Pharmacological effects of bioactive agents in earthworm extract:A comprehensive review

This review compiles information from the literature on the chemical composition,pharmacological effects,and molecular mechanisms of earthworm extract(EE)and suggests possibilities for clinical translation of EE.We also consider future trends and concerns in this domain.We summarize the bioactive components of EE,including G-90,lysenin,lumbrokinase,antimicrobial peptides,earthworm serine protease(ESP),and polyphenols,and detail the antitumor,antithrombotic,antiviral,antibacterial,anti-i nflammatory,analgesic,antioxidant,wound-healing,antifibrotic,and hypoglycemic activities and mechanisms of action of EE based on existing in vitro and in vivo studies.We further propose the potential of EE for clinical translation in anticancer and lipid-modifying therapies,and its promise as source of a novel agent for wound healing and resistance to antibiotic tolerance.The earthworm enzyme lumbrokinase embodies highly effective anticoagulant and thrombolytic properties and has the advantage of not causing bleeding phenomena due to hyperfibrinolysis.Its antifibrotic properties can reduce the excessive accumulation of extracellular matrix.The glycolipoprotein extract G-90 can effectively scavenge reactive oxygen groups and protect cellular tissues from oxidative damage.Earthworms have evolved a well-developed defense mechanism to fight against microbial infections,and the bioactive agents in EE have shown good antibacterial,fungal,and viral properties in in vitro and in vivo experiments and can alleviate inflammatory responses caused by infections,effectively reducing pain.Recent studies have also highlighted the role of EE in lowering blood glucose.EE shows high medicinal value and is expected to be a source of many bioactive compounds.

Hypothetical mode of action of earthworm extract with hepatoprotective and antioxidant properties

The hepatoprotective potential of earthworm extract (EE) (Lampito mauritii, Kinberg) was evaluated against paracetamol-induced liver injury in Wistar albino rat, in comparison with silymarin, the standard hepatoprotective drug. We observed a reduction in liver antioxidants, such as glutathione (GSH), superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) and in serum total protein, and an increase in serum alkaline phosphatase (ALP), serum aspertate aminotranferase (AST), serum alanine aminotranferase (ALT), bilirubin and liver thiobarbituric acid reactive substances (TBARS) due to liver injury in the paracetamol-administered rats (2 g/kg). On the contrary, increased activities of liver GSH, SOD, GPx, CAT and serum total protein level, and decrease in the contents of serum ALP, AST, ALT, bilirubin and liver TBARS were observed in rats administered with different doses of EE (100, 200 and 300 mg/kg), which are similar to the activities of hepato-protective drug silymarin (150 mg/kg). The mode of action of EE as evidenced by the above parameters may suggest that EE, on the one hand, prevents the formation of the reactive oxygen groups, or scavenges these groups, thereby preventing the damage on the hepatic cells, and, on the other hand, modulates the genes responsible for synthesis of antioxidant enzymes such as GPx, CAT and SOD in liver tissue and decreases the serum enzymatic activities such as ALP, AST and ALT.

赤子爱胜蚓油脂的提取及脂肪酸组成分析

为促进蚯蚓蛋白粉加工副产物的开发与利用,以赤子爱胜蚓渣料为原料,采用超声辅助水酶法结合酸水解法提取蚯蚓油,研究酶种类、酶解时间和加酶量对蚯蚓油得率的影响,通过正交实验确定最佳酶解条件。同时,对提取的蚯蚓油理化指标及脂肪酸组成进行分析。结果表明:超声辅助水酶法结合酸水解法提取蚯蚓油的最佳酶解条件为使用胃蛋白酶(适宜温度50℃、pH 2.0)、酶解时间2.0 h、加酶量4%,在此条件下蚯蚓油得率为11.45%;蚯蚓原油为浅棕色,稍有浑浊,稍有蚯蚓特有的土腥味,略带有油脂酸败味;蚯蚓油由24种脂肪酸构成,其中饱和脂肪酸含量较高,占总脂肪酸的82.381%,主要为硬脂酸、月桂酸,单不饱和脂肪酸占总脂肪酸的8.565%,主要为油酸、棕榈油酸,多不饱和脂肪酸占总脂肪酸的8.995%,主要为亚油酸,另外蚯蚓油奇数碳脂肪酸含量为13.867%。综上,超声辅助水酶法结合酸水解法对于提取蚯蚓渣料中的油脂有一定效果,具备实际应用的潜力,且得到的蚯蚓油含有丰富的奇数碳脂肪酸,营养价值较高。

基于土壤PAHs形态的老化效应与生物有效性研究

土壤多环芳烃(PAHs)不同提取形态可为理解老化效应机理和评估生物有效性提供新的视角,文章选用芜湖市4种不同类型农业土壤,通过连续提取方法将土壤PAHs划分为3种可提取形态,在此基础上探究PAHs在土壤中的老化效应与生物有效性。结果表明,在4种农业土壤中PAHs含量均表现出慢解吸态>快解吸态>水溶态的特征。在外源PAHs进入土壤的第0天,慢解吸态占比就已达到55.61%±21.59%,有机质含量最低的土壤中,第0天慢解吸态比例明显小于其他土壤,说明这部分“快吸附、慢解吸”的PAHs是与有机质密切联系的。随着外源PAHs在土壤中老化时间的延长,慢解吸态持续增加至第90天的75.57%±7.43%,而水溶态和快解吸态分别从11.20%±4.25%和33.19%±20.24%下降至0.89%±0.21%和1.51%±1.11%,质量平衡计算得出的不可提取态则达到22.0%±6.8%,表明PAHs在土壤中老化效应实质上是从水溶态和快解吸态向慢解吸态乃至不可提取态转化的过程。这种转化过程的动力学可以用一阶指数模型很好地模拟。不同提取形态PAHs中,水溶态含量与蚯蚓富集实验得出的土壤PAHs生物富集系数相关性最强(R=0.84,P<0.01),表明水溶态PAHs可以作为评价土壤PAHs生物有效性的替代指标。

Radioenhancing effect of earthworm capsule medication on radiotherapy in the treatment of patients with carcinoma of esophagus or lung

Earthworm capsule is an extract from earthworm made in our laboratory.From1986 to 1989,486 patients with carcinoma of esophagus or lung were randomly divided into 2groups,one group received radiotherapy alone(control group)and the other radiotherapycombined with earthworm capsules(treatment group).All cases were diagnosedpathologically or cytologically.In a two-month treatment course,earthworm capsuleswere given orally twice a day,2～3 capsules each time,each capsule containing 60 mg ofthe extract.Compared with the control group,the CR(complete remission)rate andCR+PR(partial remission)rate in patients with carcinoma of esophagus treated byradiotherapy in combination with earthworm capsule increased 9.7%(P【0.01) and 9.6%(P【0.05),while in patients with carcinoma of lung,they increased 10.6%(P【0.05)and13.5%(P【0.05),respectively.The earthworm capsule caused irritation in the alimentarytract,the patients displaying slight nausea and vomiting(16.7% in thecombination treatment group and 10.0% in the control group).However,these side ef-fects could be alleviated when capsules were taken after meal or with honey concomitantly.No deleterious effects on the heart,liver,kidney,skin,hematopoietic or nervous systemwere seen.

In <i>Vitro</i>Anthelmintic Activity of Leaf Extracts of <i>Celosia</i>laxa Schum. &Thonn

Background: Celosia laxa (Amaranthaceae) is mainly a west African plant species used in Traditional Medicine as an anthelmintic, anticancer, antibacterial and analgesic agent. The seeming prevalence of worm infection in Gwandu community of Gwandu Local Government Area of Kebbi State-Nigeria prompted this study. Methods: Whole plant of C. laxa extracts obtained through maceration in aqueous (AE), methanol (ME) and hexane (HE) solvents were investigated for their anthelmintic activity against Indian earthworms (Pheretima posthuman) at four different (gradient) concentrations of 10, 20, 40 and 80 (mg/ml) for each extract. The study involved the determination of time of paralysis (P) and time of death (D) of the worms. Results: Both the aqueous and ethanolic extracts exhibited significant anthelmintic activity at the highest concentration of 80 mg/ml as compared to the standard drug, praziquantel (10 mg/ml). Consequently, the aqueous extract showed a higher activity at 80 mg/ml compared to standard praziquantel at 10 mg/ml (with no significant value of p < 0.05). The time of paralysis and death observed for AE was 13.0 ± 1.8 and 16.8 ± 1.5 while the ME was less bioactive with 15.7 ± 0.5 and 23.0 ± 0.0 respectively. However, on the other hand, the hexane extract recorded no-activity on all the test sample concentrations, compared to the standard drug (with a significant difference of p-value, p > 0.05). Conclusion: It was concluded that the leaves of C. laxa are likely to yield a potent anthelminthic drug owing to soluble phytoconstituent which are largely hydrophilicity extracted by the polar solvents. Also, considering that the plants’ mode of preparation for use by the locals was aqueous decoction before administration, the folkloric therapeutic claims can be said to have been justified.

蚯蚓提取物对葡聚糖硫酸钠诱导小鼠溃疡性结肠炎的影响

【目的】探究蚯蚓提取物对葡聚糖硫酸钠(dextran sulfate sodium,DSS)诱导的小鼠溃疡性结肠炎(ulcerative colitis,UC)的影响,为蚯蚓用于UC潜在治疗药物提供试验依据。【方法】C57BL/6小鼠自由饮用3.5%DSS溶液7 d并灌胃不同浓度的蚯蚓提取物,每日监测小鼠精神状态、体质量和大便性状;于第10天解剖,取脾脏和结肠,制作结肠组织石蜡切片并进行苏木精—伊红染色;以荧光定量PCR检测结肠组织中肿瘤坏死因子α (TNF-α)、白细胞介素6 (IL-6)、白细胞介素1β (IL-1β)和白细胞介素10 (IL-10)的m RNA表达量;以ELISA检测结肠组织中闭锁小带蛋白1 (ZO-1)、黏蛋白2 (Muc2)和咬合蛋白(occludin)的表达量。【结果】蚯蚓提取物组能减轻小鼠体质量下降程度,降低小鼠疾病活动指数评分,减少炎性细胞浸润程度,下调IL-6、IL-1β和TNF-α的mRNA表达,上调IL-10的mRNA表达以及ZO-1、occludin和Muc2的蛋白表达量。【结论】蚯蚓提取物对DSS诱导的小鼠UC具有缓解作用,小鼠结肠组织炎性损伤和UC症状减轻。蚯蚓提取物可保护结肠黏膜的完整性和改善肠黏膜屏障功能。

蚯蚓提取物中蛋白质含量测定方法的研究

目的寻找能够精确反映蚯蚓提取物中蛋白质含量的测定方法。方法采用改良Lowry法和考马斯亮蓝染料结合法(Bradford法)测定蚯蚓提取物蛋白质含量;分别使用2法测定标准牛血清白蛋白(BSA)水溶液在碱性蛋白酶水解前后的蛋白质含量。结果改良Lowry法的测定结果明显高于Bradford法。BSA水溶液酶解前,2法测定的蛋白质含量相当;使用改良Lowry法,BSA水溶液酶解前后的蛋白测定值相近;而使用Bradford法测定BSA水溶液蛋白质含量时,酶解后的测定值较酶解前出现大幅下降。结论在测定多肽含量多的供试品中蛋白质含量时,改良Lowry法测定更为精确。蚯蚓提取物中存在大量的多肽成分,所以对其进行蛋白质含量测定时,改良Lowry法能够更加精确、真实地反映蚯蚓提取物的蛋白质含量。

菲、芘在单一和复合污染条件下不同老化阶段的提取率及其生物有效性

研究了土壤中多环芳烃菲、芘在单一和复合污染条件下的老化过程,150天内观察到菲、芘的提取率随时间延长而逐渐减少,下降速率呈现初始较快、而后减慢的趋势;菲、芘的环糊精提取法的提取率随老化时间延长而下降的趋势要比索氏提取法明显。老化过程中复合污染土壤中芘的索氏法提取率略低于单一污染土壤中芘的提取率。在老化的后期,复合污染土壤中菲的环糊精法提取率略高于单一污染土壤中菲的提取率。利用蚯蚓作为测试生物,研究两种污染物的老化过程对其生物有效性的影响,发现表征其生物有效性的蚯蚓吸收率、生物富集系数(BCF)均随老化时间的延长而下降,此外,老化过程中复合污染条件下菲的生物有效性总是大于菲作为单一污染源时的生物有效性。对污染物的环糊精法提取率和蚯蚓吸收率作相关分析发现,单一污染土壤中菲和芘的环糊精提取法的提取率与蚯蚓吸收率均呈显著正相关。

蚯蚓提取物对小鼠肿瘤动物模型的研究

目的 :研究蚯蚓提取物 (EFE)的免疫活性及抗肿瘤作用。方法 :采用小鼠移植性肿瘤S1 80 肉瘤及Heps肝癌的动物模型观察其肿瘤抑制作用。结果 :EFE对S1 80 肉瘤和Heps肝癌细胞的抑制率分别为 36 97%和 4 8 55% ;结论 :它对小鼠实体瘤细胞有明显的抑制作用 (P <0 0 1 )并提示对小鼠的细胞和体液免疫功能有显著增强作用。

蚯蚓提取液对彭县黄鸡生产性能、血清免疫指标和抗氧化指标的影响

为比较添加不同浓度蚯蚓提取液对彭县黄鸡生产性能、血清免疫指标和抗氧化指标的影响,选取健康且体重相近的100日龄彭县黄鸡(母鸡)288只,随机分成4个组,每组4个重复,每个重复18只。4个组均饲喂基础日粮,蚯蚓提取液饮水添加的稀释倍数分别为0、300、600、900倍。预饲期5 d,正试期47 d。每周记录采食量,记录生产性能。分别在105日龄和152日龄时全群称重,152日龄时检测血清免疫性能和抗氧化性能指标。结果显示:饮水添加300倍蚯蚓提取液时,能提高彭县黄鸡活重、105~152日龄阶段重、日增重、日采食量(P<0.05),血清C3、IgA、IgG、IgM和IL-6水平显著升高(P<0.05);添加300倍和600倍时,料重比显著降低(P<0.05);添加600倍时,C3、IgA和IgG水平显著升高(P<0.05),T-AOC水平显著降低(P<0.05);添加900倍时,IgA和IgG水平显著升高(P<0.05)。研究表明,本试验条件下饮水添加蚯蚓提取液的适宜稀释倍数为300倍。

蚯蚓提取物体内和体外抗肿瘤作用的研究

目的探讨蚯蚓提取物对Eca-109、Hella、K562细胞的生长抑制作用及机制。方法应用四甲基偶氮唑盐(MTT)比色分析,流式细胞仪测定细胞凋亡、细胞周期及观察小鼠体内肿瘤体积和重量。结果蚯蚓提取物Ⅰ,Ⅱ,Ⅲ中只有成分Ⅱ的高、中剂量(900,450 mg/L)对Eca-109、Hella、K562细胞有明显的抑制作用。体内300,600 mg/L灌胃小鼠对肿瘤有明显的抑制作用。Hella细胞检测有凋亡细胞出现,细胞周期检测,细胞被阻滞在G0-G1期,DNA合成受阻故使肿瘤细胞受抑制,肿瘤体积缩小。结论蚯蚓提取物Ⅱ对肿瘤细胞增殖有明显的抑制作用,主要通过细胞凋亡和肿瘤细胞受阻于G0-G1期,使DNA合成减少。

蚯蚓浸膏对环磷酰胺诱发的蚕豆根尖细胞变异的保护作用

以蚕豆根尖为材料，研究了赤子爱胜蚓浸膏对环磷酰胺诱变的保护作用．结果表明，用赤子爱胜蚓浸膏处理后，可以降低由环磷酰胺诱发的蚕豆根尖细胞微核率和染色体畸变率．１％蚯蚓浸膏处理组与阳性对照组相比，微核率和染色体畸变率分别降低７０２１％和４５３６％；０５％蚯蚓浸膏处理组，微核率和染色体畸变率分别降低４６３９％和３８３９％；０１％蚯蚓浸膏处理组，微核率和染色体畸变率分别降低２９８８％和２６８６％．

改良Lowry法和Bradford法测定蚯蚓提取物中蛋白质含量的比较(英文)

目的寻找精确反映蚯蚓提取物中蛋白质含量的测定方法。方法采用改良Lowry法和考马斯亮蓝染料结合法(Bradford法)测定蚯蚓提取物蛋白质含量;再使用两法测定标准牛血清白蛋白(BSA)水溶液在碱性蛋白酶水解前与后的蛋白质含量。结果在测定蚯蚓提取物中蛋白质含量时,改良Lowry法的测定结果明显高于Bradford法。未进行酶解的BSA水溶液,两法测定的蛋白质含量相当;使用改良Lowry法,BSA水溶液酶解前与后的蛋白测定值相近;而使用Bradford法时,测定BSA水溶液酶解后蛋白质含量的测定值较酶解前出现大幅下降。结论测定多肽含量多的生物样品中蛋白质含量时,改良Lowry法更为精确。由于蚯蚓提取物中大量多肽成分的存在,故对测定其蛋白质含量时,改良Lowry法能够更加精确、真实地反映蚯蚓提取物的蛋白质含量。

蚯蚓提取物对微循环障碍形成的保护作用

目的研究蚯蚓提取物对金黄地鼠血液微循环障碍形成的保护作用及其机理。方法以高分子右旋糖酐所致血液微循环障碍模型为基础 ,采用直观图像图示法和体外红细胞聚集试验 ,研究蚯蚓提取物对金黄地鼠颊囊血液微循环障碍的改善及体外红细胞聚集情况。结果以 2 0、1 0 0、2 0 0mg kg剂量经口给予蚯蚓提取物 3d ,可明显减少体内微血管中血细胞聚集团块和加快微血流速度 (P <0 .0 1 ) ,各观察时间点的红细胞聚集均显著低于阴性对照组 (P <0 .0 1 ) ;体外试验表明 ,在中、低切变率时 ,体外红细胞聚集状态得到明显改善 ,红细胞主要呈串珠及缗钱状 ,与阴性对照组相比有显著性统计学差异 (P <0 .0 5 )。结论蚯蚓提取物对微循环障碍形成具有一定的保护作用 。

蚯蚓提取物治疗肾实质性高血压的研究

目的 :探讨蚯蚓提取物治疗肾实质性高血压的效果及其疗效的机理。方法 :对合并高血压 18例原发性肾小球疾病和 16例终末期肾衰患者给予口服蚯蚓提取物 15 0mg/次 ,每日 3次 ,连服 14d ,每天测定血压的变化 ,并在治疗前后取血测定血浆ET - 1的含量。结果 :肾小球疾病患者血浆ET - 1含量与平均动脉压成正比 (P <0 0 5 ) ,口服蚯蚓提取物治疗后 ,平均动脉压较治疗前明显降低 (P <0 0 1) ,同时血浆ET - 1的含量测定值也明显降低(P <0 0 0 1) ,结论 :口服蚯蚓提取物具有降血压作用 。

蚯蚓提取液纤溶及抗凝作用的实验研究

通过纤维蛋白平板法和纤溶酶原激活剂活力测定试剂盒测定，证实蚯蚓提取液具有较强的纤溶活性和较高的纤溶酶原激活剂活力。动物实验结果显示，给SD大鼠静往蚯蚓提取液，能迅速提高大鼠血浆总纤溶酶原激活剂活力；口服给药也能明显减轻13％高分子葡聚糖溶液诱导大鼠实验性弥漫性血管内凝血的严重程度。

地龙2号对肝纤维化大鼠uPA和PAI-1蛋白表达的影响

目的研究蚯蚓提取物地龙2号对实验性大鼠肝纤维化的uPA和PAI-1蛋白表达的影响。方法52只雄性Wistar大鼠,随机分成6组:正常大鼠组(N);秋水仙碱对照组(CC);造模同时用秋水仙碱0.1mg.Kg-1.d-1灌胃,假模型组(NC):皮下注射花生油(CCl4),灌单蒸水;模型组对照组(MC):皮下注射CCl4花生油混合物,每日单蒸水灌胃;地龙大剂量组(Eh):造模同时用地龙2号50mg.Kg-1.d-1灌胃;地龙小剂量组(El):造模同时用地龙2号25mg.Kg-1.d-1灌胃。8周末处死大鼠,用免疫组化法检测各组大鼠肝脏组织中uPA和PAI-1的表达情况。结果地龙2号大、小剂量组与模型组相比,uPA和PAI-1的表达均显著降低,且uPA与PAI-1的表达下降呈明显的正相关。结论地龙2号可能通过抑制肝纤维化大鼠肝组织中肝星状细胞(HSC)的活化及抑制uPA、PAI-1的表达发挥抗肝纤维化作用。

蚯蚓提取物对肝纤维化大鼠α-SMA、TGFβ1、uPA及PAI-1蛋白表达的影响

目的研究蚯蚓提取物地龙2号对实验性大鼠肝纤维化的α肌动蛋白(αSMA)、转化生长因子β1(TGFβ1)、尿激酶型纤溶酶原激活物(uPA)及Ⅰ型纤溶酶原激活抑制物(PAI1)蛋白表达的影响。方法52只雄性Wistar大鼠,随机分成6组:正常组(N);阴性对照组(NC),皮下注射花生油,灌单蒸水;模型组(M),皮下注射CCl4,每日单蒸水灌胃;阳性对照组(PC),造模同时用秋水仙碱0.1mg·kg-1·d-1灌胃;地龙大剂量组(Eh),造模同时用地龙2号50mg·kg-1·d-1灌胃;地龙小剂量组(El),造模同时用地龙2号25mg·kg-1·d-1灌胃。8周末处死大鼠,用免疫组化法检测各组大鼠肝脏组织中αSMA、TGFβ1、uPA及PAI1的表达情况。结果地龙2号大、小剂量组与模型组相比,αSMA、TGFβ1、uPA及PAI1的表达均显著降低,且uPA、PAI1与αSMA及TGFβ1表达下降呈明显的正相关。结论地龙2号可能通过抑制肝纤维化大鼠肝组织中肝星状细胞的活化,降低TGFβ1、uPA及PAI1的表达,发挥抗肝纤维化作用。

蚯蚓提取物对小鼠脾脏抗体形成细胞的影响

作者用定量溶血分光光度法(QHS)同时测定正常和荷瘤BALB/c小鼠各40只脾脏抗体形成细胞的变化,观察蚯蚓提取物对B细胞介导的免疫应答的影响。结果表明,蚯蚓提取物连续灌胃7d后,使正常小鼠脾脏抗体形成细胞由1.117±0.035增至1.291±0.103(OD值)(P<0.01);荷瘤小鼠由 l.045±0.062增至1.343±0.173(P<0.01)。提示蚯蚓提取物能调节B细胞的增殖和分化,使特异性抗体形成和分泌增加。