Influence of Hellebores Ingredients (Helleborus odorus Waldst. et Kit.) on Hematological Parameters, Acute Phase Proteins and Neutrophil Granulocytes Functions in Rats

Beograd : [V. Davidović]

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Veterinarska medicina - Fiziologija / Veterinary medicine - Physiology

U okviru ove doktorske disertacije je ispitivan uticaj sastojaka petroletarskog (PEK), hloroformskog (HEK), metanolnog (MEK) i vodenog ekstrakta (EK) rizoma i korena Helleborus odorus Waldst. et Kit. na hematološke parametre, odgovor akutne faze (brzinu sedimentacije i koncentraciju proteina akutne faze) i funkcije neutrofilnih granulocita (stepen fagocitoze i intenzitet oksidativnog praska) kod Wistar pacova. Kontrolnoj grupi je intramuskularno (IM) aplikovan sterilan fiziološki rastvor u količini od 2,5 ml/kg TM. Ogledne grupe su IM tretirane različitim ekstraktima kukureka u dozama od 50, 100 ili 200 mg/kg TM, a krv za analizu je uzimana posle 24h. Osim toga, ispitivan je i uticaj vodenog i metanolnog ekstrakta aplikovanih 24h nakon trokratnog tretmana pacova deksametazonom u dozi od 1 mg/kg TM, na uočene efekte. Ogled je izveden na ukupno 147 pacova podeljenih u 21 grupu od po 7 jedinki. Aplikovanje različitih doza (50, 100 ili 200 mg/kg TM) metanolnog i vodenog ekstrakta je dovelo do značajnog povećanja broja ukupnih leukocita, broja i procenta neutrofilnih granulocita, srazmernog primenjenoj dozi. Ove promene su bile izražene i kada se ekstrakti primenjuju 24h nakon aplikacije deksametazona. Aplikovanje ovih ekstrakata je imalo za posledicu smanjenje broja i procenta limfocita i povećanje neutrofilno/limfocitnog indeksa u svim oglednim grupama. Srednje vrednosti navedenih parametra su se neznatno menjale posle aplikovanja petroletarskog i hloroformskog ekstrakta. Ispitivani ekstrakti nisu uticali na broj i procenat monocita, ali je povećanje broja ovih krvnih elemenata registrovano kod primene MEK i EK nakon tretmana pacova deksametazonom. Upotrebljeni ekstrakti nisu ispoljili hemolitičku aktivnost, koncentracija hemoglobina je bila značajno povećana, a broj trombocita smanjen. Značajno bržu sedimentaciju u svim posmatranim intervalima su imali eritrociti pacova oglednih grupa kojima su aplikovani pojedinačni ekstrakti kukureka ili u kombinaciji sa deksametazonom, a najizraženiji efekat su ispoljili metanolni i vodeni ekstrakt. Statistički značajno smanjenje koncentracije albumina i povećanje koncentracija fibrinogena je utvrđeno nakon pojedinačnog aplikovanja MEK i EK ili u kombinaciji sa deksametazonom. Tretman oglednih grupa petroletarskim i hloroformskim ekstraktom nije uticao na koncentraciju albumina, a ispoljio je negativan efekat na koncentraciju fibrinogena. Aplikovanje MEK i EK je dovelo do značajnog povećanja koncentracije haptoglobina, a sa povećanjem aplikovane doze, povećavala se i vrednost ovog parametra. Razlika u prosečnoj vrednosti koncentracije haptoglobina kod pacova kojima je trokratno aplikovan deksametazon nije bila signifikantna u odnosu na grupe koje su dodatno tretirane navedenim ekstraktima. Upotrebljeni metanolni i vodeni ekstrakt su doveli do značajnog povećanja stepena fagocitoze i intenziteta oksidativnog praska neutrofilnih granulocita. Akutna zapaljenska reakcija se razvija 24h posle pojedinačnog aplikovanja različitih doza metanolnog i vodenog ekstrakta ili u kombinaciji sa deksametazonom, ali ne i nakon tretmana petroletarskim i hloroformskim ekstraktom.

Within the framework of this PhD thesis, we investigated effects that ingredients of petroleum ether (PHE), chloroform (CHE), methanol (MHE) and aqueous extract of rhizome and root of Helleborus odorus Waldst. et Kit. (HE) might have on hematological parameter values, acute phase response (sedimentation rate and concentration of acute phase proteins) and neutrophil granulocytes functions (intensity of phagocytosis and oxidative burst) in Wistar rats. Sterile saline, in the amount of 2,5 ml/kg BW, was applied intramuscularly (IM) to the animals in a control group. The experimental groups were treated intramuscularly with different Hellebores extracts in a doses of 50, 100 or 200 mg/kg BW and blood was collected following 24 hrs. We have also investigated the influence of methanol and aqueous extracts injected 24 hrs after triple treatments of rats with dexamethasone in a dose of 1 mg/kg BW on the parameters enlisted above. The experiment was conducted on the total of 147 rats divided in 21 groups consisting of 7 animals each. Application of different MHE and HE doses (50, 100 or 200 mg/kg BW) resulted in significant increase of total leukocyte count, number and percent of neutrophil granulocytes and this efect was dose dependent. These effects were also evident 24 hrs following dexamethasone treatment. In all experimental groups, lymphocyte count and percent of lymphocytes was decreased while neutrophil/lymphocyte index was increased. Other two extracts (PHE and CHE) did not exert significant effects on the above mentioned parameters. Hellebores extracts didn't influence monocyte number and percent, but increase in their number was registered after treatment with MHE and HE following dexamethasone injections. Thrombocyte count was decreased. Applied extracts didn't exert hemolitic activity and hemoglobin concentration was significantly increased. Sedimentation rate in all observed intervals was significantly increased in all groups treated with Hellebores extracts alone or in combinations with dexamethasone. Methanol and aqueous extract exerted were the most effective. Statistically significant decrease in albumin concentration and elevation of fibrinogen concentration was registrated after single application of MHE and HE alone or in combination with dexamethasone. Treatment of experimental groups with petroleum ether and chloroform extract didn't elevate albumin concentration, and exerted negative influence on the fibrinogen concentration. Application of MHE and HE resulted in significant haptoglobin concentration increase and this effect was dose dependent. Differences in haptoglobin mean values following triple treatment with dexamethasone didn't significantly differ when compared to the values recorded in groups aditionally treated with Hellebores extracts. Intensity of phagocytosis and oxidative burst by neutrophils were significantly increased with MHE and HE. Acute inflammatory reaction developed 24 hrs following application of different MHE and HE doses alone, or in combination with dexamethasone, but not and after treatment with PHE and CHE.

akutna zapaljenska reakcija, funkcije neutrofilnih granulocita, hematološki parametri, kukurek, pacovi

acute inflammatory reaction, Helleborеs, hematological parameters, neutrophil granulocytes functions, rats

619:616:636

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