Effects of hyaluronidase (HYAL) and Spiperone treatment on collagen type I, hydroxyproline, total soluble collagen, hyaluronic acid levels after the single dose intratracheal bleomycin instillation.

Hydroxyproline, collagen type I, total soluble collagen and hyaluronic acid levels were measured in homogenate of right lung lobes from C57BL/6 mice on 21st day after BLM treatment. Hydroxyproline, collagen type I and HA were assayed by ELISA according to manufacturer instructions (Cusabio Biotech CO., Ltd, China). The right lung lobes were excised and snap frozen after having measured the wet weight. Sensitivities were >1.95 ng/mL for hydroxyproline, >0.039 ng/mL for collagen type I and >15.6 pg/mL for HA. The total soluble collagen was determined using a standard curve for the SircolTM assay (Biocolor Ltd, UK) according to manufacturer’s instructions. Results were expressed as μg collagen per right lung.
Data represent mean of 3 independent experiments, n = 5/group. Results are presented as mean and SEM.
*—compared to the mice received intratracheal 0.9% NaCl (P <0.05),
●—compared to the mice received intratracheal BLM and 0.9% NaCl treated (P<0.05), t test was used.
Effects of hyaluronidase (HYAL) and Spiperone treatment on collagen type I, hydroxyproline, total soluble collagen, hyaluronic acid levels after the single dose intratracheal bleomycin instillation.

本研究检测了博莱霉素(BLM)造模后第21天的C57BL/6小鼠右肺叶匀浆中羟脯氨酸、I型胶原、总可溶性胶原及透明质酸(hyaluronic acid, HA)的水平。采用酶联免疫吸附测定(ELISA)法，按照试剂盒说明书（中国Cusabio Biotech CO., Ltd）完成羟脯氨酸、I型胶原及HA的检测。右肺叶在称量湿重后被切除并快速冷冻。各指标的检测灵敏度分别为：羟脯氨酸＞1.95 ng/mL，I型胶原＞0.039 ng/mL，HA＞15.6 pg/mL。总可溶性胶原采用英国Biocolor Ltd的Sircol™胶原检测试剂盒，依照说明书以标准曲线法定量，结果以每只小鼠右肺的胶原微克数表示。
数据来源于3次独立重复实验，每组n=5。结果以平均值±标准误(standard error of the mean, SEM)呈现。
*— 与气管内注射0.9%氯化钠溶液的小鼠相比（P<0.05）；
●— 与气管内注射博莱霉素后给予0.9%氯化钠溶液处理的小鼠相比（P<0.05），统计分析采用t检验。
气管内单次滴注博莱霉素后，透明质酸酶(HYAL)与螺哌酮处理对I型胶原、羟脯氨酸、总可溶性胶原及透明质酸水平的影响。