1. Drynariae Rhizoma

Function of the herb:

• Strengthen myoskeletal systems

Scientific researches on Drynariae rhizome aka Rhizoma Gusuibu (the Chinese name translates to "Mender of Shattered Bones"), found that the herb assists and promotes the mending of sinews and bones. Drynariae Rhizoma (DR), a traditional Korea medicine, which is known for its effect to strengthen myoskeletal systems, frequently appears as the main ingredient in prescriptions for bone injuries. However, it is unclear how it pharmacologically contributes to the reformation of bone. In this study, the effect of DR on bone cells was investigated in vitro for the first time. The human osteoprecursor cells (OPC-1) were incubated in the medium with different concentrations of DR and the cell proliferation was studied. When the concentration of DR was < or = 120 microg ml(-1), the proliferation of OPC-1 was enhanced. However, the proliferation of OPC-1 was inhibited by DR with the concentrations of > 250 microg ml(-1). Under most treatments, the cells presented very pale expression for cyclooxygenase-2 protein; slightly intensified band showed at the highest DR concentration, 120 microg ml(-1) during the course of culture. On the other hand, we investigated the immunomodulatory activity of DR on cellular and humoral immunity. When different doses of ethanolic and water extracts of DR was administered to mice, it was dose-dependently potentiated the delayed-type hypersensitivity reaction induced by both sheep red blood cells (SRBC) and oxazolone. It significantly enhanced the production of circulating antibody titre in mice in response to SRBC. But, DR did not any effect on macrophage phagocytosis. Prolonged administration of DR significantly ameliorated the total white blood cell count and also restored the immunosuppressive effects induced by cyclophosphamide. The present investigation reveals that DR possesses immunomodulatory activity. From the results, it was concluded that DR directly stimulated the proliferation, alkaline phosphatase activity, protein secretion and particularly type I collagen synthesis of OPC-1 at dose-dependent manner, and stimulated both the cellular and the humoral immunity. “Effects of  Drynariae rhizoma on the proliferation of human bone cells and the immunomodulatory activity”. Department of Internal Medicine, Biochemistry and Molecular Biology, College of Oriental Medicine, Dongguk University and National Research Laboratory for Glycobiology, Sukjang-Dong 707, Kyungju, Kyungbuk 780-714, Republic of Korea. Pharmacol. Res. (2005). 29257

Effects of traditional Korean medicine, Drynariae rhizoma (DR), on the protease activity of bone loss-initiation in rats and mice were investigated. Ethanol extracts-DR (EE-DR) and water extracts-DR (WE-DR) were identified as potent inhibitor of cathepsins K and L. The original WE-DR inhibits cathepsins K and L with IC50 values of 3.7 microg/ml and 4.5 microg/ml, respectively. EE-DR was more potent than that of WE-DR, because the inhibitions of cathepsin K and L increased to 0.5 microg/ml and 0.8 microg/ml, respectively. The EE-DR was proved to be the most potent. EE-DR was found to be a potent inhibitor of cathepsins K with a Kin value of 5.0 microg/ml for cathepsin K. The activity was increased by 10-fold when the assay is performed in the presence of glutathioneat pH 7.0, which favors the formation of a GSH thiolate anion. Thus, it is suggested that this increase in potency is probably due to an enhanced chemical reactivity of the extract mixtures toward the thiolate of the active site of the enzyme. WE-DR exhibited time-dependet inhibition which allowed us to determine the association and dissociation rate constants with cathepsin K. Finally, EE-DR inhibitsbone resorption in an in vitro assay involving mouse osteocleasts and bovine bone with an IC50 value of 70 microg/ml. WE-DR represents a new herbal formulation inhibiting cathepsin K and L activity and proteolysis of bone collagen. These results strongly suggest that DR is effective for preventing the development of bone loss induced by cathepsin K. This result also suggested that the DR is effective for bone resorptive action in bone cells. “Inhibitory activity of Drynariae rhizoma extracts on cathespin having bone resorption activity”. Immunopharmacology and immunotoxicology. (2004) 18171

In the traditional Korean medicine, Drynariae Rhizoma (DR) [Drynaria fortunei (kunze) J. Sm] has been reported as a good enhancer for bone healing. In this experiment, we investigate the effects of DR on bone resorption using the bone cells culture. Different concentrations of crude extract of DR were added to mouse bone cells culture. The mitochondria activity of the bone cells after exposure was determined by colorimetric MTT assay. It was demonstrated that DR has potential effects on the bone cells culture without any cytotoxicity. The most effective concentration of DR on bone cells was 100 micro g/ml. On the other hand, cathepsin K (Cat K) is the major cysteine protease expressed in osteoclasts and is thought to play a key role in matrix degradation during bone resorption. In this study, Mouse long bone cells including osteoclasts and osteoblast were treated with the PI3-kinase inhibitor, wortmannin (WT), and a specific inhibitor of protein kinase C (PKC), calphostin C. Although WT prevented the osteoclast-mediated intracellular processing of Cat K, calphostin C did not. Similarly, treatment of osteoclasts-containing long bone cells with Drynariae Rhizoma (DR) extracts prevented the intracellular maturation of Cat K, suggesting that DR may disrupt the intracellular trafficking of pro Cat K. This is similar to that of WT. Since secreted proenzymes have the potential to reenter the cell via mannose-6-phosphate (M6P) receptor, to prevent this possibility, we tested WT and DR in the absence or presence of M6P. Inhibition of Cat K processing by WT or DR was observed in a dose-dependent manner. Furthermore, the addition of M6P resulted in enhanced potency of WT and DR. DR dose-dependently inhibited in vitro bone resorption with a potency similar to that observed for inhibition of Cat K processing. “Inhibition of Drynariae Rhizoma extracts on bone resorption mediated by processing of cathepsin K in cultured mouse osteoclasts”. Int Immunopharmacol 2003 Nov;3(12):1685-97 55293

Pharmacological factors are needed to prevent bone loss that occurs with increasing age. The chemical compounds that act on bone metabolism in herbal medicines, however, are poorly understood. Effects of traditional Korean medicine, Drynariae Rhizoma [Drynaria fortunei (kunze) J. Sm] extract (DR), on the osteoblastic proliferation and differentiation were investigated. The effect of DR, a natural phyto herb, on the proliferation and osteoblastic differentiation in non-transformed osteoblastic cells (MC3T3-E1) was studied. DR dose-dependently increased DNA synthesis (significant at 50–150 μg/ml). DR increased alkaline phosphatase (ALP) activity and prolyl hydroxylase activity of MC3T3-E1 cells (50–150 μg/ml). Antiestrogen tamoxifen eleminated the stimulation of proliferation and ALP activity of MC3T3-E1, which were induced by DR. DR at concentrations ranged from 30–100 μg/ml inhibited prostaglandin E2 production in MC3T3-E1. These results indicate that DR directly stimulates cell proliferation and differentiation of osteoblasts. These results also suggest and DR is effective for bone anti-resorptive action in bone cells. “Stimulative effects of Drynariae Rhizoma extracts on the proliferation and differentiation of osteoblastic MC3T3-E1 Cells” Elsevier Science Ltd Journal of Enthnopharmacology Volume 96, Issue 3, 15 January 2005, Pages 489-495 19569

In a previous study (Jeong et al., 2003, Inhibition of Drynariae Rhizoma extracts on bone resorption mediated by processing of cathepsin K in cultured mouse osteoclasts. International Immunopharmacology 3, 1685-1697), treatment of osteoclasts-containing long bone cells with Drynariae Rhizoma (DR) extract prevented the intracellular maturation of cathepsin K and thus, it was considered that DR is a pro-drug of a potent bone resorption inhibitor. To further clarify the role of DR in ossification, we investigated the effects of DR on the proliferation and differentiation of osteoblastic cell lines in vitro. In this study, the bone effect of DR is studied. We assessed the effects of DR on osteoblastic differentiation in nontransformed osteoblastic cells (MC3T3-E1) and rat bone marrow cells. DR enhanced alkaline phosphatase (ALP) activity and mineralization in a dose- and time-dependent fashion. This stimulatory effect of the DR was observed at relatively low doses (significant at 50-150 µg/ml and maximal at 150 µg/ml). Northern blot analysis showed that the DR (100 µg/ml) increased in bone morphogenetic protein-2 as well as ALP mRNA concentrations in MC3T3-E1 cells. DR (60 µg/ml) slightly increased in type I collagen mRNA abundance throughout the culture period, whereas it markedly inhibited the gene expression of collagenase-1 between days 15 and 20 of culture. These results indicate that DR has anabolic effects on bone through the promotion of osteoblastic differentiation, suggesting that it could be used for the treatment of common metabolic bone diseases such as osteoporosis. “Drynariae Rhizoma promotes osteoblast differentiation and mineralization in MC3T3-E1 cells through regulation of bone morphogenetic protein-2, alkaline phosphatase, type I collagen and collagenase-1”. Elsevier Science Ltd Toxicology in Vitro, 2004 (Vol. 18) (No. 6) 829-834 29834

2. Herba Cynomorium Songaricum

Function of the herb:

•  To reinforce the vital function of kidney
•  For premature ejaculation

In order to exploit the medicinal resources and provide a scientific evidence for the clinical use and quality of Cynomorium songaricum control, the chemical constituents and pharmacological effects of the stem of the plant were studied preliminarily. METHOD: The chemical constituents were studied by spectral analysis and chemical method. The pharmacological effects were studied by an experiment on the defecation and deficiency of kidney function. RESULT AND CONCLUSION: Seven compounds were isolated from the stem of C. songaricum and identified as beta-sitosterol(I), palmitic acid (II), ursloic acid (III), daucosterol (IV), (-)-catechin (VI), naringenin-4'-O-pyranogluoside (VII) and succinicyacid (XII). Compounds VI and XII were isolated from this species for the first time. The pharmacological results show that the different extracts facilitate catharsis and replenish the deficiency of kidney yang. “Studies on chemical constituents and pharmacological effects of the stem of Cynomorium songaricum Rupr”. Zhongguo Zhong Yao Za Zhi 1999 May;24(5):292-4, 318-9. 05886

3.  Cortex Eucommia Ulmoides

Function of the herb:

• Used to replenish liver and kidney
• Strengthen bones and muscles
• Treats aching back and knees
• Tone up liver and kidneys
• Strengthen tendon and bones

The most interesting study was conducted on an herb called Eucommia. Eucommia contains chemicals that fit into the testosterone receptor and cause it to fire as if testosterone itself were present - causing androgenic, or male-creating changes. These phytoandrogens also increase the activity of real testosterone. The researchers describe a phenomenal synergy between chemicals in Eucommia, testosterone receptors, and testosterone. (see abstract of the study below). Eucommia's value for strengthening the male system is not news to Asian herbalists. The Chinese have been using it for millenia, and they call it Du Zhong. They use it for increasing libido and treating impotence, as well as for some kidney/adrenal complaints, high blood pressure, arthritis and connective tissue diseases. It is one of the 50 main herbs of Traditional Chinese Medicine (TCM), and it has a long history of safe use. The history of searching for plant testosterone is an interesting look into what scientists know, and what they think they know. The scientific literature of the 1950s is littered with failed attempts to find plant sources of testosterone. They all failed because they were looking for a molecule that was identical to human testosterone. No mamalian testosterone was found in plants, but plants do produce some other molecules that hit the testosterone receptors. It took a laboratory 'bioassay' to convince scientists of what the TCM practitioners had been saying all along. The study on Eucommia found that it contains two classes of compounds that pump up Testosterone activity - terpenoids and an unusual type of fatty acids. Therefore it is useful for strengthening weak limps.

BACKGROUND: Plants containing compounds such as the isoflavonoids, with female hormone-like effects that bind to human estrogen receptors, are known. But none has been previously shown to have corresponding male hormone-like effects that interact with the human androgen receptor. Here, we report that the tree bark (cortex) of the Gutta-Percha tree Eucommia ulmoides possesses bimodal phytoandrogenic and hormone potentiating effects by lipidic components. METHODS: The extracts of E. ulmoides were tested using in-vitro reporter gene bioassays and in-vivo animal studies. Key compounds responsible for the steroidogenic effects were isolated and identified using solid phase extraction (SPE), high performance liquid chromatography (HPLC), thin layer chromatography (TLC), gas chromatography-mass spectroscopy (GC-MS), electron spray ionisation-mass spectroscopy (ESI-MS) and nuclear magnetic resonance (NMR). RESULTS: The following bioactivities of E. ulmoides were found: (1) a phenomenal tripartite synergism exists between the sex steroid receptors (androgen and estrogen receptors), their cognate steroidal ligands and lipidic augmenters isolated from E. ulmoides, (2) phytoandrogenic activity of E. ulmoides was mediated by plant triterpenoids binding cognately to the androgen receptor (AR) ligand binding domain. CONCLUSION: In addition to well-known phytoestrogens, the existence of phytoandrogens is reported in this study. Furthermore, a form of tripartite synergism between sex steroid receptors, sex hormones and plant-derived lipids is described for the first time. This could have contrasting clinical applications for hypogonadal- and hyperlipidaemic-related disorders. “Novel phytoandrogens and lipidic augmenters from Eucommia ulmoides”. BMC Complimentary Altern Med 2007 Jan 29;7:3 PMID: 17261169

The biologically active compounds and free radical-/ or reactive oxygen species (ROS)-/ scavenging effect of water extract from Du-zhong (WEDZ) were investigated. The WEDZ used included leaves, raw cortex, and roasted cortex. The hot water extract of Du-zhong leaves showed marked activity as a ROS scavenger, and the scavenging effect was concentration dependent. The extract of roasted cortex exhibited a modest scavenging effect on ROS, while the extract of raw cortex had the weakest scavenging effect. The scavenging activity of WEDZ on ROS was correlated to its protocatechuic acid (PCA) content. The content of PCA in Du-zhong determined by HPLC followed the order of leaves (17.17 mg/g) > roasted cortex (2.99 mg/g) > raw cortex (1.16 mg/g). The inhibitory activity of leaf extract of Du-zhong was stronger than that of PCA on the peroxidation of linoleic acid at the same concentration of 0.1 mg/mL. The results presented herein indicated that extract of Du-zhong could possibly. act as a prophylactic (remedy that prevents or slows the course of an illness or disease) agent to prevent free radical-related diseases. “Reactive oxygen species scavenging activity of Du-zhong (Eucommia ulmoides oliv.) and its active compounds”. J Agric Food Chem. 2000 Aug;48(8):3431-6. PMID: 10956129

4. Radix Gentiana Macrophylla

Function of the herb:

• Antirheumatic agent for treatment of rheumatic pain of the whole body
• For allergic inflammation
• Tones up the liver
• Strengthen bones, tendons and ligaments

Gentiana macrophylla is categorized in the Materia Medica with herbs that expel wind-dampness. The liver is said to nourish the tendons and ligaments and Gentiana macrophylla is particularly used in cases where these connective tissues become hypercontracted due to insufficient nourishment (e.g., deficiency of liver yin), yielding spasms that run from the joints to the muscles. Vangermeersch L and Sun Peilin, Bi-Syndromes, 1994 SATAS, Brussels, Belgium.

Gentiana macrophylla is as often indicated for "bone fever" in TCM pharmacology. This strange term refers to syndromes with fever and sweating in which the person feels as if there is steam coming from the bones, heating up the skin. It is used by herbal scientists to nourish and strenghten bones, temdons and ligaments. State Administration of Traditional Chinese Medicine, Advanced Textbook on Traditional Chinese Medicine and Pharmacology, (vol. 1) 1995-6 New World Press, Beijing.