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/lp/springer-journals/baofukang-suppository-promotes-the-repair-of-vaginal-epithelial-cells-ikC2m7gtdm
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- Springer Journals
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- Copyright © 2016 by The Author(s)
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- Life Sciences; Microbiology; Microbial Genetics and Genomics; Biotechnology
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- 2191-0855
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- 10.1186/s13568-016-0281-1
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Abstract
Vulvovaginal candidiasis ( VVC) is an opportunistic fungal infection predominantly caused by Candida albicans affect - ing a significant number of women of reproductive age. The Chinese medicine, the Baofukang suppository is widely used in the clinic for its antimicrobial activity and is therefore of great interest as a potential antifungal drug for the prevention of VVC. We evaluated the cytotoxic activity of the Baofukang suppository using the VK2/E6E7 vaginal epithelial cell ( VEC) line. When treated with the Baofukang suppository, all of the immunocompetent cytokines and chemokines (e.g., IL-2, IL-4, IL-6, IL-8, and IL-17) by infected VK2/E6E7 cells was statistically up-regulated (P < 0.05), except IL-4 (11.70 ± 1.82 vs. 14.88 ± 4.72, P = 0.343) compared to the infected control cells. The secretion of non-B IgG also exhibited the same trend. Our scanning electron microscopy results revealed that C. albicans can invade VECs by both induced endocytosis and active penetration. The Baofukang suppository could effectively inhibit the adhe - sion, hyphal formation, and proliferation, as well as notably restore the vaginal epithelial cell morphology, viability, and enhance the local immune function of the VECs. These preliminary results suggest promising antimicrobial properties of the Baofukang suppository, which may be efficacious as an antifungal therapy candidate via up-regulating Th1 cellular immunity, the Th17-axis of the innate immune response, and the secretion of vaginal epithelial-derived IgG. These combined effects collectively restore the immune function of the infected VECs against C. albicans in vitro. Keywords: Vulvovaginal candidiasis, Vaginal epithelial cells, Candida albicans, Baofukang suppository, Cytokines by published books and articles (Marti and Hine 1995; Introduction Newall et al. 1996). Baofukang suppository is a type of tra- The majority of women experience at least one episode ditional Chinese medicine that consists primarily of two of vulvovaginal candidiasis (VVC) in their lifetime (Sobel Chinese Medicinal Herbs: (1) zedoary turmeric oil; and et al. 1998). The vaginal epithelium as a mucosal surface (2) borneol. Zedoary turmeric oil is a volatile oil steam- is of immense importance in host defense and immune distilled from Curcuma phaeocaulis which contains a surveillance (Moyes et al. 2011). Specifically, it functions variety of effective components, including beta-elemene, as the first line of host defense against pathogen invasion curcumin, curzerenone, and Zimmer ketones currently to provide a physical barrier and protect underlying tis- in clinical use. These components may provide potent sues and organs (Cole 2006). pharmacological activities, such as antitumor or antivi- Products containing essential oils have been formu- ral effects, enhancing the immune response, particularly lated for intra-vaginal use, and are often recommended by promoting the regeneration and repair of damaged or as home remedies for the treatment of vaginal candidiasis inflamed mucosa (Kamazeri et al. 2012). Due to its potent pharmacological action, this plant extract has begun to *Correspondence: 17301255426@163.com attract some significant attention. Borneol is extracted Ting Li and Xiaoxi Niu contributed equally to this work as joint first from the essential oil of various subtropical and ever- author Department of Obstetrics and Gynecology, Peking University First green broad-leaved such as Cinnamomum and Laura- Hospital, 8 Xishiku Street, Xicheng District, Beijing 100034, China ceae, and a bicyclic monoterpenoid alcohol commonly Full list of author information is available at the end of the article © The Author(s) 2016. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. Li et al. AMB Expr (2016) 6:109 Page 2 of 8 used in traditional Chinese medicine as the adjuvant for at a concentration of 5, 10, 20, 40, 80, and 160 µg/mL. A more than 1500 years (Jiang et al. 2008). It demonstrates total volume of 200 µL VK2 cell suspension was seeded anti-inflammatory, analgesic, and antibacterial properties, into each well of a 96-well microtiter plate and placed while also accelerating percutaneous drug absorption and into a humidified atmosphere containing 5% CO at 37 °C increasing the bioavailability of drugs in the brain tissue for 24 h before the cells were treated. Untreated cells that (Almeida et al. 2013; Slamenova et al. 2009). Therefore, received only media were used as the negative control. the aims of this work was to evaluate the in vitro antifun- Concentrations of 0, 5, 10, 20, 40, 80, and 160 µg/mL Bao- gal properties and mechanisms of the Baofukang supposi- fukang suppository were added to the VK2 cells for 24 h. tory, which would be further applied widely in clinic as a The cells in each well were incubated in 100 µL K-SFM potential antifungal drug for the prevention of VVC. containing 10 µL CCK-8 reagents at 37 °C for 1 h. The plate was then shaken on an automatic mixer for 3 min and the Materials and methods absorbance at 450 nm (A ) was measured using a Multi- Vaginal epithelial cell culture scan GO micro-plate reader. The results were expressed as The VK2/E6E7 vaginal epithelial cell line (ATCC CRL- the percentage of cell viability and plotted. 2616), obtained from the American Type Culture Collec- Cell viability (%) = [A (treated) − A (blank)]/ 450 450 tion (ATCC; Rock ville, MD, USA) is an epithelial cell line [ A (control) − A (blank)] × 100% 450 450 derived from the vaginal mucosa of a healthy premeno- pausal female undergoing vaginal repair surgery, that The concentration of the sample that inhibited 10% cell was subsequently immortalized with human papilloma- growth as calculated by SPSS 13.0 was the 10% inhibition virus 16/E6E7. VK2 cells were cultured in keratinocyte- concentration (IC ). This dose was defined as a safe dose serum free medium (K-SFM, Gibco, USA) supplemented with little poisonous side effects—the highest concentra - with 5 ng/mL recombinant epidermal growth factor and tion where still no effect of the Baofukang suppository 50 µg/mL bovine pituitary extract (Invitrogen Corpora- on cell viability (≥90% survival) (Namiecinski et al. 2004; tion, Grand Island, NY, USA), 100 U/mL penicillin (Life Qiao et al. 2013). Technologies, Grand Island, NY, USA), 100 µg/mL strep- tomycin (Life Technologies) and 0.4 mM CaCl at 37 °C Cytokine and chemokine analysis of coculture with 5% CO and a high humidity environment. A sub- supernatants culture of the cells was performed every 3–4 days. For the examination of cytokines and chemokines, epi- thelial cells (1 × 10 cells/mL) were cocultured with C. Microbial strains and growth conditions candida (1 × 10 /mL) at a ratio of 1:1 in separate wells Candida albicans collection strains (ATCC-11006) were for 12 h for the VK2 cell line cells in a total volume of grown aerobically overnight on Sabouraud-dextrose agar 2 mL K-SFM complete medium in 24-well tissue culture plates (Becton Dickinson, Cockeysville, MD, USA) at plates (Costar, Corning, NY, USA). Following a coculture 37 °C until the mid-exponential growth phase. The blas - for 12 h, the culture medium was aspirated, washed three toconidia were collected and resuspended in RPMI 1640 times with PBS, and replaced with 1 mL different con - and adjusted to 1.0 × 10 cells/mL after counting with a centrations 20 µg/mL of Baofukang suppository (IC ) hemocytometer (Hausser Scientific; Horsham, PA, USA). as described above for additional 24 h. The supernatants were collected and centrifuged at 12,000g for 5 min and Drug preparation finally stored at − 80 °C until an enzyme-linked immuno- Baofukang suppository (Hainan bikai Pharmaceuti- sorbent assay (ELISA, eBioscience, USA) was performed. cal Co., Ltd.) is a traditional Chinese Medicine, with The supernatants were assayed for the levels of IL-6, IL-2, every 1.74 g tablet consisting of 88 mg zedoary turmeric IL-4, IL-8, and IL-17 cytokines according to the manufac- oil, 75 mg borneol, and other components as a preser- turer’s instructions. New standard curves were generated vation matrix. One vaginal suppository tablet (water for every set of experiments. The absorbance values and soluble) was dissolved in 44 mL serum-free RPMI1640 concentrations of each cytokine were determined using a culture medium to prepare a drug stock solution of Ceres 900 automated microplate reader (Bio-Tek Corp., 3.95 × 10 µg/mL, and was passed through a 0.22 µM Wisnooski, VT, USA) and Kineticalc software (Bio-Tek). membrane filter for sterilization. All drug solutions were Each independent experiment was performed in triplicate. stored at −20 °C until further experiments. Epithelial‑derived IgG and sIgA analysis of coculture Evaluation of cytotoxic activity supernatants CCK-8 (Dojindo Laboratories, Tokyo, Japan) was used to To further explore the local immune function of vaginal evaluate the cytotoxicity of the Baofukang suppository epithelial cells, we stimulated VK2/E6E7 with C. candida Li et al. AMB Expr (2016) 6:109 Page 3 of 8 (1 × 10 /mL) and detected the level of secreted non-B IgG and IgA in the culture supernatants (collected as described above) by an ELISA (eBioscience). The ELISAs were conducted as mentioned above. Scanning electron microscopy (SEM) Specimens were fixed overnight in 2.5% glutaraldehyde in 0.1 M sodium cacodylate buffer (pH 7.4; Electron Microscopy Sciences, Hatfield, PA, USA) at 4 °C, rinsed three times with PBS, dehydrated in graded ethanol (25, 50, 75, 95, and 100%), and dried using the critical point drying method (BALTEC, Balzers, Liechtenstein). The dried samples were glued onto SEM stubs, sputter-coated with a 10 nm thick layer of gold (BALTEC, Balzers, Fig. 1 Cytotoxicity of VECs in the presence of the Baofukang sup- Liechtenstein), and examined using a scanning electron pository. The vaginal epithelial cells were treated with the Baofukang microscope (S-3400 N, Hitachi, Japan). suppository for 24 h. The mean values for the remaining four values and standard deviations (error bars) are shown Statistical analysis All data are presented as the mean ± standard devia- tion of three independent measurements. The statistical (45.13 ± 4.718 pg/mL, P = 0.526). Comparatively, the analyses were conducted using SPSS version 13.0 (SPSS, Baofukang suppository stimulated a significant up-reg - Chicago, IL, USA). A statistical comparison between the ulation in the production of IL-8 (25.91 ± 0.50 pg/mL, groups was carried out using a one-way ANOVA. Sub- P < 0.0001) and a significant down-regulation in the pro - sequent comparisons were performed using the LSD duction of IL-4 (25.06 ± 1.65 pg/mL, P = 0.018) and IL-6 method. Significant differences were defined as having a (23.08 ± 0.12 pg/mL, P < 0.0001) by the VK2/E6E7 cells P value of less than 0.05. was observed (Fig. 2). After 12 h of challenge with C. albicans, IL-2, IL-4, Results IL-6, IL-8, and IL-17 had substantially declined (0.4- to Eec ff ts of the Baofukang suppository on VK2/E6E7 cell 0.8-fold, P < 0.05; Fig. 2); however, when treated with viability 20 µg/mL Baofukang suppository, all of the above-men- To mimic clinical situations in which antibiotics or anti- tioned cytokines in the infected VK2/E6E7 cell cultures fungals may be safe and well-tolerated in the human were significantly up-regulated (IL-2: 25.14 ± 3.43 vs. body, we determined whether the Baofukang supposi- 31.59 ± 1.90 pg/mL, P = 0.030; IL-4: 11.70 ± 1.82 vs. tory affects cell viability. The A450 of the VK2/E6E7 cells 14.88 ± 4.72 pg/mL, P = 0.343; IL-6: 8.91 ± 0.65 vs. treated with the Baofukang suppository at doses of 0, 5, 13.74 ± 0.51 pg/mL, P < 0.0001; IL-8: 12.41 ± 0.06 vs. 10, 20, 40, 80, and 160 µg/mL were plotted in Fig. 1. As 17.63 ± 0.41 pg/mL, P < 0.0001; IL-17: 12.99 ± 2.57 vs. shown in Fig. 1, high doses (>20 µg/mL) of the Baofukang 19.52 ± 2.13 pg/mL, P = 0.039; respectively), when com- suppository did inhibit vaginal epithelial cell viability or pared to the untreated vaginal cells infected with C. albi- growth, while there were no observed changes in the cells cans (Fig. 2). We determined the Th1/Th2 balance by exposed to the low doses (≤20 µg/mL). The IC calcu- calculating the IL-2/IL-4 ratio (Table 1). lated by SPSS 13.0 was 19.89 µg/mL, and was considered to be a safe dose for VK2 cells with minimal toxic side The Baofukang suppository modulates epithelial‑derived effects, and was selected for further experiments. IgG secreted by human vaginal epithelial cells To determine whether infected, uninfected or treated Baofukang suppository modulates cytokine release vaginal epithelial cells secrete epithelial-derived IgG by human vaginal epithelial cells and sIgA, the culture supernatants were examined by an In the present study, at baseline, the level of IL-2, IL-4, ELISA (Fig. 3). Secretory IgA (sIgA) that we previously IL-6, IL-8, and IL-17 production by VK2 cells were assumed to be the most abundant Ig isotype secreted in 46.81 ± 3.07, 34.42 ± 5.60, 27.96 ± 0.15, 21.53 ± 0.78, culture supernatants was undetectable in this experiment and 43.36 ± 3.68 pg/mL, respectively (Fig. 2). After 24 h (data not shown). of co-incubation with 20 µg/mL of the Baofukang sup- Surprisingly, we found that the VECs spontaneously pository alone, there were no significant changes in the secrete epithelial-derived IgG under baseline conditions levels of IL-2 (45.76 ± 3.31 pg/mL, P = 0.679) or IL-17 Li et al. AMB Expr (2016) 6:109 Page 4 of 8 Fig. 2 The production of IL-2, IL-4, IL-6, IL-8, and IL-17 (expressed as pg/mL) by the vaginal epithelial cell line, VK2/E6E7 cells cultivated alone, grown with 20 µg/mL the Baofukang suppository, and infected with C. albicans (1 × 10 /mL). The cytokine levels were measured after 12 h of infection and subsequent 24 h of co-incubation with 20 µg/mL of the Baofukang suppository. V represents the vaginal epithelial cells cultivated alone; V+B rep- resents vaginal epithelial cells co-incubated with 20 µg/mL of the Baofukang suppository for 24 h; V+C represents vaginal epithelial cells infected with C. albicans for 12 h; V+C+F50 represents the vaginal epithelial cells infected with C. albicans for 12 h, then treated with 20 µg/mL of Baofukang suppository for another 24 h Table 1 Th1/Th2 cytokines/ratios Cytokine (pg/ml) V V+B V+C V+C+B F value P value IL-2 46.81 ± 3.07 45.76 ± 3.31 25.14 ± 3.43 31.58 ± 1.90 38.291 <0.0001* IL-4 37.65 ± 0.85 25.70 ± 0.60 13.12 ± 0.76 16.57 ± 4.99 54.464 <0.0001* IL2/IL-4 1.24 ± 0.08 1.78 ± 0.17 1.93 ± 0.33 2.03 ± 0.63 2.684 0.117 V represents the vaginal epithelial cells cultivated alone; V+B represents the vaginal epithelial cells co-incubated with 20 µg/mL of the Baofukang suppository for 24 h; V+C represents the vaginal epithelial cells infected with C. albicans for 12 h; V+C+B represents the vaginal epithelial cells infected with C. albicans for 12 h, then treated with 20 µg/mL of the Baofukang suppository for another 24 h * P < 0.0001 (Fig. 3). The baseline level of IgG secreted by the VK2 The cell surface of the normal, uninfected cells was cells was 0.64 ± 0.13 µg/mL, which dropped sharply covered with microvilli or a microvilli crest that was an following infection or co-incubation with the Baofu- irregular, loose net-like membrane ruffle (Fig. 4A). At 6 kang suppository alone (P < 0.0001). However, when and 12 h post-infection, epithelial cell adherence and the infected VECs were treated with 20 µg/mL Baofu- invasion were observed (Fig. 4B, D). Our results suggest kang suppository, the interaction pattern of this sup- that C. albicans can invade vaginal epithelial cells by two pository was changed, and the level of IgG secreted by distinct mechanisms that ultimately result in cellular the treated VK2/E6E7 cells was statistically up-regulated damage. The first is the induction of cellular endocytosis by the C. albicans hypha, and the other is via active pen (0.42 ± 0.06 µg/mL) over the untreated infected cells - (0.24 ± 0.02 µg/mL) (P = 0.025). Despite this increase, etration without pseudopod formation and endocytosis. the levels did not reach the baseline values (P = 0.013). After 24 h of treatment with the Baofukang supposi- tory, as shown at ×3000 high magnification in Fig. 4D, Baofukang suppository promotes the repair of infected the invasive blastoconidia and hyphae initially observed cells were significantly reduced or completely absent. Treated For further insight into the interactions of C. albicans cells with a normal shape, good cell viability, relatively with VECs, VEC monolayers were preferred even though intact and smooth cell membrane, were well adhered to the vaginal mucosa is composed of stratified squamous the wall and completely stretched, similar to the unin- epithelium because only the superficial epithelial cells fected cells shown in Fig. 4A. that are exposed to the luminal surface interact with C. These results illustrate that the Baofukang suppository albicans. Thus, establishing models with multiple layers could not only effectively inhibit the adhesion, hyphal of cells could complicate the quantification of the interac - formation, and proliferation of C. albicans, but also tions between the epithelial cells and the pathogen. notably restore the VEC morphology and viability. u Th s, Li et al. AMB Expr (2016) 6:109 Page 5 of 8 To mimic the effect of the Baofukang suppository in treating vulvovaginal candidiasis under in vivo condi- tions, the VK2/E6E7 cells were infected with C. albi- cans for 12 h and then treated with the Baofukang suppository for 24 h. The levels of IL-2, IL-6, IL-8, and IL-17 were significantly increased, while IL-4 was only slightly increased in the treated cells compared with the infected control. Th1 cells are associated with inter - feron γ or L-2 cooperating with cytotoxic CD8+ T cells, while Th2 cells are associated with IL-4 or IL-6 promot - ing a humoral, proinflammatory response (Schilling et al. 2007; Woo et al. 2015). A increase in the balance of the Th1/Th2 ratio is associated with the enhancement of cell-mediated immunity. Thus, the Baofukang supposi - tory may enhance or restore a protective Th1 response in Fig. 3 The production of epithelial-derived IgG (expressed in µg/ mL) by the vaginal epithelial cell line, VK2/E6E7 cells. The IgG levels infected cells, which represents the immunological hall- were measured after 12 h of infection and the subsequent 24 h of mark of candidal lesions believed to play a crucial role co-incubation with the Baofukang suppository. V represents the in the clearance of mycotic infection (Fidel 2002). IL-2 vaginal epithelial cells cultivated alone; V+B represents the vaginal and IL-4 are the classically representative of Th1 and Th2 epithelial cells co-incubated with 20 µg/mL of teh Baofukang sup- cytokines, respectively (Fidel and Sobel 1994; Romani pository for 24 h; V+C represents the vaginal epithelial cells infected with C. albicans for 12 h; V+C+B represents the vaginal epithelial 1999; Woo et al. 2015). A strong role for Th1-type cell- cells infected with C. albicans for 12 h, then treated with 20 µg/mL of mediated immunity against Candida was demonstrated the Baofukang suppository for another 24 h. ***Significant difference by various experimental models (Klein et al. 1984; Clift ## compared to the V group (P < 0.0001), significant difference com- 1984). An increased Th1/Th2 ratio (IL-2/IL-4) is associ - pared to the V+C group (P = 0.025). Each sample was repeated three ated with the activation of cell-mediated immunity and times. The error bars indicate the standard deviation would be potentially beneficial for pathogen or cancer elimination, while a decreased ratio would raise the risk of inflammation progression. Our results indicate that this serves to enhance the local immune function of the the IL-2/IL-4 ratio was significantly increased in the Bao - VECs. fukang suppository-treated cells and indicates that the Baofukang suppository can indirectly up-regulate the Discussion vaginal local cellular immunity under the infective status VECs lining the mucosal surfaces of the vagina provide promoting the host defense against invading pathogenic a first-line barrier defense system for both innate and microorganisms. acquired immune functions. As in anticandidal mucosal Moreover, evidence has been mounting that a Th17- immunity, the innate role of the epithelial response driven immune response plays a predominant role in involves the release of epithelial cell-derived cytokines modulating a defensive mucosal immune response and chemokines in response to C. albicans appear to against Candida in both mice and humans (Huang et al. have important roles recruiting and activating a variety 2004; Conti et al. 2009; Eyerich et al. 2008). An impaired of immune cells, immunoregulation, and tissue repair IL-17 response appears to be responsible for the patho- (Dongari-Bagtzoglou et al. 2005; Schaller et al. 2004). This genesis of chronic mucocutaneous candidiasis (Eyer- serves to further mediate the innate and adaptive responses ich et al. 2008). Our data also affirm that the Baofukang of protective mucosal immunity against C. albicans. Our suppository partly restores IL-17-production by VECs results have demonstrated that VECs secrete various solu- against Candida infection in vitro, thus, initiating an early ble immunological mediators under normal culture condi- Th17-type innate immune response against extracellu - tions. However, after challenging the cells with C. albicans lar Candida adhesion and filamentation. Active cytokine for 12 h, the secretion of all the cytokines and chemokines production by VECs would enable a rapid response to produced by the VECs were significantly decreased. This an infection and provide protection within the vaginal decrease may partially be due to a direct invasion of C. albi- microenvironment. cans and a strongly diminished total number of VK2/E6E7, Immunoglobulins are one of the key molecules of the resulting in damaging the local innate immune function. humoral immune response, and they have previously Martinez et al. (2009) confirmed that the numbers of VK2/ thought to be produced only by B cells, and no other E6E7 cells remained stable within 6 h of C. albicans infec- cell types. However, 20 years ago, a series of studies tion, which decreased to 1/8-fold after inoculation for 12 h. Li et al. AMB Expr (2016) 6:109 Page 6 of 8 Fig. 4 Scanning electron micrographs (SEM) of the vaginal epithelial cells. SEM of the control cells (A), C. albicans infected cells at 6 h (B), C. albicans infected cells at 12 h (C) and treated cells (D), the latter represents the vaginal epithelial cells infected with C. albicans for 12 h, then treated with 20 µg/mL Baofukang suppository for another 24 h. Fusion of the microvilli-like structures forming membrane leaflets that envelop the invading hyphal cells is indicated by the red arrow; damaged cell debris is indicated by the white arrow; pseudohyphae elements “engulfed” into vaginal epithelial cells are indicated by yellow arrows; a budding spore is indicated by blue arrows demonstrated that non-B cancer cells and normal non-B can be expressed in VECs, and they appear to be involved cells (Qiu et al. 2003, 2013; Zhao et al. 2011) are capa- in the innate immune response of the vagina against ble of producing Igs, but it remains unclear whether nor- mycotic infections, which can be partially repaired by the mal VECs express functional Ig molecules. We cultured Baofukang suppository treatment. Further studies should the immortalized VEC lines, VK2 cells, and used an ascertain if vaginal epithelial-derived IgG participates in ELISA to determine whether IgG or sIgA was secreted the local mucosal immunity of the vagina against various by the VECs. To our surprise, sIgA was undetectable in common pathogens. cell supernatants. In this report, we first noted epithe - The SEM observations were first conducted to visualize lial-derived IgG were secreted by VECs in vitro, which the different stages (6 and 12 h) of the interaction of C. challenges the classical concept that B cells are the only albicans with VECs. Similar to the pathological processes source of Ig. Antibody-mediated protection containing of vulvovaginal candidiasis, the yeast and filamentous an anti-secreted aspartyl proteinase antibody or anti- forms of C. albicans are capable of adhering and invading Candida-mannoprotein appear to play an indispensable the VECs, enabling the fungal cells to translocate across role in mucosal immunity against vaginitis. However, this the vaginal mucosal barrier. C. albicans invade the vagi- hypothesis cannot explain the clinical phenomenon that nal epithelial monolayer via two mechanisms: (1) induced no antibody deficiency was observed in recurrent VVC endocytosis; or (2) active penetration (Zhu and Filler women and the absence of protective Candida-specific 2010; Dalle et al. 2010; Naglik et al. 2011). Endocytosis is antibody production in inoculated mice (Ashman et al. defined as the engulfment of epithelial cells with mem - 2004). Our study confirms our hypothesis that non-B IgG brane protrusions at the point of entry of the invading Li et al. AMB Expr (2016) 6:109 Page 7 of 8 Competing interests hypha, and active penetration is defined as hypha pen - The authors declare that they have no competing interests. etration on the epithelial cell surface at their apical side at the point of entry of an invading hypha (Dalle et al. 2010). Ethical approval This article does not contain any studies with human participants or animals The two mechanisms appear throughout the entire pro - performed by any of the authors. cess of invasion, while the former occurs more frequently during the early-stage and the latter more frequently in Funding This work was supported by a grant from the National Natural Science Foun- the late-stage in vitro (Dalle et al. 2010). We also confirm dation of China (Grant Number 81571394). that hypha formation is the key to fungal invasion and damage, and VK2 cells, when not fully damaged by fungal Received: 8 September 2016 Accepted: 31 October 2016 cells, can kill or “engulf ” the attached C. albicans. How- ever, this antifungal phenomenon was not observed when the VK2 cells were killed by C. albicans. When treated with a Baofukang suppository for 24 h References compared with an infectious condition, C. albicans adhe- Almeida JR, Souza GR, Silva JC, Saraiva SR, Júnior RG, Quintans Jde S, Barreto sion, invasion, and cellular injury could be restored to a Rde S, Bonjardim LR, Cavalcanti SC, Quintans LJ Jr (2013) Borneol, a bicy- clic monoterpene alcohol, reduces nociceptive behavior and inflamma- normal condition. The antimicrobial activity of the Bao - tory response in mice. Sci World J 2013:808460. doi:10.1155/2013/808460 fukang suppository of C. albicans could, in part, be asso- Ashman RB, Farah CS, Wanasaengsakul S, Hu Y, Pang G, Clancy RL (2004) Innate ciated with a major constituent of Zedoary turmeric oil versus adaptive immunity in Candida albicans infection. Immunol Cell Biol 82:196–204 6. Moreover, these results also demonstrated a direct role Clift RA (1984) Candidiasis in the transplant patient. Am J Med 77:34–38 for the Baofukang suppository in inhibiting germ tube Cole AM (2006) Innate host defense of human vaginal and cervical mucosae. formation and hyphal elongation for antifungal defense at Curr Top Microbiol Immunol 306:199–230 Conti HR, Shen F, Nayyar N, Stocum E, Sun JN, Lindemann MJ, Ho AW, Hai JH, mucosal surfaces. Therefore, it can be used as a natural Yu JJ, Jung JW, Filler SG, Masso-Welch P, Edgerton M, Gaffen SL (2009) preservative in food or pharmaceuticals. 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J Infect Dis 190:624–631 Abbreviations Jiang XF, Zou JL, Yuan YM, Francis CL, Qiao YJ, Yao MC (2008) Preliminary study: VVC: vulvovaginal candidiasis; ATCC: American Type Culture Collection; K-SFM: biotransformation of borneol to camphor in mice, rats, and rabbits. Mode keratinocyte-serum free medium; IC : the 10% inhibition concentration; Tradit Chin Med Mater Med 10:27–36 ELISA: enzyme-linked immunosorbent assay; SEM: scanning electron micros- Kamazeri TS, Samah OA, Taher M, Susanti D, Qaralleh H (2012) Antimicrobial copy; sIgA: secretory IgA; VECs: vaginal epithelial cells. activity and essential oils of Curcuma aeruginosa, Curcuma mangga, and Zingiber cassumunar from Malaysia. Asian Pac J Trop Med 5:202–209 Authors’ contributions Klein RS, Harris CA, Small CB, Moll B, Lesser M, Friedland GH (1984) Oral can- The main experimental conception and design: ZL; Performed the experi- didiasis in high-risk patients as the initial manifestation of the acquired ments: TL and XN; Analyzed the data and contributed reagents: TL, XN and ZL; immunodeficiency syndrome. N Engl J Med 311:354–358 Writing the manuscript: TL and XN. All authors read and approved the final Marti J, Hine A (1995) The alternative health and medicine encyclopedia. Gale manuscript. Research International Inc. New York Martinez RC, Seney SL, Summers KL, Nomizo A, De Martinis EC, Reid G (2009) Author details Eec ff t of Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 on Department of Obstetrics and Gynecology, Peking University First Hospital, 8 the ability of Candida albicans to infect cells and induce inflammation. Xishiku Street, Xicheng District, Beijing 100034, China. 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Journal
AMB Express – Springer Journals
Published: Nov 9, 2016