發(fā)布時(shí)間：2018-06-05 23:23

  本文選題：胰蛋白酶 + 糜蛋白酶��； 參考：《南京農(nóng)業(yè)大學(xué)》2015年碩士論文

【摘要】：胰蛋白酶、糜蛋白酶作為治療炎癥的酶類藥物,在國外臨床上已得到廣泛應(yīng)用。鑒于胰蛋白酶和糜蛋白酶在人醫(yī)臨床上的廣泛應(yīng)用,我們擬嘗試使用以胰蛋白酶和糜蛋白酶為主要組份的復(fù)合蛋白酶乳房注入劑治療奶牛乳房炎,本研究依據(jù)農(nóng)業(yè)部《獸藥研究技術(shù)指導(dǎo)原則編匯》進(jìn)行了該制劑在奶牛體內(nèi)的藥動(dòng)學(xué)、藥效學(xué)和安全性試驗(yàn)研究,為該制劑在獸醫(yī)臨床上的使用積累試驗(yàn)和理論依據(jù)。為了研究復(fù)合蛋白酶乳房注入劑在奶牛體內(nèi)藥代動(dòng)力學(xué)特征,獲取該藥的試驗(yàn)藥代參數(shù),為該藥在獸醫(yī)臨床上的使用提供劑量和方法保證,本試驗(yàn)進(jìn)行了復(fù)合蛋白酶乳房注入劑在奶牛(牛奶)的藥代動(dòng)力學(xué)研究。試驗(yàn)結(jié)果顯示,在牛奶中胰蛋白酶最低檢測限濃度為72 mu/mL,平均回收率為71.9%;糜蛋白酶最低檢測限濃度142 mu/mL,平均回收率為70.1%。同時(shí)依據(jù)本試驗(yàn)建立的檢測方法,選取受試奶牛,于每個(gè)乳區(qū)注入1支復(fù)合蛋白酶乳房注入劑,采集給藥乳區(qū)牛奶。結(jié)果顯示,受試奶牛乳區(qū)中胰蛋白酶活性在乳房注入12 h后趨于0或者降低到檢測限以下,連續(xù)檢測24 h、36 h胰蛋白酶活性仍然低于檢測限;受試奶牛乳區(qū)中糜蛋白酶活性在乳房注入后8 h降低到檢測限以下,連續(xù)檢測12 h、24 h糜蛋白酶活性仍低于檢測限。胰蛋白酶在受試奶牛乳區(qū)中消除速度相對較慢、藥效時(shí)間長,給藥間隔可以以胰蛋白酶消除時(shí)間作為依據(jù),建議給藥間隔為12 h。為了評價(jià)復(fù)合蛋白酶乳房注入劑對臨床型奶牛乳房炎的治療效果、確定其臨床使用劑量等問題,本試驗(yàn)通過對乳房炎患牛體細(xì)胞、奶牛乳房細(xì)菌分離鑒定以及其它臨床療效指標(biāo)的檢測和觀察,評估了復(fù)合蛋白酶注入劑對臨床型奶牛乳房炎的治療效果。試驗(yàn)結(jié)果顯示,按照1支/乳區(qū)·次,連續(xù)用藥3次,每次間隔12 h給藥的臨床使用方案,復(fù)合蛋白酶乳房注入劑(糜蛋白酶8 mg+胰蛋白酶8 mg組)劑量組制劑對臨床型奶牛乳房炎的有效率達(dá)到88.2%,治愈率高達(dá)70.6%;對患病乳區(qū)病原菌菌株數(shù)清除作用減少50%以上;奶牛乳房炎評分均值從2.80提升到4.30;體細(xì)胞均值從209.80萬個(gè)/mL降低到125.94萬個(gè)/mL。建議使用主藥含量為糜蛋白酶8 mg+胰蛋白酶8 mg劑量的復(fù)合蛋白酶乳房注入劑作為臨床推薦劑量。作為一種酶制劑用做奶牛乳房炎的治療藥物,復(fù)合蛋白酶乳房注入劑為避免細(xì)菌耐藥性和藥物殘留等問題,為復(fù)合蛋白酶制劑的開發(fā)和應(yīng)用于治療奶牛乳房炎提供了新思路。為了解泌乳期奶牛對復(fù)合蛋白酶乳房注入劑的耐受程度(最大耐受劑量),明確超過推薦劑量使用復(fù)合蛋白酶乳房注入劑后可能對受試奶牛造成的包括臨床癥狀、乳房局部癥狀以及對血液、體細(xì)胞數(shù)的影響和危害,本試驗(yàn)利用不同劑量的復(fù)合蛋白酶乳房注入劑對奶牛進(jìn)行了靶動(dòng)物安全性試驗(yàn)評估。試驗(yàn)結(jié)果顯示,分別按照推薦劑量的1、3、5倍劑量為受試奶牛乳區(qū)給藥,所有受試奶牛均未觀察到明顯的不良臨床表現(xiàn)。受試奶牛血液常規(guī)和血液生化指標(biāo)在給藥前后均處在奶牛正常指標(biāo)范圍之內(nèi);分別在受試奶牛給藥的0d、3d、5 d、10 d采集奶樣進(jìn)行體細(xì)胞檢測,發(fā)現(xiàn)3×推薦劑量組、5×推薦劑量組藥物對奶牛乳房有一定程度的刺激作用,且5×推薦劑量組藥物比3×推薦劑量組藥物對奶牛乳房的刺激作用更大,建議臨床用藥時(shí)不要一次性使用超過3倍劑量的藥物;盡管3×推薦劑量和5×推薦劑量組藥物在給藥3 d和5 d后受試奶牛乳汁中的體細(xì)胞數(shù)高于50萬個(gè)/mL,但給藥10 d后乳汁中體細(xì)胞數(shù)降至20萬個(gè)/mL左右,說明該藥物會(huì)在短時(shí)間內(nèi)引起奶牛乳中體細(xì)胞數(shù)量的上升,藥效結(jié)束后,奶牛體細(xì)胞數(shù)量會(huì)降至給藥前水平,刺激強(qiáng)度可為奶牛所承受,該藥物在臨床上使用時(shí)安全性較好。
[Abstract]:Trypsin, chymotrypsin, as an enzyme drug for the treatment of inflammation, has been widely used in clinic abroad. In view of the widespread application of trypsin and chymotrypsin in human medical clinic, we intend to try to treat dairy cow mastitis with trypsin and chymotrypsin as the main component of compound protease breast injecting agent. In order to study the pharmacokinetic characteristics of the compound proteinase breast injecting agent in the cow body, the experiment of pharmacokinetics, pharmacodynamics and safety tests of the preparation in dairy cows was carried out according to the compilation of the guiding principles of veterinary medicine research technology in the Ministry of agriculture. The pharmacokinetics of the compound proteinase breast injecting agent in dairy cows (milk) was studied in this experiment. The test results showed that the lowest detection limit of trypsin in milk was 72 mu/mL, the average recovery rate was 71.9%, and the minimum detection limit of chymotrypsin was concentrated. The average recovery rate was 142 mu/mL, and the average recovery rate was 70.1%. and the test method established in this experiment was selected. The tested dairy cows were selected and 1 compound proteinase breast injections were injected into each dairy area to collect milk. The results showed that the trypsin activity in the dairy area of the dairy cows tended to 0 or lower to the detection limit after the breast injection 12 h. The activity of trypsin was still lower than the detection limit of 24 h and 36 h, and the activity of chymotrypsin decreased to below the detection limit after the breast injection of 8 h, 12 h was continuously detected, and 24 h chymotrypsin was still lower than the detection limit. It is suggested that the interval of the protease elimination is 12 h. In order to evaluate the therapeutic effect of the compound proteinase breast injecting agent on the clinical dairy cow mastitis, to determine the clinical dosage and so on. The therapeutic effect of compound proteinase injecting agent on the clinical Dairy Mastitis was evaluated. The results showed that the clinical use of the compound protease breast injecting agent (chymotrypsin 8 mg+ trypsin 8 mg group) was used for the clinical dairy cow mastitis according to the clinical use of 1 consecutive doses of 1 branches / milky areas and 12 consecutive intervals. The effective rate was 88.2%, the cure rate was up to 70.6%, the scavenging effect of the pathogenic bacteria number of the infected dairy region was reduced by more than 50%, the mean value of the cow mastitis score increased from 2.80 to 4.30, the mean of the body cell mean from 2 million 98 thousand /mL to 1 million 259 thousand and 400 /mL. suggested that the compound proteinase breasts of the main drug content was the chyloase 8 mg+ trypsin 8 mg dose. Injecting agent as a clinical recommended dose. As a kind of enzyme preparation for dairy cow mastitis, compound proteinase breast injecting agent is to avoid bacterial resistance and drug residue. It provides a new idea for the development and application of compound protease preparation for the treatment of dairy cow mastitis. The tolerance level of the breast injecting agent (maximum tolerable dose), clearly beyond the recommended dosage of the compound proteinase breast injecting agent, may include the clinical symptoms, the local symptoms of the breast, the influence and harm to the blood, the number of somatic cells, and the use of different doses of compound proteinase breast injecting agent in this experiment. The test results of the target animal safety test were conducted. The results showed that all the dairy cows were not observed the obvious adverse clinical manifestations according to the 1,3,5 times of the recommended dose, and the blood routine and blood biochemical indexes of the tested cows were within the normal range of the dairy cows before and after the Administration. The 0d, 3D, 5 d, 10 d collected milk samples were tested by the tested cows. The 3 x recommended dose group was found. The 5 x recommended dose group had a certain degree of stimulation to the cow breasts, and the 5 * recommended dose group was more stimulative to the cow breasts than the 3 x recommended dose group. It is suggested that the drug should not be used once in the clinical medication. More than 3 times the dosage of the drug; although the number of somatic cells in the milk of the dairy cows was higher than 500 thousand /mL after the 3 * recommended dose and the 5 * recommended dose group was 3 D and 5 d, the number of somatic cells in the milk was reduced to about 200 thousand /mL after the administration of 10 d, indicating that the drug could cause the increase of the number of somatic cells in the milk of dairy cows and the end of the drug effect in a short time. After that, the number of somatic cells of cows will drop to the level before administration, and the stimulation intensity can be taken by cows. The drug is safe in clinical use.
【學(xué)位授予單位】：南京農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S859.7

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