Cistanche Deserticola Extract Increases Bone Formation in Osteoblasts--Part II
Mar 15, 2022
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Te-Mao Lia et al
Discussion
Cistanche deserticola Ma, a native herb in China, is widely used in traditional medicine for various therapeutic treatments due to its sedative, analgesic, and immunostimulatory activity.[11–15] Here, we showed that CD (Cistanche deserticola) extract induced bone mineralization in cultured osteoblasts, but did not affect their cell migration or proliferation. In addition, we found that ALP, BMP-2, and OPN are target proteins of CD (Cistanche deserticola) extract-induced signaling, which requires activation of ERK, p38, JNK, and NF-kB. Bone is a complex tissue composed of several cell types which are continuously undergoing a process of renewal and repair.[30] When resorption and formation of bone are imbalanced, bone breakdown overrides bone formation, and osteoporosis results.[30] We used ovariectomized mice to examine the anti-osteoporotic effect of CD (Cistanche deserticola) extract. Ovariectomized mice had reduced total body bone mineral density and bone mineral content, which was alleviated by treatment
with CD (Cistanche deserticola) extract. CD (Cistanche deserticola) extract also increased serum levels of the osteogenic marker ALP and osteocalcin. Therefore, the CD (Cistanche deserticola) is a bone formation agent that prevents bone loss brought about by ovariectomy in vivo.
Although the mechanisms of osteoporosis are not entirely clear, they are likely related to the decreased availability, or decreased effects, of bone growth factors, such as ALP, BMP-2, and OPN. These three factors play important roles in the process of bone formation and remodeling,[31] and it has been well documented that stimulation of osteoblast cell differentiation is characterized mainly by increased expression of ALP, BMP-2, and OPN.[32] In this study, we found that CD (Cistanche deserticola) extract increased ALP, BMP-2, and OPN expression and enhanced bone mineralization. Therefore, CD (Cistanche deserticola) extract mediates bone formation in part by upregulating the expression of ALP, BMP-2, and OPN.
It has been reported that p38 is involved in the regulation of ALP expression during the differentiation of osteoblastic cells;[33] similarly, ERK1/2 is important for the proliferation and differentiation of osteoblasts.[34,35] JNK is involved in osteoclast formation.[36] We showed here that CD (Cistanche deserticola) extract induced ERK, p38, and JNK phosphorylation, and inhibitors of these enzymes antagonized the CD (Cistanche deserticola) extract-mediated potentiation of bone mineralization, suggesting that ERK, p38, and JNK activation play an obligatory role in CD (Cistanche deserticola) extract induced bone formation by osteoblasts. In addition, the enzyme inhibitors and dominant-negative mutants of ERK, p38, and JNK reduced CD (Cistanche deserticola) extract-enhanced ALP, BMP-2, and OPN expression. These data suggest that activation of the ERK, p38, and JNK pathways is required for the increase of ALP, BMP-2, and OPN expression and maturation caused by CD extract in osteoblasts. ERK has been reported to increase osteoblast proliferation and differentiation.[34,35] However, we did not detect an effect of CD (Cistanche deserticola) extract on osteoblast proliferation. Therefore, other pathways may have counteracted the ERK effect after CD (Cistanche deserticola) extract stimulation, or be necessary for proliferation. In this regard, we have found that PI3K and Akt inhibitors also reduce CD extract-induced bone mineralization and ALP, BMP-2, or OPN mRNA expression (data not shown). Therefore, these pathways may be required for CD extract-induced bone formation.
Cistanche deserticola is good for bone formation
NF-kB has been shown to control osteoblast function in bone.[37] The results of this study show that NF-kB activation contributes to CD (Cistanche deserticola) extract-induced bone mineralization and ALP, BMP-2, and OPN expression in cultured osteoblasts and that inhibitor of the NF-kB-dependent signaling pathway (PDTC and TPCK) inhibited CD (Cistanche deserticola) extract-induced bone mineralization and ALP, BMP-2, and OPN expression. p65 is phosphorylated at Ser536 by a variety of kinases in several signaling pathways, which enhances the p65 transactivation potential.[38] The results of this study showed that CD (Cistanche deserticola) extract increased the phosphorylation of p65. Taken together, these results suggest that NF-kB activation is required for CD extract-induced bone formation in cultured osteoblasts. We also found that ERK, p38, and JNK inhibitors reversed CD (Cistanche deserticola) extract-induced NF-kB luciferase activity (data not shown), consistent with these enzymes being upstream mediators of CD extract-induced NF-kB activation. Therefore, CD (Cistanche deserticola) extract induces bone formation through the ERK/p38/JNK/ and NF-kB pathways.
Conclusions
This study demonstrated that CD (Cistanche deserticola) extract induces osteoblast differentiation and maturation but not proliferation or migration. CD (Cistanche deserticola) extract also increased ALP, BMP-2, and OPN expression and bone mineralization. We showed that the ERK, p38, JNK, and NF-kB pathways are involved in CD (Cistanche deserticola) extract-mediated bone formation and ALP, BMP-2, and OPN expression. Furthermore, CD (Cistanche deserticola) extract prevented in-vivo bone loss induced by ovariectomy. Therefore, CD (Cistanche deserticola) may be beneficial in stimulating bone formation in the treatment of osteoporotic diseases.
Cistanche deserticola is good for bone formation
Declarations
Conflict of interest
The Author(s) declare(s) that they have no conflicts of interest to disclose.
Funding
This work was supported by grants from the National Science Council of Taiwan (NSC100-2320-B-039-032; NSC100-2320-B-039-028-MY3).
Acknowledgments
We thank Dr. J. Han for providing the p38 mutant, Dr. M. Karin for providing the JNK mutant, and Dr. M. Cobb for providing the ERK2 mutant.
From: 'Cistanche deserticola extract increases bone formation in osteoblasts' by Te-Mao Lia et al
---© 2012 The Authors. JPP © 2012 Royal Pharmaceutical Society 2012 Journal of Pharmacy and Pharmacology, 64, pp. 897–907
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