Abstract: Objective: Apios americana, a plant used as a staple ingredient of native American diets, has various properties, including anti-cancer, anti-hyperglycemic, hypotensive, and anti-inflammatory activity. In Japan, Apios is used as a post-natal medication. After parturition, women undergo a period of recovery as they return to pre-pregnancy conditions. However, few health products that aid post-partum recovery are on the market. We explored whether Apios can accelerate the post-partum recovery process, in particular the involution of the uterus. Methods: Female rats kept in individual cages were mated with two male rats, with the exception of the control group (female rats without mating, on basal diet; n=6). After delivery, rats were divided into five groups based on their diet: basal diet (model; n=6); basal diet+oral intake at 5.4 g/kg of Chanfukang granules (a Chinese patent medicine preparation for post-partum lochia) (positive; n=6); basal diet containing 10% Apios powder (low; n=6); basal diet containing 20% Apios powder (medium; n=6); basal diet containing 40% Apios powder (high; n=6). Five days later, uteri and spleens were weighed. Uterus and spleen indices for each rat were calculated by dividing visceral weight by the total weight. Hormone and cytokine concentrations were measured using enzyme-linked immunosorbent assay (ELISA). Histological analysis of uteri was completed using hematoxylin and eosin (H&E) staining. Expression of matrix metalloproteinases and inhibitors in uteri was measured by western blotting. Results: Our results showed that Apios treatment reduced the post-partum uterus index and regulated the hormone concentrations. Moreover, we found that the process of uterine involution was accelerated, based on morphological changes in the uterus. In addition, our results indicated that Apios alleviated the inflammatory response induced by the involution process. Transforming growth factor β was also found to be regulated by Apios. There were significant downregulation of matrix metalloproteinases and upregulation of their inhibitors by Apios, which suggested that Apios increased the rate of the collagen clearance process. Conclusions: These results, based on experimental observations at the molecular and protein levels, verified our hypothesis that Apios can improve uterine involution, and demonstrated the potential application of Apios in post-partum care.

美国豆芋加快产后大鼠的子宫修复

[1]Alan E, Liman N, 2012. Immunohistochemical localization of beta defensins in the endometrium of rat uterus during the postpartum involution period. Vet Res Commun, 36(3):173-185.

[2]Al-Bassam MA, Thomson RG, O'Donnell L, 1981. Normal postpartum involution of the uterus in the dog. Can J Comp Med, 45(3):217-232.

[3]Alkema L, Chou D, Hogan D, et al., 2016. Global, regional, and national levels and trends in maternal mortality between 1990 and 2015, with scenario-based projections to 2030: a systematic analysis by the UN maternal mortality estimation inter-agency group. Lancet, 387(10017):462-474.

[4]Azawi OI, 2008. Postpartum uterine infection in cattle. Anim Reprod Sci, 105(3-4):187-208.

[5]Chitisankul WT, Shimada K, Omizu Y, et al., 2015. Mechanism of DDMP-saponin degradation and maltol production in soymilk preparation. LWT-Food Sci Technol, 64(1):197-204.

[6]Cui J, Kim Y, Lee EB, et al., 2016. Immunostimulatory activity of Apios tuber extract on RAW264.7 macrophage cells. J Bacteriol Virol, 46(4):248-257.

[7]Curry TE Jr, Osteen KG, 2003. The matrix metalloproteinase system: changes, regulation, and impact throughout the ovarian and uterine reproductive cycle. Endocr Rev, 24(4):428-465.

[8]Elliott L, McMahon KJ, Gier HT, et al., 1968. Uterus of the cow after parturition: bacterial content. Am J Vet Res, 29(1):77-81.

[9]Ferenczy A, Bertrand G, Gelfand MM, 1979. Studies on the cytodynamics of human endometrial regeneration: III. In vitro short-term incubation historadioautography. Am J Obstet Gynecol, 134(3):297-304.

[10]Gaide Chevronnay HP, Selvais C, Emonard H, et al., 2012. Regulation of matrix metalloproteinases activity studied in human endometrium as a paradigm of cyclic tissue breakdown and regeneration. Biochim Biophys Acta (BBA) Proteins Proteom, 1824(1):146-156.

[11]Galvão KN, Santos NR, Galvão JS, et al., 2011. Association between endometritis and endometrial cytokine expression in postpartum Holstein cows. Theriogenology, 76(2):290-299.

[12]Gray CA, Bartol FF, Tarleton BJ, et al., 2001. Developmental biology of uterine glands. Biol Reprod, 65(5):1311-1323.

[13]Griffin JF, Hartigan PJ, Nunn WR, 1974. Non-specific uterine infection and bovine fertility: I. Infection patterns and endometritis during the first seven weeks post-partum. Theriogenology, 1(3):91-106.

[14]Grota LJ, Eik-Nes KB, 1967. Plasma progesterone concentrations during pregnancy and lactation in the rat. J Reprod Fertil, 13(1):83-91.

[15]Harianja WY, Setiani O, Umaroh U, et al., 2017. The impact of pineapple (Ananas comosus (L.) Merr.) juice on fundal height in primigravida mothers during postpartum period. Belitung Nurs J, 3(2):134-141.

[16]https://doi.org/10.33546/bnj.76

[17]Henell F, Ericsson JLE, Glaumann H, 1983. An electron microscopic study of the post-partum involution of the rat uterus. With a note on apparent crinophagy of collagen. Virchows Arch B Cell Pathol Incl Mol Pathol, 42(1):271-287.

[18]Hoshikawa K, Juliarni, 1995. The growth of apios (Apios americana Medikus), a new crop, under field conditions. Jpn J Crop Sci, 64(2):323-327.

[19]Hulboy DL, Rudolph LA, Matrisian LM, 1997. Matrix metalloproteinases as mediators of reproductive function. Mol Hum Reprod, 3(1):27-45.

[20]Kasimanickam RK, Kasimanickam VR, Olsen JR, et al., 2013. Associations among serum pro- and anti-inflammatory cytokines, metabolic mediators, body condition, and uterine disease in postpartum dairy cows. Reprod Biol Endocrinol, 11:103.

[21]Kawamura J, Miura E, Kawakishi K, et al., 2015. Investigation of the safety and antihyperglycemic effect of Apios americana flower intake as a food material in normal and diabetic mice. Food Sci Technol Res, 21(3):453-462.

[22]Kikuta C, Sugimoto Y, Konishi Y, et al., 2012. Physicochemical and structural properties of starch isolated from Apios americana Medikus. J Appl Glycosci, 59(1):21-30.

[23]Kinugasa H, Watanbe Y, 1992. Nutritional composition of the tubers of American groundnut (Apios americana Medikus). Sonoda Women’s Coll Stud, 26:209-218.

[24]Krishnan HB, 1998. Identification of genistein, an anticarcinogenic compound, in the edible tubers of the American groundnut (Apios americana Medikus). Crop Sci, 38(4):1052-1056.

[25]Kuramoto S, Kaneyoshi G, Morinaga Y, et al., 2013. Angiotensin-converting enzyme-inhibitory peptides isolated from pepsin hydrolyzate of Apios americana tuber and their hypotensive effects in spontaneously hypertensive rats. Food Sci Technol Res, 19(3):399-407.

[26]LeBlanc MM, Hansen PJ, Buhi WC, 1988. Uterine protein secretion in postpartum and cyclic mares. Theriogenology, 29(6):1303-1316.

[27]Lin RY, Sullivan KM, Argenta PA, et al., 1995. Exogenous transforming growth factor-beta amplifies its own expression and induces scar formation in a model of human fetal skin repair. Ann Surg, 222(2):146-154.

[28]Manase K, Endo T, Chida M, et al., 2006. Coordinated elevation of membrane type 1-matrix metalloproteinase and matrix metalloproteinase-2 expression in rat uterus during postpartum involution. Reprod Biol Endocrinol, 4:32.

[29]Morales MR, Foster JG, 2009. Forage potential of American potato bean. Proceedings of the American Forage and Grassland Council. 2009 Annual Conference, Grand Rapids, Michigan.

[30]Murphy G, Nagase H, 2008. Progress in matrix metalloproteinase research. Mol Aspects Med, 29(5):290-308.

[31]Nagase H, Visse R, Murphy G, 2006. Structure and function of matrix metalloproteinases and TIMPs. Cardiovasc Res, 69(3):562-573.

[32]Nara K, Nihei KI, Ogasawara Y, et al., 2011. Novel isoflavone diglycoside in groundnut (Apios americana Medik). Food Chem, 124(3):703-710.

[33]Nguyen TTT, Shynlova O, Lye SJ, 2016. Matrix metalloproteinase expression in the rat myometrium during pregnancy, term labor, and postpartum. Biol Reprod, 95(1):24.

[34]O'kane S, Ferguson MWJ, 1997. Transforming growth factor βs and wound healing. Int J Biochem Cell Biol, 29(1):63-78.

[35]Petrovitch I, Jeffrey RB, Heerema-McKenney A, 2009. Subinvolution of the placental site. J Ultrasound Med, 28(8):1115-1119.

[36]Reynolds B, Blackmon W, Wickremesinhe E, et al., 1988. Domestication of Apios americana. In: Janick J (Ed.), Advances in New Crops: Proceedings of the First National Symposium New Crops: Research, Development, Economics, Timber Press, p.436-442.

[37]Rosen T, 2008. Placenta accreta and cesarean scar pregnancy: overlooked costs of the rising cesarean section rate. Clin Perinatol, 35(3):519-529.

[38]Salamonsen LA, 2003. Tissue injury and repair in the female human reproductive tract. Reproduction, 125(3):301-311.

[39]Sellers A, Woessner JF Jr, 1980. The extraction of a neutral metalloproteinase from the involuting rat uterus, and its action on cartilage proteoglycan. Biochem J, 189(3):521-531.

[40]Sheldon IM, 2004. The postpartum uterus. Vet Clin North Am Food Anim Pract, 20(3):569-591.

[41]Sheldon IM, Noakes DE, Rycroft AN, et al., 2002. Influence of uterine bacterial contamination after parturition on ovarian dominant follicle selection and follicle growth and function in cattle. Reproduction, 123(6):837-845.

[42]Shimizu K, Furuya T, Takeo Y, et al., 1983. Clearance of materials from breakdown of uterine collagen in mice during postpartum involution. Acta Anat (Basel), 116(1):10-13.

[43]Shkurupiy VA, Obedinskaya KS, Nadeev AP, 2011. Morphological study of the main mechanisms of myometrium involution after repeated pregnancies in mice. Bull Exp Biol Med, 150(3):378-382.

[44]Soo C, Beanes SR, Hu FY, et al., 2003. Ontogenetic transition in fetal wound transforming growth factor-β regulation correlates with collagen organization. Am J Pathol, 163(6):2459-2476.

[45]Takamoto N, Leppert PC, Yu SY, 1998. Cell death and proliferation and its relation to collagen degradation in uterine involution of rat. Connec Tissue Res, 37(3-4):163-175.

[46]Takashima M, Nara K, Niki E, et al., 2013. Evaluation of biological activities of a groundnut (Apios americana Medik) extract containing a novel isoflavone. Food Chem, 138(1):298-305.

[47]Umenishi F, Umeda M, Miyazaki K, 1991. Efficient purification of TIMP-2 from culture medium conditioned by human hepatoma cell line, and its inhibitory effects on metalloproteinases and in vitro tumor invasion. J Biochem, 110(2):189-195.

[48]Wang QH, Zhang S, Qin LM, et al., 2017. Yimu San improves obstetric ability of pregnant mice by increasing serum oxytocin levels and connexin 43 expression in uterine smooth muscle. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 18(11):986-993.

[49]WHO (World Health Organization), 2015. Childbirth: labour, delivery and immediate postpartum care. In: WHO, United Nations Population Fund, UNICEF (Eds.), Pregnancy, Childbirth, Postpartum and Newborn Care: A Guide for Essential Practice, 3rd Ed. World Health Organization, Geneva, p.85-88.

[50]WHO, 2018. Maternal mortality. http://www.who.int/en/news-room/fact-sheets/detail/maternal-mortality [Accessed on June 4, 2018].

[51]Willenbrock F, Murphy G, 1994. Structure-function relationships in the tissue inhibitors of metalloproteinases. Am J Respir Crit Care Med, 150(6 Pt 2):S165-S170.

[52]Wilson PW, Gorny JR, Blackmon WJ, et al., 1986. Fatty acids in the American groundnut (Apios americana). J Food Sci, 51(5):1387-1388.

[53]Woessner JF Jr, 1979. Total, latent and active collagenase during the course of post-partum involution of the rat uterus. Effect of oestradiol. Biochem J, 180(1):95-102.

[54]Woessner JF Jr, 1996. Regulation of matrilysin in the rat uterus. Biochem Cell Biol, 74(6):777-784.

[55]Wolf K, Sandner P, Kurtz A, et al., 1996. Messenger ribonucleic acid levels of collagenase (MMP-13) and matrilysin (MMP-7) in virgin, pregnant, and postpartum uterus and cervix of rat. Endocrinology, 137(12):5429-5434.

[56]Yan FJ, Yang YY, Yu LS, et al., 2017. Effects of C-glycosides from Apios americana leaves against oxidative stress during hyperglycemia through regulating mitogen-activated protein kinases and nuclear factor erythroid 2-related factor 2. J Agric Food Chem, 65(34):7457-7466.

[57]Yoshii A, Kitahara S, Ueta H, et al., 2014. Role of uterine contraction in regeneration of the murine postpartum endometrium. Biol Reprod, 91(2):32.

[58]Zhang YZ, Zhou CS, Tang SM, et al., 2011. Effect of AATI, a bowman-birk type inhibitor from Apios americana, on proliferation of cancer cell lines. Food Chem, 128(4):909-915.