木瓜蛋白酶对凝血功能的抑制作用及机制的体外研究

目的：观察木瓜蛋白酶体外对凝血功能的抑制作用,并探讨其可能机制。方法：将不同剂量木瓜蛋白酶分别与贫血小板血浆（PPP）和富血小板血浆（PRP）作用,分为生理盐水组、10U/L组和20U/L组,分别以血凝仪测定PPP和PRP的凝血酶原时间（PT）和活化部分凝血活酶时间（APTT）,以血气分析仪和血凝仪分别测定PPP的Ca2＋浓度和凝血因子V、VII、VIII、IX、X和XI活性（FV：C、FVII：C、FVIII：C、FIX：C、FX：C和FXI：C）;将新鲜全血与前述3种浓度木瓜蛋白酶作用,采用硅化管法测定全血凝血时间（CT）。同时测定0、20、40、60和80U/L木瓜蛋白酶PPP的PT和APTT值。结果：10 U/L和20U/L木瓜蛋白酶组PPP和PRP的PT和APTT值、全血CT值分别显著高于生理盐水组和10U/L木瓜酶组（P〈0.01）,FV：C和FVIII：C水平分别显著低于生理盐水组和10U/L木瓜酶组（P〈0.05）;三组PPP与PRP之间PT和APTT值、各组间Ca2＋浓度以及其余凝血因子活性差异均无显统计学意义（P〉0.05）。PPP的PT和APTT值均与木瓜蛋白酶剂量呈显著正相关（r=0.995和0.991,P〈0.01）。结论：木瓜蛋白酶可通过抑制凝血因子V和VIII活性,从而对凝血功能有剂量依赖性的抑制作用,具有抗凝的功效。     

An ultrastructural study on the ear cartilage of rabbits after the administration of papain

Summary Crude papain was administered intravenously to young rabbits and the cartilage of the collapsed ear was examined electron-microscopically. Degeneration and recovery of chondrocytes, and decrease in and recovery of the electron-density of elastic fibers, were observed during the collapse and restoration of the ear. Some samples were stained with ruthenium red. In the collapsed ear, with a marked decrease of proteoglycan in the cartilage, loss of ruthenium red-positive granules was observed in the extracellular matrix. Collagen fibrils in the cartilage appeared to be somewhat increased in number, some of their diameters became slightly greater, and a part were assembled into bundles, occasionally accompanied by periodic crossstriation. Decrease of proteoglycan in the cartilage matrix probably brought about the unmasking and the assembly of collagen fibrils. In one of the experimental animals, collagen fibrous segments of an atypical fibrous long spacing (FLS-)type with symmetrical cross-striation were found around the chondrocytes in the ear cartilage, during the period of recovery. Some kind of the endogenous sulfated carbohydrate may have acted to affect the arrangement of type II collagen or procollagen molecules newly produced by the recovering chondrocytes.     

Low-molecular-weight proteinase inhibitor of human epidermis inhibiting papain and trypsin activity

Summary A low-molecular-weight proteinase inhibitor was isolated from human epidermal extracts by means of ultrafiltration, gel filtration and ion-exchange chromatography. On polyacrylamide gel electrophoresis (PAGE), the isolated fraction exhibited a single band of activity at pH 9.4, and inhibitory activity against papain and trypsin was detected in this band. Using sodium-dodecyl-sulphate PAGE, the molecular mass of the inhibitor was estimated to be 2,200 daltons.     

Comparison of two different benzodiazepine binding proteins by peptide mapping after limited proteolysis

Proteins P₅₁ and P₅₅ were photolabeled by [³H]flunitrazepam, [³H]clonazepam or ³H-Ro 15–4513 and then compared by peptide mapping after limited digestion with various proteases. Results indicated that [³H]flunitrazepam or [³H]clonazepam irreversibly bind to the same and [³H]Ro 15–4513 to a different part of these proteins. The different radiolabeled peptide patterns obtained from P₅₁ or P₅₅ irrespective of the photolabel used, suggest a difference in the molecular structure of these proteins.     

云南峨山番木瓜中木瓜蛋白酶的初步纯化及活性测定

应用FPLC系统对采自云南峨山县的番木瓜乳汁中的木瓜蛋白酶进行了初步纯化,同时,用基梅尔-史密斯法测定了番木瓜乳汁中木瓜蛋白酶的活性。结果表明:不同瓜形之间的木瓜蛋白酶活性大小没有明显差异,但与瓜的成熟度有关。     

IL‐1R1–MyD88 axis elicits papain‐induced lung inflammation

Allergic asthma is characterized by a strong Th2 response with inflammatory cell recruitment and structural changes in the lung. Papain is a protease allergen disrupting the airway epithelium triggering a rapid inflammation with eosinophilia mediated by innate lymphoid cell activation (ILC2) and leading to a Th2 immune response. In this study, we focused on inflammatory responses to a single exposure to papain and showed that intranasal administration of papain results in the recruitment of inflammatory cells, including neutrophils and eosinophils with a rapid production of IL‐1α, IL‐1β, and IL‐33. The inflammatory response is abrogated in the absence of IL‐1R1 and MyD88. To decipher the cell type(s) involved in MyD88‐dependent IL‐1R1/MyD88 signaling, we used new cell‐specific MyD88‐deficient mice and found that the deletion of MyD88 signaling in single cell types such as T cells, epithelial cells, CD11c‐positive or myeloid cells leads to only a partial inhibition compared to complete absence of MyD88, suggesting that several cell types contribute to the response. Importantly, the inflammatory response is largely ST2 and IL‐36R independent. In conclusion, IL‐1R1 signaling via MyD88 is critical for the first step of inflammatory response to papain.     

The effect of proteolytic enzymes on hair follicles of transgenic mice expressing the lac Z-protein in cells of the bulge region

OBJECTIVE: To study the effects of proteolytic enzymes on mice hair follicles, particularly on cells of the bulge area regarded as follicle stem cells. BACKGROUND: Previous application by iontophoresis of proteolytic enzymes on guinea pig skin resulted in degenerative effects on hair follicles and the hypothesis was proposed that some of the affected cells could be stem cells. METHODS: To mark putative stem cells transgenic mice were produced carrying the lac-Z gene fused to the Upstream Regulatory Region (URR) of Human Papilloma Virus 11 (HPV11), as they express this gene specifically in the cells of the bulge area. Chymotrypsin and papain were applied on skin by iontophoresis, trypsin in the form of liposomes. RESULTS: Enzyme application, both by electrophoresis and as liposomes, led to intense degenerative effects of the hair follicle, such as detachment of the inner root sheath, cystic dilation of the hair shaft and presence of epithelial cells within the lumen. Some of these cells represent hair follicle stem cells expressing beta-galactosidase (beta-gal), having been detached from the bulge area as a result of enzyme treatment, implying impairment of their function.     

兔肺气肿模型的建立

目的：应用木瓜蛋白酶雾化吸入和静脉注射法来复制兔肺气肿模型。方法：兔耳缘静脉注入8％木瓜蛋白酶和让兔雾化吸入5％木瓜蛋白酶液制作模型。观测其动脉血气及肺功能并对肺组织进行病理学观察。结果：与正常组相比模型组兔动脉血pH值、PaO2浓度降低，PaCO2浓度升高；潮气量下降，呼吸总阻力、中心阻力、总气道阻力升高；病理组织切片光镜下见肺泡壁变薄、断裂，肺泡腔扩张、融合，典型肺气肿表现。结论：木瓜蛋白酶雾化吸入和静脉注射能成功复制出兔肺气肿模型。     

Effectiveness of papain gel in venous ulcer treatment: randomized clinical trial

OBJECTIVE:

to assess the effectiveness of 2% papain gel compared to 2% carboxymethyl cellulose in the treatment of chronic venous ulcer patients.

METHOD:

randomized controlled clinical trial with 12-week follow-up. The sample consisted of 18 volunteers and 28 venous ulcers. In the trial group, 2% papain gel was used and, in the control group, 2% carboxymethyl cellulose gel.

RESULTS:

the trial group showed a significant reduction in the lesion area, especially between the fifth and twelfth week of treatment, with two healed ulcers and a considerable increase in the amount of epithelial tissue in the wound bed.

CONCLUSION:

2% papain gel demonstrated greater effectiveness in the reduction of the lesion area, but was similar to 2% carboxymethyl cellulose gel regarding the reduction in the amount of exudate and devitalized tissue. Multicenter research is suggested to evidence the effectiveness of 2% papain gel in the healing of venous ulcers. UTN number: U1111-1157-2998