Wang F, Huo Y, Yin L R, Sun B, Zhang P P
Department of Gynecology, the Second Hospital of Tianjin Medical University, Tianjin 300211, China.
Zhonghua Fu Chan Ke Za Zhi. 2016 Jul 25;51(7):530-4. doi: 10.3760/cma.j.issn.0529-567X.2016.07.009.
To study the effects of antimicrobial peptide LL-37 expressed and purified from prokaryotes on candida albicans growth.
(1)Thirty female Kunming mice were treated with estrogen and white candida yeast suspension were poured into vagina to establish a vulvovaginal candidiasis(VVC)murine model. After successful establishing the VVC mouse model, mice were randomly sorted into test group(n=15)and control group(n=15). Suspension(30 μl, 100 μg/ml)of recombinant peptide LL-37 expressed and purified in Prokaryotes was given by intravaginal administration to the test group for 5 days, while the same amount of phosphate buffered saline(PBS)was given to the control group.(2)Tweenty-four hours after treatment, the fungal burden and colony-forming unit(CFU)of vaginal fluids were evaluated. All mice were subsequently sacrificed and vaginal tissues were harvested for tissue homogenate preparation. ELISA was used to determine the levels of nterleukin-10(IL-10)and interferon-γ(IFN-γ)in the isolated vaginal tissues.
(1)VVC mouse model was established successfully in this study. Vaginal mucosa congestion, edema, vaginal plica disappearing were obviously observed in the control group. After treatment with recombinant protein LL-37 vaginal mucosa has no obvious change in the test group.(2)Fungal burden and CFU of vaginal fluids were significantly lower in the test group [(4.8±1.0)×10(4) CFU/ml]than that in the control group [(8.5±2.1)×10(4) CFU/ml, P=0.017]. IFN-γ level of the test group was increased [(257±11)vs(197±4)pg/ml, P=0.000], while the level of IL-10 was reduced [(287 ± 15)vs(379 ± 17)pg/ml P=0.000] resulting in a the ratio of IFN-γ/IL-10 was in significantly higher in test group(0.892±0.008 vs 0.496±0.013, P=0.000).
Recombinant protein LL-37 expressed and purified from prokaryotes inhibits the growth candida albicans and improves vaginal immunity by adjusting IFN-γ and IL-10 secretion in the VVC mouse model, highlighting the therapeutic potential of LL-37 for VVC.
研究原核表达纯化的抗菌肽LL-37对白色念珠菌生长的影响。
(1)30只雌性昆明小鼠用雌激素处理后,将白色念珠菌酵母悬液注入阴道,建立外阴阴道念珠菌病(VVC)小鼠模型。成功建立VVC小鼠模型后,将小鼠随机分为试验组(n=15)和对照组(n=15)。试验组经阴道给予原核表达纯化的重组肽LL-37悬液(30 μl,100 μg/ml),连续5天,对照组给予等量的磷酸盐缓冲盐水(PBS)。(2)治疗24小时后,评估阴道分泌物中的真菌负荷和菌落形成单位(CFU)。随后处死所有小鼠,采集阴道组织制备组织匀浆。采用酶联免疫吸附测定法(ELISA)检测分离的阴道组织中白细胞介素-10(IL-10)和干扰素-γ(IFN-γ)的水平。
(1)本研究成功建立了VVC小鼠模型。对照组可见阴道黏膜充血、水肿,阴道皱襞消失。试验组用重组蛋白LL-37治疗后,阴道黏膜无明显变化。(2)试验组阴道分泌物中的真菌负荷和CFU[(4.8±1.0)×10⁴CFU/ml]明显低于对照组[(8.5±2.1)×10⁴CFU/ml,P=0.017]。试验组IFN-γ水平升高[(257±11)对(197±4)pg/ml,P=0.000],而IL-10水平降低[(287±15)对(379±17)pg/ml,P=0.000],导致试验组IFN-γ/IL-10比值显著升高(0.892±0.008对0.496±0.013,P=0.000)。
原核表达纯化的重组蛋白LL-37在VVC小鼠模型中通过调节IFN-γ和IL-10的分泌抑制白色念珠菌生长并提高阴道免疫力,突出了LL-37对VVC的治疗潜力。