血小板激活因子与生殖生理

血小板激活因子(platelet activating factor,PAF)是一种独特的磷脂,具有广泛的生物学活性,在炎症、休克、哮喘、溃疡等疾病的发生中起重要作用。近年来发现,PAF还参与生殖生理过程,本文就这方面的进展作简要概述。 1 生殖器官中PAF的生理性来源 1988年Kumar首先报道丁兔精液中含有PAF,浓度可达3.5×10~(-6)mol/10~6个精子,而精浆无PAF活性。Kuzan在1989年测得刚液化的人精液PAF浓度为0.5×10~(-6)mol/10~6个精子,经37℃、95％CO_2孵育24小时后,PAF浓度可升高50余倍。Muguruma等报道了豚鼠和大鼠雄性生殖器官中PAF总活性分布,结果以附睾及睾丸最高,附睾为精子成熟部位,此处高PAF活性可能与精子成熟有关。     

2型糖尿病大鼠不同时期膀胱收缩力动态变化的实验研究

目的观察2型糖尿病(T2DM)大鼠不同时期膀胱收缩力的动态变化情况,以探讨糖尿病膀胱病变的病理生理学机制。方法制备T2DM大鼠模型,以同龄正常大鼠为对照,在成模后6、12和20周,应用M受体激动剂卡巴胆碱,在不同药物浓度水平下进行离体逼尿肌环收缩刺激实验。结果 6周时实验组的最大收缩力(1.12±0.16)g/mg,比对照组(0.99±0.06)g/mg增高;半数有效剂量浓度(EC50)实验组(1.7±0.4)×10-7mol/L比对照组(2.7±0.3)×10-7mol/L降低;12周时实验组的最大收缩力(1.20±0.12)g/mg与对照组(1.21±0.08)g/mg差异无统计学意义;EC50实验组(2.2±0.4)×10-7mol/L与对照组(2.3±0.4)×10-7mol/L差异无统计学意义;20周时实验组的最大收缩力(1.01±0.05)g/mg比对照组(1.90±0.09)g/mg降低;EC50实验组(2.7±0.4)×10-7mol/L比对照组(2.0±0.4)×10-7mol/L增高。结论不同时期的糖尿病大鼠膀胱收缩力表现不同,随着病程的进展,膀胱逼尿肌收缩力呈现先增高后降低的趋势,对卡巴胆碱的敏感性也呈现先增高后降低的变化趋势。     

RGDS的抗血小板聚集及舒血管效应

血小板聚集是正常止血的要素,并依赖于膜糖蛋白Ⅱ_b/Ⅲ_a(GPⅡ_a/Ⅲ_a)复合物与血浆粘附糖蛋白(包括纤维蛋白原 von Willebrand 因子和 fibronectin)的相互作用。研究 GPⅡ_b/Ⅲ_a 受体与纤维蛋白结合确定了在介导其自身与 GP Ⅱ_b/Ⅲ_a 接触的纤维蛋白分子中存在两种不同的氨基酸序列。其中一种序列是 Arg-Gly-Asp(RGD),该序列不仅出现在纤维蛋白 Aα链中,也出现在 fibronectin 和 von Willebrand 因子中。为了阐明其生物学效应本文合成了RGDS,并进行了与抗栓有关的生物学评价。本研究提供的数据证实 RGDS 明显抑制 PAF或 ADP 诱发的血栓形成(对 PAF 诱导的血小板聚集抑制率为67%,RGDS 浓度为9.6×10~(-7)mol/L;对 ADP 诱导的血小板聚集抑制率为87%,RGDS 浓度为9.6×10~(-7)mol/L)。该观察与 RGDS 延缓血栓形成的发现相符。虽然 RGDS 对 ADP 比对 PAF 更敏感但它对 PAF 的抑制仍十分明显这些结果均属首次报道。本研究进一步揭示了 RGDS 有舒血管作用,而且它对大鼠主动脉肌条的舒张作用不可忽视(与对照相比1×10~(-5)mol/L 的 RGDS 在 NE 处理的鼠动脉肌条上的舒张幅度为8.08±5.0%,P<0.05)。对组织内 cGMP 的累积水平(与对照相比1±10~(-5)mol/L 的RGDS 引起 cGMP 水平增高4.68±1.9pmol/mg,P<0.05)研究结果表明,提高体内 cGMP 的水平可能是 RGDS 发挥生物学作用的机制之一。RGDS 的舒血管和抗血小板聚集两种作用均可作为设计与合成新型抗栓剂的实验依据。     

大豆磷脂对脑缺血大鼠脑中磷脂含量的影响

目的研究大豆磷脂对脑缺血大鼠脑中磷脂含量的提升作用。方法Wistar大鼠喂大豆磷脂 5d后结扎左右颈总动脉 ,测定结扎后 1、6h时的血清与大脑磷脂含量 ,并与赋型组对比。结果喂大豆磷脂大鼠血中磷脂含量明显升高 ,结扎颈总动脉 6h大脑磷脂含量上升 ,总磷脂为 (89.1± 10 .9)mg g蛋白 ,磷脂酰胆碱为 (38.5± 7.9)mg g蛋白 ,磷脂酰乙醇胺为 (2 9.5± 6 .3)mg g蛋白 ,磷脂酰肌醇为 (1.75± 0 .5 4 )mg g蛋白 ,其中总磷脂和磷脂酰胆碱含量显著高于赋型组 (P <0 .0 5 )。结论大豆磷脂有抑制脑缺血大鼠大脑磷脂含量下降作用     

6-(αα-二苯基乙酰哌嗪基苯基)-4,5-二氢-5-甲基-3(2H)-哒嗪酮对兔血小板聚集及TXB_2,cAMP的影响

6-(αα-二苯基乙酰哌嗪基苯基)-4,5-二氢-5-甲基-3(2H)-哒嗪酮(简称DMDP)是我院新合成的哒嗪酮的衍生物。DMDP可以显著抑制由花生四烯酸(AA(?),ADP和血小板活化因子(PAF)诱导的免血小板聚集,其IC_(50)分别为1.12±0.1.4.19±0.5和2.97±0.1μmol/L。实验还表明DMDP在1～500 μmol/L浓度范围内呈剂量依赖性地抑制兔血小板内血栓素B_2含量,但升高兔血小板内环腺苷酸水平,这可能是其抑制血小板聚集的作用机理之一。     

6-(αα-二苯基乙酰哌嗪基苯基)-4,5-二氢-5-甲基-3(2H)-哒嗪酮对兔血小板聚集及TXB2,cAMP的影响

6-(αα-二苯基乙酰哌嗪基苯基)-4,5-二氢-5-甲基-3(2H)-哒嗪酮(简称DMDP)是我院新合成的哒嗪酮的衍生物。DMDP可以显著抑制由花生四烯酸(AA(?),ADP和血小板活化因子(PAF)诱导的免血小板聚集,其IC₅₀分别为1.12±0.1.4.19±0.5和2.97±0.1μmol/L。实验还表明DMDP在1～500 μmol/L浓度范围内呈剂量依赖性地抑制兔血小板内血栓素B₂含量,但升高兔血小板内环腺苷酸水平,这可能是其抑制血小板聚集的作用机理之一。     

血小板激活因子在肺间质性疾病中的作用

血小板激活因子(platelet-activating factor,PAF)是一种在炎症应答反应中发挥关键作用的双层磷脂分子。动物气管内滴注PAF可出现类似于间质性肺病的症状,本文目的是简要综述PAF在这类疾病中的作用。 一、PAF的一般特点 1972年,Benveniste发现IgE致敏的家兔嗜碱粒细胞能释放一种磷脂类分子,因其具有强烈刺激血小板聚集能力,故被命名为血小板激活因子(PAF)。1979年至1980年,几个研究小组证实PAF的化学结构式为1-O-烷基-2-乙酰基-Sn-甘油-3-磷酸胆碱。     

来源于天然产物的血小板活化因子拮抗剂

PAF 血小板活化因子(1,PAF)是具有很强活性的磷脂,它由多种类型的细胞(如嗜碱粒细胞、巨噬细胞、单核细胞、血小板、肥大细胞和中性粒细胞)产生或活化。其结构经测定为1-O-十六烷基/十八烷基-2-O-乙酰-sn-甘油基-3-磷酸胆碱。 PAF与多种生物活性及途径有关,是多种生理过程的重要介质之一,包括血栓形成、过敏反应、内毒素血症休克、眼疾、胃溃疡、移植物排斥、卵子植入和哮喘等。有足够的证据证明PAF在炎症中起着重要作用。PAF是诱导血管通透性增加和胸     

冠脉内应用氨甲酰胆碱对心室复极的影响

目的探讨氨甲酰胆碱对完整动物心脏心室复极的影响。方法在4只绵羊冠状动脉回旋支内分别注入1.0 mol.L-1和2.5 mol.L-1的氨甲酰胆碱,注射时间3m in。记录回旋支支配区域心外膜心电图,测量心室的激动-恢复间期(AR I)以代表心室复极时间。静脉给予一氧化氮合成抑制剂(L-精氨酸20 mg.kg-1)后,再次冠脉内给予2.5 mol.L-1的氨甲酰胆碱。结果回旋支内灌注1.0 mol.L-1和2.5 mol.L-1的氨甲酰胆碱导致其支配区域心外膜心电图心室激动-恢复间期分别延长(38±17)m s和(58±14)m s(P<0.05)。氨甲酰胆碱所致的心室激动-恢复间期延长被L-精氨酸所阻断。结论冠脉内注射氨甲酰胆碱延长心室激动-恢复间期或心室复极时间,内源性一氧化氮很可能介导氨甲酰胆碱心室复极的影响。     

枇杷叶提取物通过激活磷脂酰肌醇 3激酶 /蛋白激酶 B信号通路减轻脂多糖诱导的肺细胞损伤

目的探讨枇杷叶提取物对脂多糖( LPS)诱导的人 Ⅱ型肺泡上皮细胞 A549增殖、凋亡及氧化应激的影响及其可能作用机制。方法 2019年 6月至 2020年 7月,采用 LPS诱导的 A549细胞建立肺损伤模型( LPS组)同时将正常培养的细胞作为对照组。不同剂量( 1 mg/L、2 mg/L、4 mg/L)的枇杷叶提取物处理细胞( LPS+枇杷叶 -L组、 LPS+枇,杷叶 -M组、 LPS+枇杷叶 -H组),添加磷脂酰肌醇 3激酶( PI3K)/蛋白激酶 B(Akt)信号通路抑制剂 LY294002与枇杷叶提取物处理细胞( LPS+枇杷叶 -H+ LY294002组);采用 MTT法检测细胞增殖;采用流式细胞术检测细胞凋亡率;采用 2,4-二硝基苯肼显色法检测乳酸脱氢酶(LDH)的含量;采用黄嘌呤氧化酶法检测超氧化物歧化酶(SOD)的含量;采用硫代巴比妥酸法检测丙二醛的含量;蛋白质印迹法( Western blotting)检测磷酸化磷脂酰肌醇 -3-激酶( p-PI3K)、磷酸化蛋白激酶 B(p-Akt)、 B细胞淋巴瘤 -2(Bcl-2)、 Bcl-2相关 X(Bax)蛋白蛋白表达量。结果与对照组比较, LPS组吸光度［( 1.38±0.06)比( 0.55±0.02)］及 SOD的含量［( 81.66±5.36)U/L比(14.65±1.06)U/L］降低,凋亡率［(6.41±0.25)%比( 27.43±1.00)%］、 Bax蛋白水平及 LDH［(242.86±6.09)U/L比( 875.92±15.01)U/ L］、丙二醛［( 121.55±3.17)μmol/g比( 424.46±6.48)μmol/g］的含量升高, Bcl-2、p-PI3K［( 0.60±0.04)比( 0.13±0.01)］、 p-Akt［( 0.51±0.04)比( 0.09±0.01)］蛋白水平降低,差异有统计学意义( P<0.05);与 LPS组比较, LPS+枇杷叶 -M组、 LPS+枇杷叶 -H组吸光度［( 0.55±0.02)比( 0.86±0.03)、(1.18±0.05)］及 SOD［( 14.65±1.06)U/L比( 36.84±2.08)U/L、(70.97±3.03)U/L］的含量升高,凋亡率［( 27.43±1.00)%比( 22.24±0.81)%、(14.78±0.49)%］、 Bax蛋白水平及 LDH［( 875.92±15.01)U/L比( 641.95±9.81)U/L、(365.47±7.02)U/L］、丙二醛［( 424.46±6.48)μmol/g比( 277.94±6.90)μmol/g、(156.30±3.26)μmol/g］的含量降低, Bcl-2、p-PI3K［( 0.13±0.01)比( 0.32±0.01)、(0.54±0.03)］、 p-Akt［( 0.09±0.01)比( 0.25±0.02)、(0.42±0.03)］蛋白水平升高,均差异有统计学意义( P<0.05);添加 LY294002可明显逆转枇杷叶提取物对 LPS诱导的 A549细胞增殖、凋亡及氧化应激的作用。结论枇杷叶提取物可通过激活 PI3K/Akt信号通路减轻 LPS诱导的 A549细胞增殖抑制、凋亡和氧化应激效应,为探究枇杷叶治疗细菌感染     

Inhibition of PAF-induced eosinophil accumulation in pulmonary airways of guinea pigs by anti-asthma drugs

Intraperitoneal (i.p.) injection of platelet activating factor (PAF) in guinea pigs caused a dose-related increase in the number of eosinophils recovered from bronchoalveolar lavage fluid (BALF). The prevalence of eosinophils in BALF had significantly increased within 1 hr of i.p. injection of PAF (10 micrograms/animal) and was maximal after 24 hr. Subcutaneous osmotic mini-pumps were used to administer drugs for 5 days prior to i.p. injection of PAF (10 micrograms/animal) and for the subsequent 24 hr. The percentage increase of eosinophils in BALF, due to PAF, was inhibited in animals treated with dexamethasone, aminophylline, cromoglycate, tranilast or ketotifen, but not in animals treated with oxatomide, azelastine, amlexanox, ibudilast or AA-861. These results suggest that inhibition of pulmonary eosinophilia may be a necessary property of prophylactic anti-asthma drugs and provide indirect evidence favoring a role for PAF in eosinophilia of asthma.     

Protective effects of dazmegrel on the PAF potential of ouabain-induced cardiac arrhythmias.

The ouabain threshold to induce cardiac arrhythmias in urethane-anaesthetized guinea-pigs was not modified by the administration of either dazmegrel, 4 mg/kg i.v., or lysine-acetylsalicylate, 13.5 mg/kg i.v., 5 min before the infusion of ouabain,10g/kg per min i.v. The previous administration of platelet-activating factor (PAF), 0.01 to 1 nmol/animal i.v., 2 min prior to ouabain, caused a significant, dose-dependent decrease of the ouabain threshold for the cardiac rhythm disturbances. Lysine-acetylsalicylate lacked any effect on this PAF potentiation. Pretreatment with dazmegrel 5 min before PAF, 0.05 nmol/animal i.v., abolished the PAF potentiation of the digitalis-induced arrhythmias. These results suggest that thromboxane synthesis is involved in this PAF effect and indicate an ability of PAF to induce thromboxane generation even in the case of cyclooxygenase inhibition.     

商陆皂甙甲抑制大鼠腹腔巨噬细胞释放血小板活化因子

卡西霉素(A₂₃₁₈₇)刺激大鼠腹腔巨噬细胞释放血小板活化因子(PAF),用洗涤血小板聚集的方法测定,证明商陆皂甙甲在0.1～100μmol/L浓度范围内及所试的时间范围内,呈剂量及时间依赖性抑制大鼠腹腔巨噬细胞释放PAF:商陆皂甙甲可能就是通过抑制体内PAF生成而起抗炎作用的。     

Preclinical studies with platelet-activating factor antagonists in models of septic shock

Since the isolation and elucidation of the structure of platelet-activating factor (PAF) in the late 1970's, several preclinical studies have suggested that PAF is a key mediator of septic shock induced in animals by either endotoxin or by Gram-negative bacteria. A number of PAF antagonists have been sythesized that protect animals from the lethal effects of endotoxin. Some of these antagonists are in early stages of clinical development. The most advanced cadidate is BN 52021, a ginkgolide, that is in Phase II/III clinical trials in patients with septic shock. Preliminary results with BN 52021 indicate that it is efficacious and significantly reduces mortality associated with Gram-negative sepsis. Pivotal trials with BN 52021 aer ongoing. The present review summarizes the biological effects of PAF and the effect of PAF antagonists in animal models of septic shock. The interrelationship of PAF and tumor necrosis factor (another key mediator of septic shock) is also discussed.     

The effect of the specific PAF antagonist BN 52021 and the calcium blocker diltiazem on PAF induced arrhythmogenicity

Platelet-activating factor (PAF) in a concentration of 10(-11) mole per animal decreased the threshold doses for onsets of arrhythmia, ventricular flutters and fibrillation in ouabain induced arrhythmia in guinea-pigs in a statistically significant manner. The specific PAF antagonist BN 52021 (20 mg/kg, orally) completely inhibited the PAF effect for all types of arrhythmia, while the Ca2+ antagonistic drug diltiazem (0.1 mg/kg, i.v.) failed to counteract the PAF action. BN 52021 (20 mg/kg, per os) alone did not exert any effect on ouabain induced arrhythmia, but as expected, diltiazem (0.1 mg/kg, i.v.) showed antiarrhythmic effects. These results confirm the specific PAF antagonistic activity of BN 52021, its usefulness for PAF related cardiac rhythm disturbances and indicate that the method used could be a further useful tool to screen PAF antagonistic substances.     

Role of neutrophils in gastric damage induced by platelet activating factor

Summary Platelet-activating factor (PAF) has recently been shown to be a potent ulcerogenic agent in the stomach and intestinal mucosa. Its exact mechanism of action is not yet known although histological studies suggest that vasocongestion is an important feature of PAF-induced damage. We have therefore studied the activity of various agents with different modes of action toward PAF-induced gastrointestinal lesions in the rat (PAF 2 g/kg i.v. ; macroscopic lesions of tissues scored 20 min later; arbitrary scale from 0 to 4). Drugs were administered either i. m., s. c. (5 min) or orally (30 min) before PAF injection. PAF-induced gastric lesions were strongly inhibited by the natural PAF-antagonist BN 52021 as well as by atropine sulphate and cimetidine which implicates cholinergic stimulation in the ulcerogenic activity of PAF. The somatostatin analog BIM 23014 was also very potent against PAF, perhaps by reducing the parasympathetic stimulation in the gastric wall as described for somatostatin. Allopurinol, which is a free radical scavenger also almost totally inhibited PAF-induced gastric damage, suggesting that neutrophils are involved in the mucosal lesions. The considerable inhibition of the gastric effects of PAF found in neutrophil-depleted animal supports this hypothesis. Theophylline and disodium cromoglycate, mast cell stabilizing drugs which were also active in our model, could act by protecting mast cell degranulation induced by free radicals released from activated neutrophils. A multifunctional process seems to determine the mucosal gastric damage induced by PAF, but parasympathetic stimulation and neutrophil activation play a major role in this pathology.Send offprint requests to A. Etienne at the above address     

Cobra venom factor, an activator of the complement system, enhances the bowel necrosis induced by platelet-activating factor

We have previously produced an experimental model of ischemic bowel necrosis in the rat by injecting platelet activating factor (PAF) (7 micrograms/kg) into the mesenteric vascular bed. The dose of PAF required to produce necrosis could be reduced to 50% if the animal was pretreated with bacterial endotoxin (lipopolysaccharide) (20 micrograms/rat). The mechanism of this potentiating effect of lipopolysaccharide is unclear, but activation of the complement system may be one of the contributing factors. To investigate the role of the complement system, we injected cobra venom factor (CVF) (1 unit/kg) to activate the complement system before injection of PAF (2 micrograms/kg) into the superior mesenteric artery. CVF and PAF were also injected separately at the same dosage to other groups of rats. CVF activated the complement system, but by itself did not produce gross necrosis of the bowel. PAF alone caused 3 out of 9 rats treated to develop bowel necrosis. In contrast, combination of the two produced bowel necrosis in all 6 rats thus treated. CVF did not enhance the effects of PAF on hemoconcentration and leukopenia, but aggravated the hypotension caused by PAF. PAF, on the other hand, also enhanced activation of the complement system by CVF. To investigate the specificity of PAF on complement activation, lyso-PAF was used in combination with CVF. It was found that lyso-PAF did not have a significant potentiating effect on CVF-induced complement activation and, by itself, it had no effect on complement activation, blood pressure, white blood cell count or hematocrit. Lyso-PAF, with or without CVF, also failed to produce bowel necrosis.(ABSTRACT TRUNCATED AT 250 WORDS)     

The effect of platelet activating factor on nasal hypersensitivity

Summary Platelet activating factor (PAF) is known to have a wide range of biological activities. In the lower airways PAF has been suggested as the biochemical mediator partly responsible for the bronchial hyperreactivity which is a feature of asthma. In order to study whether PAF has a similar effect in the upper airways, we carried out a double blind, placebo-controlled cross-over study in twelve patients with strictly seasonal allergic rhinitis. The study was performed in pollen-free winter months.26 µg PAF or placebo was sprayed into each nasal cavity 8 h and 1 h before a nasal allergen challenge. The nasal response to PAF and the allergen challenge that followed was monitored by repeated measurements of nasal expiratory peak flow rate and symptom scores.PAF induced only minor changes in nasal patency and nasal symptoms as compared to placebo. However, pretreatment with PAF induced an increase in responsiveness of the nasal vasculature to the allergen challenge that followed. This was registered as a small, but statistically significant increase in the symptom scores for nasal blockage, from 1.7 (0.3; SEM) after placebo pretreatment to 2.4 (0.36; SEM) after PAF (p<0.05). A similar trend was also noted for the measurements of nasal peak flow. The other response parameters, sneezes and secretion, remained identical.These results suggest that PAF may play a role in human nasal hyperreactivity, but it appears that PAF is not a major mediator involved in the induction of this phenomenon.     

CAN THE SYNTHESIS OF PLATELET-ACTIVATING FACTOR,A POTENT VASODILATOR AND PRO-AGGREGATORY AGENT,BE ALTERED BY DIETARY MARINE OILS?

1. Diets enriched with n-3 polyunsaturated fish oils, predominantly eicosapentaenoic acid, are associated with a lower risk of atherosclerotic vascular disease. These oils purportedly reduce plasma triglycerides, total cholesterol and impair platelet aggregation. Recently, the present authors reported that rats fed a marine oil-enriched diet had significantly reduced levels of lyso-PAF, the immediate precursor of platelet-activating factor (PAF). As PAF has potent vasodilator and pro-aggregatory properties, the purpose of this study was to examine the hypothesis that fish oils affect the biosynthesis of PAF in man. 2. Supplementation of a normal diet for 3 weeks with fish oil containing the equivalent of 2.7 g of eicosapentaenoic acid daily, increased the eicosapentaenoic acid content of platelet phospholipids as well as depleting the arachidonic acid. Platelet aggregation to PAF (measured in whole blood by impedance aggregometry) was significantly impaired and whole blood thromboxane suppressed. 3. Two weeks after ceasing supplements, platelet aggregation remained impaired although thromboxane had reverted to baseline levels. There was a transient but significant fall in whole blood lyso-PAF apparent within 2 days of commencing supplements but returning to baseline levels by the end of the treatment period. Whole blood PAF followed a similar trend. 4. The effects of dietary fish oil on whole blood aggregations to PAF, on thromboxane and plasma lyso-PAF levels may be relevant to the prevention of vascular disease and the treatment of disorders in which PAF could be an inflammatory mediator.     

Anti-ulceral and anti-oxidative properties of "earthworm paste" of Lampito mauritii (Kinberg) on Rattus Norvegicus

Studies have been made to understand the anti-ulceral and anti-oxidant properties of the "earthworm paste" derived from Lampito mauritii (Kinberg), an indigenous species, in comparison with the standard anti-ulceral drug-ranitidine, on the Wistar strain albino rats Rattus norvegicus. Administration of 200 mg/kg aspirin was found to increase the volume of gastric juice secretion, total acidity, free acidity, ulcer index and reduce the pH. It also had decreased the anti-oxidant levels such as reduced glutathione, glutathione peroxidase, catalase, superoxide dismutase and increased the level of thiobarbituric acid reactive substances. Pretreatment with the standard drug-ranitidine (50 mg/kg) and different doses of "earthworm paste" (20, 40, 80, 160 and 320 mg/kg) in ulcer induced animal had enhanced the pH, decreased the volume of gastric juice, free acidity, total acidity and reduced the ulcer index. Further the activities of reduced glutathione, glutathione peroxidase, catalase, superoxide dismutase were increased whereas the thiobarbituric acid reactive substance had decreased. The results were more significant in rats administered with 160 mg/kg "earthworm paste" than the application of ranitidine and other doses of "earthworm paste". This indicates the presence of antiulcer and anti-oxidative effects in "earthworm paste". In conclusion, administration of 160 mg "earthworm paste"/kg was found to have better therapeutic properties.