Li Jin-Qing, Song Ya-Li, Xun Tian-Rong, Tan Sui-Yi, Liu Shu-Wen
School of Pharmaceutical Sciences, Southern Medical University, Guangzhou 510515, China.E-mail:
Nan Fang Yi Ke Da Xue Xue Bao. 2017 Jul 20;37(7):907-913. doi: 10.3969/j.issn.1673-4254.2017.07.09.
To investigate the inhibitory effect of lactic acid on semen-derived amyloid (SEVI) fibril formation.
PAP248-286 (2 mg/mL) was incubated with 4.0, 2.0, 1.0, 0.5, 0.25, and 0.125 mg/mL of lactic acid. After incubation for different times, aliquots were drawn from each sample for Thioflavin T (ThT) and Congo red staining to monitor semen-derived amyloid fibril formation. The β sheet structure formation of PAP248-286 was measured by circular dichroism spectrum, and the morphology of amyloid fibrils incubated with or without lactic acid was observed with transmission electron microscopy (TEM). The enhancing effect of amyloid fibril incubated with lactic acid at different time points was determined using virus infection assay. PAP248-286 (2 mg/mL) was incubated with dilutions of vaginal secretion from healthy women, and amyloid fibril formation was detected with ThT and Congo red staining.
Lactic acid inhibited SEVI fibril formation in a dose-dependent manner in vitro. Lactic acid at 0.5 mg/mL completely inhibited 2 mg/mL SEVI fibril formation within 48 h. After incubation for 48 h, lactic acid at 1 mg/mL inhibited the formation of β-sheet structure of SEVI (2 mg/mL) and completely inhibited 2 mg/mL PAP248-286 aggregation as observed with TEM. In the presence of lactic acid, PAP248-286 lost the ability to enhance virus infection. Vaginal secretion inhibited SEVI fibril formation in a dose-dependent manner, and virtually no SEVI fibril occurred after incubation of 2 mg/mL PAP248-286 with 67% vaginal secretion.
Lactic acid inhibits SEVI fibril formation in vitro.
研究乳酸对精液衍生淀粉样蛋白(SEVI)纤维形成的抑制作用。
将PAP248 - 286(2 mg/mL)与4.0、2.0、1.0、0.5、0.25和0.125 mg/mL的乳酸孵育。在不同时间孵育后,从每个样品中取出等分试样进行硫黄素T(ThT)和刚果红染色,以监测精液衍生淀粉样蛋白纤维的形成。通过圆二色光谱测量PAP248 - 286的β折叠结构形成,并使用透射电子显微镜(TEM)观察有无乳酸孵育的淀粉样蛋白纤维的形态。使用病毒感染试验确定不同时间点与乳酸孵育的淀粉样蛋白纤维的增强作用。将PAP248 - 286(2 mg/mL)与健康女性阴道分泌物稀释液孵育,并用ThT和刚果红染色检测淀粉样蛋白纤维的形成。
乳酸在体外以剂量依赖方式抑制SEVI纤维形成。0.5 mg/mL的乳酸在48小时内完全抑制2 mg/mL SEVI纤维形成。孵育48小时后,1 mg/mL的乳酸抑制了SEVI(2 mg/mL)β折叠结构的形成,并如TEM观察到的那样完全抑制了2 mg/mL PAP248 - 286的聚集。在乳酸存在下,PAP248 - 286失去了增强病毒感染的能力。阴道分泌物以剂量依赖方式抑制SEVI纤维形成,并且在将2 mg/mL PAP248 - 286与67%阴道分泌物孵育后几乎没有SEVI纤维形成。
乳酸在体外抑制SEVI纤维形成。