Guangdong Provincial Key Laboratory of Marine Biotechnology, STU-UNIVPM Joint Algal Research Center, Department of Biology, College of Science, Shantou University, Shantou, 515063, Guangdong, PR China.
Biotechnology Research Centre, MARDI, P.O.Box 12301, 50744, Kuala Lumpur, Malaysia.
J Ethnopharmacol. 2021 Jun 28;274:114024. doi: 10.1016/j.jep.2021.114024. Epub 2021 Mar 13.
Lignosus rhinocerotis (Cooke) Ryvarden cultivar TM02, also known as tiger's milk mushroom, is regarded as important folk medicine in Malaysia, while is used for the treatment of liver cancer, chronic hepatitis, gastric ulcer in traditional Chinese medicine. However, there is no compilation of scientific evidence that its protection for gastric, and no attempts have been made to understand how polysaccharides in Lignosus rhinocerotis might promote intestinal mucosal wound healing.
This study aimed to investigate the effect and mechanism of β-glucan prepared from L. rhinocerotis using an enzymatic method on epithelial restitution during intestinal mucosal damage.
Based on FT-IR, MALDI-TOF-MS, HPSEC-MALLS-RID, and AFM, the structure of polysaccharides from L. rhinocerotis was analysed. In addition, polysaccharides were used to test for wound healing activity in IEC-6 cells by measuring cell migration, proliferation, and expression of cell division control protein 42, Rac-1, RhoA, and Par-3.
β-glucan was extracted using enzyme-assisted extraction, and a yield of approximately 8.5 ± 0.8% was obtained from the dried biomass. The β-glucan extracted by enzyme-assisted extraction (EAE) of polysaccharides was composed entirely of D-glucose with a total carbohydrate content of 95.5 ± 3.2%. The results of HPLC, FTIR, and MALDI-TOF-MS analyses revealed EAEP to be confirmed as β-glucan. The molecular weight of prepared β-glucan was found to be 5.315 × 10 g/mol by HPSEC-MALLS-RID. Furthermore, mucosal wound healing studies showed that the treatment of IEC-6 with a β-glucan concentration of 200 μg/mL promoted cell migration and proliferation, and it enhanced the protein expression of cell division control protein 42, Rac-1, RhoA, and Par-3.
The present study reveals that the prepared β-glucan accelerates intestinal epithelial cell proliferation and migration via activation of Rho-dependent pathway. Hence, β-glucan can be employed as a prospective therapeutic agent for the treatment of diseases associated with gastrointestinal mucosal damage, such as peptic ulcers and inflammatory bowel disease.
裂蹄木层孔菌（Cooke）Ryvarden 栽培品种 TM02，也被称为虎奶菇，在马来西亚被视为重要的民间药物，而在中国传统医学中则用于治疗肝癌、慢性肝炎、胃溃疡。然而，目前尚无科学证据表明其对胃有保护作用，也没有人试图了解裂蹄木层孔菌中的多糖如何促进肠道黏膜伤口愈合。
本研究旨在探讨酶法制备裂蹄木层孔菌β-葡聚糖对肠道黏膜损伤时上皮修复的作用及机制。
基于傅里叶变换红外光谱（FT-IR）、基质辅助激光解吸电离飞行时间质谱（MALDI-TOF-MS）、高效凝胶渗透色谱-多角度激光散射-折射率（HPSEC-MALLS-RID）和原子力显微镜（AFM），对裂蹄木层孔菌多糖的结构进行了分析。此外，还通过测定细胞迁移、增殖以及细胞分裂控制蛋白 42、Rac-1、RhoA 和 Par-3 的表达，检测了 IEC-6 细胞中多糖的伤口愈合活性。
采用酶辅助提取法提取β-葡聚糖，从干燥生物量中可获得约 8.5±0.8%的收率。酶辅助提取的裂蹄木层孔菌多糖（EAE）β-葡聚糖完全由 D-葡萄糖组成，总糖含量为 95.5±3.2%。HPLC、FTIR 和 MALDI-TOF-MS 分析结果证实，EAEP 被确认为β-葡聚糖。HPSEC-MALLS-RID 分析表明，制备的β-葡聚糖的分子量为 5.315×10^5g/mol。此外，黏膜伤口愈合研究表明，浓度为 200μg/mL 的β-葡聚糖处理 IEC-6 可促进细胞迁移和增殖，并增强细胞分裂控制蛋白 42、Rac-1、RhoA 和 Par-3 的蛋白表达。
本研究揭示了制备的β-葡聚糖通过激活 Rho 依赖性通路加速肠道上皮细胞增殖和迁移。因此，β-葡聚糖可用作治疗与胃肠道黏膜损伤相关疾病的潜在治疗药物，如消化性溃疡和炎症性肠病。
