PART 2 Cistanche Tubulosa (Schenk) Wight Extract Enhances Hindlimb Performance And Attenuates Myosin Heavy Chain IId/IIx Expression in Cast-Immobilized Mice

CLICK HERE TO PART 1

For more information please contact: Joanna.jia@wecistanche.com

4. Discussion

Our novel cast immobilization method resulted in prominent muscle wasting, comparable to other cast immobilization methods [17–24], inducing approximately 10–45% of gastrocnemius muscle mass loss in 10 to 21 days. Also, myofiber CSA was decreased, indicating a reduction in muscle mass. This muscle change was accompanied by an increase of MyHC IId/IIx expression, which has been observed in several other studies of disuse muscle atrophy [11, 12]; hence, we believe that hindlimb disuse due to cast immobilization induced hindlimb muscle wasting.

The mammalian skeletal muscle contains slow-twitch and fast-twitch muscle fibers. The properties of these fibers are determined by the expression of multiple isoforms of MyHC (Iβ, IIa, IId/IIx, and IIb, which exist only in small mammals). Isoform Iβ has a slow-twitch profile, while isoforms IIa, IId/IIx, and IIb, in this order, have increasingly fast-twitch profiles, IIb being the quickest [36]. When muscle wasting occurs, a shift in the myofiber type also takes place, and, specifically, a slow-to-fast shift occurs in cases of loss of neural influence and mechanical loading. The changes in MyHC IId/IIx expression mentioned above are assumed to be a consequence of the myofiber type shift from slow to fast [10]. In contrast, malnutrition, inflammation, and sarcopenia induce a fast-to-slow myofiber type shift [10] through the reduction of type II fibers [1]; however, several reports revealed that type I fibers are also reduced in sarcopenia, where they form MyHC-coexpressing fibers [37]. Oral administration of CTE for 13 days tended to im- prove the deterioration of hindlimb function and to suppress the increase of MyHC IId/IIx expression, although not in a statistically significant way. Interestingly, muscle mass re- duction did not simultaneously improve. These results suggest that CTE can attenuate the slow-to-fast myofiber type shift, thus contrasting the deterioration of hindlimb muscle function.

Cistanche can improve hindlimb function, click here to know more

Since the TS presents naturally a slow-type contraction profile, it was suggested that slow-to-fast muscle fiber type shift along with muscle atrophy can induce impairment of muscle function, and an attenuation of this shift can mitigate it. For instance, Desaphy et al. reported that in the soleus muscle of hindlimb-unloaded mice, MyHC IId/IIx and IIb expression level increased, whereas that of IIa was decreased, and these changes occurred together with a shift of the mechanical threshold and alterations of the excitability parameters, along with muscle mass reduction; it was also revealed that antioxidant treatment by Trolox could

neutralize these changes, but there was no effect on muscle mass [11]. Ferraro et al. also revealed a similar phenomenon using trimetazidine (TMZ); TMZ administration in aged mice improved forelimb muscle strength without increasing muscle mass, and this change was accompanied by an increase in MyHC Iβ [33]. In contrast, 8-week administration of β2-adrenoreceptor BRL-47672 significantly increased MyHC IIb, significantly decreased MyHC Iβ expression, and led to a 22% increase of gastrocnemius-plantaris-soleus muscle complex (GPS) mass, although it resulted in the deterioration of GPS performance [38].

Cistanche is a precious Chinese herbal medicine, which has many effects on the human body

The molecular mechanisms underlying CTE’s effects on muscle wasting remain unknown, including the molecular components of CTE that affect muscle performance. We recently reported that at least one of the constituents of CTE, acteoside, ameliorates muscle performance of a mouse model of spinal cord injury [39]. Several molecular pathways are presumed to be involved in the myofiber phenotype shift from slow to fast, including the Ca2+ calcineurin nuclear factor’s activation of the T-cell 1 (NFATc1) pathway and the Ca2+ calmodulin kinase-histone deacetylase (HDAC) 4/5 myocyte enhancement factor 2 pathway [36, 40, 41]. Additionally, pathways involving extracellular-related kinase (ERK) 1/2, AMP-activated protein kinase (AMPK) sirtuin 1 peroxisome proliferator-activated receptor gamma coactivator 1-α (PGC1-α), and muscle-specific microRNAs are also related to myofiber type changes occurring with muscle disuse [36, 41]. However, no data are currently available about the relationship between CTE administration and myofiber type shift. Further experiments should seek to clarify the molecular mechanisms underlying the effects of CTE, as well as acteoside, on myofiber phenotype shifts and improvements in muscle performance; for this purpose, experiments on myoblast cell lines, such as C2C12 cells, should be performed [42].

Several other experiments should also be performed in future investigations of this subject. In order to robustly determine the effect of CTE on the myofiber type shift, CTE should be administered to various animal models of disuse muscle atrophy and tested, for example, on the diaphragm of

Cistanche extract has many benefits

mechanically ventilated animal [43] or on rats exposed to hypogravity by spaceflight [44]. Human data, such as in muscle atrophy due to bed rest, would also be necessary. Durations of CTE administration longer than 13 days should be tested, as they might lead to superior outcomes. Moreover, CTE administration after inducing muscle atrophy may also have an effect on the recovery phase of muscle atrophy, while our data are focused only on its preventive effect.

5. Conclusions

Our results indicate that CTE attenuates the decrease in MyHC IId/IIx expression and improves the performance of the atrophied muscle caused by disuse, without increasing muscle mass. Furthermore, they demonstrate that the novel casting method for hindlimb immobilization introduced here is both effective and useful in muscle wasting experiments. We found that it induced a reduction in muscle size, muscle CSA, and MyHC IId/IIx content, along with impaired muscle performance.

Data Availability

The data used to support the findings of this study are included in this article.

Conflicts of Interest

There are no conflicts of interest.

Authors’ Contributions

YK and CT conducted and designed the experiments. YK performed the experiments. YK and CT analyzed the data. YK, CT, YS, and TK wrote the paper. All authors approved the final version of the manuscript.

Acknowledgments

The authors are grateful to their colleagues in the Department of Japanese Oriental Medicine and the Division of Neuromedical Science for their generous support and productive discussions. The authors would like to thank Editage (http://www.editage.jp) for providing English language editing. This research was supported by a JSPS KAKENHI Grant, 2014, (grant number JP17H03558), a 2015 grant-in-aid for a cooperative research project from the Institute of Natural Medicine of the University of Toyama, and discretionary funds from the President of the University of Toyama in 2014, 2015, 2016, and 2017.

cistanche deserticola benefits

References

[1] N. Miljkovic, J.-Y. Lim, I. Miljkovic, and W. R. Frontera, “Aging of skeletal muscle fibers,” Annals of Rehabilitation Medicine, vol. 39, no. 2, pp. 155–162, 2015.

[2] Z. Aversa, P. Costello, and M. Muscaritoli, “Cancer-induced muscle wasting: latest findings in prevention and treatment,” Therapeutic Advances in Medical Oncology, vol. 9, no. 5, pp. 369–382, 2017.

[3] O. Schakman, S. Kalista, C. Barbe´, A. Loumaye, and J. P. Thissen, “Glucocorticoid-induced skeletal muscle atrophy,” Re International Journal of Biochemistry & Cell Biology, vol. 45, no. 10, pp. 2163–2172, 2013.

[4] S. S. Rudrappa, D. J. Wilkinson, P. L. Greenhaff, et al., “Human skeletal muscle disuse atrophy: effects on muscle protein synthesis, breakdown, and insulin resistance-a qualitative review,” Frontiers in Physiology, vol. 7, p. 361, 2016.

[5] R. G. Brower, “Consequences of bed rest,” Critical Care Medicine, vol. 37, no. 10, pp. S422–S428, 2009.

[6] K. Yamauchi, A. Yoshiko, S. Suzuki, et al., “Muscle atrophy and recovery of individual thigh muscles as measured by magnetic resonance imaging scan during treatment with the cast for ankle or foot fracture,” Journal of Orthopaedic Surgery (Hong Kong), vol. 25, no. 3, Article ID 2309499017739765, 2017.

[7] V. Dutt, S. Gupta, R. Dabur, E. Injeti, and A. Mittal, “Skeletal muscle atrophy: potential therapeutic agents and their mechanisms of action,” Pharmacological Research, vol. 99, pp. 86–100, 2015.

[8] S. Ciciliot, A. C. Rossi, K. A. Dyar, B. Blaauw, and S. Schiaffino, “Muscle type and fiber type specificity in muscle wasting,” Re International Journal of Biochemistry & Cell Biology, vol. 45, no. 10, pp. 2191–2199, 2013.

[9] J.-F. Desaphy, S. Pierno, A. Liantonio et al., “Antioxidant treatment of hindlimb-unloaded mouse counteracts fiber type transition but not atrophy of disused muscles,” Pharmacological Research, vol. 61, no. 6, pp. 553–563, 2010.

[10] H. Z. Feng, X. Chen, M. H. Malek, and J. P. Jin, “Slow recovery of the impaired fatigue resistance in port unloading mouse soleus muscle corresponding to decreased mitochondrial function and a compensatory increase in type I slow fibers,” American Journal of Physiology, Cell Physiology, vol. 310, no. 1, pp. C27–C40, 2016.

[11] The Ministry of Health, Labour and Welfare, “Cistanche Herb” in the Japanese Pharmacopoeia Seventeenth Edition, The Ministry of Health, Labour and Welfare, 2016.

[12] The Chinese Pharmacopoeia commission, Pharmacopeia of the People’s Republic of China Ver. 2015, China Medical Science and Technology Press, Beijing, China, 2015.

[13] N. Wang, S. Ji, H. Zhang, S. Mei, L. Qiao, and X. Jin, “Herba cistanches: anti-aging,” Aging and Disease, vol. 8, no. 6, pp. 740–759, 2017.

[14] Z. Fu, X. Fan, X. Wang, and X. Gao, “Cistanches Herba: an overview of its chemistry, pharmacology, and pharmacokinetics property,” Journal of Ethnopharmacology, vol. 219, pp. 233–247, 2018.

[15] T. N. Frimel, F. Kapadia, G. S. Gaidosh, Y. Li, G. A. Walter, and K. Vandenborne, “A model of muscle atrophy using cast immobilization in mice,” Muscle & Nerve, vol. 32, no. 5, pp. 672–674, 2005.

[16] N. Kawanishi, R. Nozaki, H. Naito, and S. Machida, “TLR4- defective (C3H/HeJ) mice are not protected from cast im- mobilization-induced muscle atrophy,” Physiological Reports, vol. 5, no. 8, 2017.

[17] L. Madaro, P. Smeriglio, M. Molinaro, and M. Bouche´, “Unilateral immobilization: a simple model of limb atrophy in mice,” Basic Applied Myology, vol. 18, pp. 149–153, 2008.

[18] E. L. Coutinho, A. R. S. Gomes, C. N. França, and T. F. Salvini, “A new model for the immobilization of the rat hind limb,” Brazilian Journal of Medical and Biological Research, vol. 35, no. 11, pp. 1329–1332, 2002.

[19] M. A. S. Khan, N. Sahani, K. A. Neville, et al., “Nonsurgically induced disuse muscle atrophy and neuromuscular dysfunction upregulates alpha7 acetylcholine receptors,” Canadian Journal of Physiology and Pharmacology, vol. 92, no. 1, pp. 1–8, 2014.

[20] A. Onda, H. Kono, Q. Jiao, et al., “New mouse model of skeletal muscle atrophy using spiral wire immobilization,” Muscle & Nerve, vol. 54, no. 4, pp. 788–791, 2016.

[21] A. Mokhtarian, J. P. Lefaucheur, P. C. Even, and A. Sebille, “Hindlimb immobilization applied to 21-day-oldmdx mice prevents the occurrence of muscle degeneration,” Journal of Applied Physiology, vol. 86, no. 3, pp. 924–931, 1999.

[22] A. Z. Caron, G. Drouin, J. Desrosiers, F. Trendz, and G. Grenier, “A novel hindlimb immobilization procedure for studying skeletal muscle atrophy and recovery in mouse,” Journal of Applied Physiology, vol. 106, no. 6, pp. 2049–2059, 2009.

[23] P. Londhe and D. C. Guttridge, “Inflammation induced loss of skeletal muscle,” Bone, vol. 80, pp. 131–142, 2015.

[24] R. D. Prisby, B. J. Behnke, M. R. Allen, and M. D. Delp, “Effects of skeletal unloading on the vasomotor properties of the rat femur principal nutrient artery,” Journal of Applied Physiology, vol. 118, no. 8, pp. 980–988, 2015.

[25] T. Ohira, F. Kawano, T. Ohira, K. Goto, and Y. Ohira, “Responses of skeletal muscles to gravitational unloading and/ or reloading,” Re Journal of Physiological Sciences, vol. 65, no. 4, pp. 293–310, 2015.

[26] J. W. Kim, S. K. Ku, K. Y. Kim, et al., “Schisandrae Fructus supplementation ameliorates sciatic neurectomy-induced muscle atrophy in mice,” Oxidative Medicine and Cellular Longevity, vol. 2015, Article ID 872428, 17 pages, 2015.

[27] B. A. Cisterna, C. Cardozo, and J. C. Sa´ez, “Neuronal involvement in muscular atrophy,” Frontiers in Cellular Neuroscience, vol. 8, p. 405, 2014.

[28] K. Sugimoto, “The application of lifestyle diseases-animal models to the research for sarcopenia,” Nippon Ronen Igakkai Zasshi. Japanese Journal of Geriatrics, vol. 50, no. 6, pp. 766–769, 2013.

[29] M. Tamaki, K. Miyashita, A. Hagiwara, et al., “Ghrelin treatment improves physical decline in sarcopenia model mice through muscular enhancement and mitochondrial activation,” Endocrine Journal, vol. 64, no. Suppl, pp. S47–S51, 2017.

[30] J. W. Kim, S.-K. Ku, M. H. Han, et al., “The administration of Fructus Schisandrae attenuates dexamethasone-induced muscle atrophy in mice,” International Journal of Molecular Medicine, vol. 36, no. 1, pp. 29–42, 2015.

[31] E. Ferraro, F. Pin, S. Gorini, et al., “Improvement of skeletal muscle performance in aging by the metabolic modulator Trimetazidine,” Journal of Cachexia, Sarcopenia and Muscle, vol. 7, no. 4, pp. 449–457, 2016.

[32] Y. Kishida, S. Kagawa, J. Arimitsu, et al., “Go-sha-jinki-Gan (GJG), a traditional Japanese herbal medicine, protects against sarcopenia in senescence-accelerated mice,” Phytomedicine, vol. 22, no. 1, pp. 16–22, 2015.

[33] R. M. Deacon, “Measuring motor coordination in mice,” Journal of Visualized Experiments: Jove, vol. 75, Article ID e2609, 2013.

[34] J. Talbot and L. Maves, “Skeletal muscle fiber type: using insights from muscle developmental biology to dissect targets for susceptibility and resistance to muscle disease,” Wiley Interdisciplinary Reviews: Developmental Biology, vol. 5, no. 4, pp. 518–534, 2016.

[35] F. M. Purves-Smith, N. Sgarioto, and R. T. Hepple, “Fiber typing in aging muscle,” Exercise and Sport Sciences Reviews, vol. 42, no. 2, pp. 45–52, 2014.

[36] D. J. Baker, D. Constantin-Teodosiu, S. W. Jones, J. A. Timmons, and P. L. Greenhaff, “Chronic treatment with the β2-adrenoceptor agonist prodrug BRL-47672 impairs rat skeletal muscle function by inducing a comprehensive shift to a faster muscle phenotype,” Journal of Pharmacology and Experimental Therapeutics, vol. 319, no. 1, pp. 439–446, 2006.

[37] A. Kodani, T. Kikuchi, and C. Tohda, “Acteoside improve muscle atrophy and motor function by inducing new myokine secretion in chronic spinal cord injury,” Journal of Neurotrauma, vol. 36, no. 12, pp. 1935–1948, 2019.

[38] B. S. Shenkman, “From slow to fast: hypogravity-induced remodeling of muscle fiber myosin phenotype,” Acta Naturae, vol. 8, no. 4, pp. 47–59, 2016.

[39] K. M. Baldwin, F. Haddad, C. E. Pandorf, R. R. Roy, and V. R. Edgerton, “Alterations in muscle mass and contractile phenotype in response to unloading models: role of transcriptional/pre translational mechanisms,” Frontiers in Physiology, vol. 4, p. 284, 2013.

[40] X. Chen, Y. Guo, G. Jia, G. Liu, H. Zhao, and Z. Huang, “Arginine promotes skeletal muscle fiber type transformation from fast-twitch to slow-twitch via Sirt1/AMPK pathway,” Re Journal of Nutritional Biochemistry, vol. 61, pp. 155–162, 2018.

[41] M. J. Tobin, F. Laghi, and A. Jubran, “Narrative review: ventilator-induced respiratory muscle weakness,” Annals of Internal Medicine, vol. 153, no. 4, pp. 240–245, 2010.

[42] D. A. Riley, S. Ellis, G. R. Slocom, T. Satyanarayana, J. L. W. Bain, and F. R. Sedlak, “Hypogravity-induced atrophy of rat soleus and extensor digitorum longus muscles,” Muscle & Nerve, vol. 10, no. 6, pp. 560–568, 1987.