Department of Studies and Research in Biochemistry, Tumkur University, Tumkur, 572103, Karnataka, India.
Department of Life Science, Jnana Bharathi Campus, Bangalore University, Bangalore, Karnataka, India.
J Ethnopharmacol. 2021 Jun 12;273:114000. doi: 10.1016/j.jep.2021.114000. Epub 2021 Mar 8.
In folk medicine, parts of Plumeria alba L. are used for the treatment of many diseases, with its latex being used for curing skin diseases and promoting wound healing.
This study aimed to study the role of P. alba L. latex in hemostasis and platelet aggregation.
The latex of P. alba L. was processed to remove waxes and enrich protein content, and the final extract was named Plumeria alba L. natant latex (PaNL). PaNL was analyzed for protease activity against casein. The type of protease in PaNL was identified by using protease inhibitors such as E-64, phenylmethylsulfonyl fluoride, ethylenediaminetetraacetic acid, and pepstatin A. Human fibrinogen, fibrin, and collagen types I and IV were subjected to hydrolysis with different concentrations of PaNL. The thrombin-like activity of PaNL was determined by analyzing its fibrinogen-clotting and procoagulant activities. The role of PaNL in platelet aggregation was also investigated. Its hemorrhagic and edema-inducing activities were evaluated in a mouse model. Phytochemical compounds were identified by gas chromatography-mass spectroscopy.
The findings of casein/gelatin zymography confirmed that PaNL possesses protease activity. The results of the protease inhibition study indicated the presence of a cysteine-type protease(s) in PaNL. PaNL hydrolyzed the subunits of fibrinogen, fibrin, and collagen types I and IV. Its fibrin-degradation activity indicated that PaNL possesses plasmin-like activity. PaNL induced clotting of citrated human plasma within 3 min of incubation in the absence of CaCl indicating the presence of thrombin-like activity, which was further confirmed by the results of the fibrinogen-clotting assay. PaNL induced platelet aggregation in the absence of agonists. There was no hemolytic activity. Mice injected with PaNL did not show edema/ hemorrhagic activity.
PaNL possesses procoagulant, fibrino(geno)lytic, thrombin- and plasmin-like activities and induces platelet aggregation, which could explain its usage for wound treatment in folk medicine.
在民间医学中，鸡蛋花的部分部位被用于治疗许多疾病，其乳胶被用于治疗皮肤病和促进伤口愈合。
本研究旨在研究鸡蛋花乳胶在止血和血小板聚集中的作用。
鸡蛋花乳胶经过处理去除蜡质并富集蛋白质含量，最终提取物命名为鸡蛋花漂浮乳胶（PaNL）。分析 PaNL 对酪蛋白的蛋白酶活性。通过使用蛋白酶抑制剂（如 E-64、苯甲基磺酰氟、乙二胺四乙酸和胃蛋白酶抑制剂 A）鉴定 PaNL 中的蛋白酶类型。用人纤维蛋白原、纤维蛋白、胶原 I 型和 IV 型进行水解，用不同浓度的 PaNL 处理。通过分析其纤维蛋白原凝固和促凝活性来确定 PaNL 的类凝血酶活性。还研究了 PaNL 在血小板聚集中的作用。在小鼠模型中评估了其致出血和致水肿活性。通过气相色谱-质谱联用鉴定了植物化学化合物。
凝块/明胶酶谱的结果证实 PaNL 具有蛋白酶活性。蛋白酶抑制研究的结果表明 PaNL 中存在一种半胱氨酸蛋白酶（s）。PaNL 水解纤维蛋白原、纤维蛋白和胶原 I 型和 IV 型的亚基。其纤维蛋白降解活性表明 PaNL 具有纤溶酶样活性。PaNL 在无 CaCl 的情况下孵育 3 分钟内诱导枸橼酸盐人血浆凝固，表明存在类凝血酶活性，纤维蛋白原凝固测定结果进一步证实了这一点。PaNL 在无激动剂的情况下诱导血小板聚集。没有溶血活性。注射 PaNL 的小鼠没有表现出水肿/出血活性。
PaNL 具有促凝、纤维蛋白（原）溶解、类凝血酶和纤溶酶样活性，并诱导血小板聚集，这可以解释其在民间医学中用于伤口治疗的用途。
