CAJ | 학술논문

目的比较不同胶原酶消化大鼠胰腺的效果,选择消化效果较好的胶原酶.方法采用随机数字表法将SD大鼠分为2组,胶原酶P组大鼠胰腺使用1 mg/ml的胶原酶P液进行消化,Ⅴ型胶原酶组大鼠胰腺使用1 mg/ml的Ⅴ型胶原酶进行消化.胰腺消化后,对获得的两组大鼠胰岛进行纯化、培养.采用双硫腙染色于倒置显微镜下计数纯化前后获得的两组全部胰岛数量,并计算胰岛当量(IEQ)；采用吖啶橙和碘丙啶双色荧光染色,于荧光显微镜下计数纯化后胰岛的活率；两组大鼠胰岛经纯化、培养2d后进行胰岛素释放试验,观察两组胰岛的功能.结果纯化前,两组间每个大鼠胰腺获得的胰岛数量的差异无统计学意义(P＞0.05)；但两组间IEQ的差异有统计学意义(P＜0.05)；经纯化后,Ⅴ型胶原酶组和胶原酶P组每个大鼠胰腺获得的胰岛数量分别为485±113和643±82,IEQ分别为674±157和989±126,两组间胰岛数量和IEQ的比较,差异均有统计学意义(P＜0.05).Ⅴ型胶原酶组和胶原酶P组大鼠胰岛经纯化后,其活率分别为(96.13±1.13)％和(96.38±0.92)％,两组比较,差异无统计学意义(P＞0.05).胰岛素释放试验结果显示,在低糖或高糖刺激下,两组间大鼠胰岛功能的差异均无统计学意义(P＞0.05).结论两种胶原酶均适用于大鼠胰腺的消化,但胶原酶P消化效果较Ⅴ型胶原酶稳定,且胰岛获得率也较高.
목적 비교불동효원매소화대서이선적효과,선택소화효과교호적효원매.방법 채용수궤수자표법장SD대서분위2조,효원매P조대서이선사용1 mg/ml적효원매P액진행소화,Ⅴ형효원매조대서이선사용1 mg/ml적Ⅴ형효원매진행소화.이선소화후,대획득적량조대서이도진행순화、배양.채용쌍류종염색우도치현미경하계수순화전후획득적량조전부이도수량,병계산이도당량(IEQ)；채용아정등화전병정쌍색형광염색,우형광현미경하계수순화후이도적활솔；량조대서이도경순화、배양2d후진행이도소석방시험,관찰량조이도적공능.결과 순화전,량조간매개대서이선획득적이도수량적차이무통계학의의(P＞0.05)；단량조간IEQ적차이유통계학의의(P＜0.05)；경순화후,Ⅴ형효원매조화효원매P조매개대서이선획득적이도수량분별위485±113화643±82,IEQ분별위674±157화989±126,량조간이도수량화IEQ적비교,차이균유통계학의의(P＜0.05).Ⅴ형효원매조화효원매P조대서이도경순화후,기활솔분별위(96.13±1.13)％화(96.38±0.92)％,량조비교,차이무통계학의의(P＞0.05).이도소석방시험결과현시,재저당혹고당자격하,량조간대서이도공능적차이균무통계학의의(P＞0.05).결론 량충효원매균괄용우대서이선적소화,단효원매P소화효과교Ⅴ형효원매은정,차이도획득솔야교고.
Objective To compare the yield rate of rats islets between different collagenase digestion groups.Methods The SD rats were randomly divided into two groups as following by using random digits table:collagenase P group (pancreas digested by 1 mg/ml collagenase P) and type Ⅴ collagenase group (pancreas digested by 1 mg/ml type Ⅴ collagenase).After pancreas digestion,rat islet cells in two groups were culture,purified and stained with DTZ.The mean islet number and islet equivalent (IEQ) before and after purification were measured under an inverted microscope.The viability of purified islets was assessed by fluorescence staining of aridine orange (AO) and propidium iodide (PI) under the fluorescence microscopy.After purification and culture for two days,islets function was evaluated by insulin releasing tests in the two groups.Results Before purification,there was no significant difference in the islets number obtained from the pancreas between two groups (P＞0.05),but there was significant difference in the IEQ (P＜0.05).After purification,the islets number in type Ⅴcollagenase group and collagenase P group was (485 ± 113)/pancrease and (643 ± 82)/pancrease,and IEQ was (674 ± 157)/pancreas and (989 ± 126)/pancreas,respectively (P＜0.05).Islet viability in type Ⅴcollagenase group and collagenase P group was (96.13 ±1.13) ％ and (96.38 ± 0.92) ％ respectively (P＞0.05).The results of insulin releasing tests revealed there was no significant difference in islet function stimulated by hypoglycemia and hyperglycemia between two groups (P＞0.05).Conclusion Two types of collagenase are suitable for the islets digestion in rats.The stability of digestion and yield rate of purified islets in collagenase P group are higher than in type Ⅴ collagenase group.