQP-containing serum increased both PGE2 secretion and COX-2 expression in BMSCs. Furthermore, qRT-PCR and IF analyses confirmed that both conditioned medium from JKSQP-treated BMSCs and serum from JKSQP-administered mice upregulated EP4 and CGRP expressions in DRG neurons.ConclusionJinkui Shenqi Pill accelerates OPF healing by promoting bone formation possibly through activation of neurosensory PGE2/EP4/p-CREB axis.https://www.frontiersin.org/articles/10.3389/fendo.2025.1570685/fullJinkui Shenqi Pillsosteoporotic fracturebone formationsensory nervePGE2/EP4/p-CREB axis
spellingShingle Jihao Xi
Jihao Xi
Danqing Fu
Danqing Fu
Dan Xu
Ruhan Shen
Ruhan Shen
Yan Zhao
Haoqiang Dai
Haoqiang Dai
Chenjie Xia
Chenjie Xia
Peihong Zhou
Jinkui Shenqi Pill accelerates osteoporotic fracture healing by promoting bone formation through neurosensory PGE2/EP4/p-CREB axis
Frontiers in Endocrinology
Jinkui Shenqi Pills
osteoporotic fracture
bone formation
sensory nerve
PGE2/EP4/p-CREB axis
title Jinkui Shenqi Pill accelerates osteoporotic fracture healing by promoting bone formation through neurosensory PGE2/EP4/p-CREB axis
title_full Jinkui Shenqi Pill accelerates osteoporotic fracture healing by promoting bone formation through neurosensory PGE2/EP4/p-CREB axis
title_fullStr Jinkui Shenqi Pill accelerates osteoporotic fracture healing by promoting bone formation through neurosensory PGE2/EP4/p-CREB axis
title_full_unstemmed Jinkui Shenqi Pill accelerates osteoporotic fracture healing by promoting bone formation through neurosensory PGE2/EP4/p-CREB axis
title_short Jinkui Shenqi Pill accelerates osteoporotic fracture healing by promoting bone formation through neurosensory PGE2/EP4/p-CREB axis
title_sort jinkui shenqi pill accelerates osteoporotic fracture healing by promoting bone formation through neurosensory pge2 ep4 p creb axis
topic Jinkui Shenqi Pills
osteoporotic fracture
bone formation
sensory nerve
PGE2/EP4/p-CREB axis
url https://www.frontiersin.org/articles/10.3389/fendo.2025.1570685/full
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