CAJ | 학술논문

目的：对人脐带间充质干细胞( mesenchymal stem cells, MSCs)在体外诱导分化为胰岛样分泌细胞进行观察,并对获得的胰岛样分泌细胞治疗效果进行评价。方法选择雄性SD大鼠32只,随机分为正常对照组(8只)、糖尿病对照组(12只)和糖尿病实验组(12只)。糖尿病对照组和糖尿病实验组给予链脲佐菌素腹腔注射进行糖尿病大鼠造模,正常对照组予0.9%氯化钠注射液腹腔注射。分离、培养脐带Wharton's Jelly(WJ)中的MSCs,将培养至第3代MSCs与雄性SD大鼠胰岛细胞共同培养14 d,培养出的干细胞移植至糖尿病实验组肾包膜下,观察56 d结束实验。通过HE染色和免疫组织化学染色观察实验结束后3组肾脏形态和细胞变化,并比较基线时与实验结束后3组血糖和体重变化。结果实验结束后正常对照组和糖尿病对照组肾脏HE染色示肾被膜边缘完整,形态清楚；免疫组织化学染色示肾被膜下无胰岛素及外源性抗原溴脱氧尿嘧啶核苷( Brdu)染色阳性的细胞。实验结束后糖尿病实验组肾脏HE染色示移植细胞部位肾被膜和肾皮质之间有大量的细胞,部分肾被膜出现断裂现象；免疫组织化学染色示肾被膜和肾皮质之间有胰岛素和Brdu阳性的细胞。基线时糖尿病实验组和糖尿病对照组血糖明显高于正常对照组,体重低于正常对照组,差异均有统计学意义(P<0．01或P<0．05)。实验结束后糖尿病实验组血糖较基线时下降,体重较基线时有所增加,差异有统计学意义(P<0．05)；3组血糖、体重比较差异均有统计学意义(P<0．05)；糖尿病实验组血糖明显低于糖尿病对照组,差异有统计学意义(P<0．05)；糖尿病实验组、糖尿病对照组体重均低于正常对照组,糖尿病实验组体重高于糖尿病对照组,差异均有统计学意义(P<0．05)。结论脐带MSCs在胰岛细胞微环境下可诱导分化为胰岛样分泌细胞,对糖尿病鼠具有降糖和恢复体重的作用。目的：對人臍帶間充質榦細胞( mesenchymal stem cells, MSCs)在體外誘導分化為胰島樣分泌細胞進行觀察,併對穫得的胰島樣分泌細胞治療效果進行評價。方法選擇雄性SD大鼠32隻,隨機分為正常對照組(8隻)、糖尿病對照組(12隻)和糖尿病實驗組(12隻)。糖尿病對照組和糖尿病實驗組給予鏈脲佐菌素腹腔註射進行糖尿病大鼠造模,正常對照組予0.9%氯化鈉註射液腹腔註射。分離、培養臍帶Wharton's Jelly(WJ)中的MSCs,將培養至第3代MSCs與雄性SD大鼠胰島細胞共同培養14 d,培養齣的榦細胞移植至糖尿病實驗組腎包膜下,觀察56 d結束實驗。通過HE染色和免疫組織化學染色觀察實驗結束後3組腎髒形態和細胞變化,併比較基線時與實驗結束後3組血糖和體重變化。結果實驗結束後正常對照組和糖尿病對照組腎髒HE染色示腎被膜邊緣完整,形態清楚；免疫組織化學染色示腎被膜下無胰島素及外源性抗原溴脫氧尿嘧啶覈苷( Brdu)染色暘性的細胞。實驗結束後糖尿病實驗組腎髒HE染色示移植細胞部位腎被膜和腎皮質之間有大量的細胞,部分腎被膜齣現斷裂現象；免疫組織化學染色示腎被膜和腎皮質之間有胰島素和Brdu暘性的細胞。基線時糖尿病實驗組和糖尿病對照組血糖明顯高于正常對照組,體重低于正常對照組,差異均有統計學意義(P<0．01或P<0．05)。實驗結束後糖尿病實驗組血糖較基線時下降,體重較基線時有所增加,差異有統計學意義(P<0．05)；3組血糖、體重比較差異均有統計學意義(P<0．05)；糖尿病實驗組血糖明顯低于糖尿病對照組,差異有統計學意義(P<0．05)；糖尿病實驗組、糖尿病對照組體重均低于正常對照組,糖尿病實驗組體重高于糖尿病對照組,差異均有統計學意義(P<0．05)。結論臍帶MSCs在胰島細胞微環境下可誘導分化為胰島樣分泌細胞,對糖尿病鼠具有降糖和恢複體重的作用。
목적：대인제대간충질간세포( mesenchymal stem cells, MSCs)재체외유도분화위이도양분비세포진행관찰,병대획득적이도양분비세포치료효과진행평개。방법선택웅성SD대서32지,수궤분위정상대조조(8지)、당뇨병대조조(12지)화당뇨병실험조(12지)。당뇨병대조조화당뇨병실험조급여련뇨좌균소복강주사진행당뇨병대서조모,정상대조조여0.9%록화납주사액복강주사。분리、배양제대Wharton's Jelly(WJ)중적MSCs,장배양지제3대MSCs여웅성SD대서이도세포공동배양14 d,배양출적간세포이식지당뇨병실험조신포막하,관찰56 d결속실험。통과HE염색화면역조직화학염색관찰실험결속후3조신장형태화세포변화,병비교기선시여실험결속후3조혈당화체중변화。결과실험결속후정상대조조화당뇨병대조조신장HE염색시신피막변연완정,형태청초；면역조직화학염색시신피막하무이도소급외원성항원추탈양뇨밀정핵감( Brdu)염색양성적세포。실험결속후당뇨병실험조신장HE염색시이식세포부위신피막화신피질지간유대량적세포,부분신피막출현단렬현상；면역조직화학염색시신피막화신피질지간유이도소화Brdu양성적세포。기선시당뇨병실험조화당뇨병대조조혈당명현고우정상대조조,체중저우정상대조조,차이균유통계학의의(P<0．01혹P<0．05)。실험결속후당뇨병실험조혈당교기선시하강,체중교기선시유소증가,차이유통계학의의(P<0．05)；3조혈당、체중비교차이균유통계학의의(P<0．05)；당뇨병실험조혈당명현저우당뇨병대조조,차이유통계학의의(P<0．05)；당뇨병실험조、당뇨병대조조체중균저우정상대조조,당뇨병실험조체중고우당뇨병대조조,차이균유통계학의의(P<0．05)。결론제대MSCs재이도세포미배경하가유도분화위이도양분비세포,대당뇨병서구유강당화회복체중적작용。
Objective To induce and observe the differentiation of islet like cells from human umbilicalcord mesen-chymal stem cell ( hUC-MSCs) in vitro, and to make evaluation of the function and effect of islet like cells diabetes. Methods 32 male SD rats were randomly divided into normal control group (8 rats), diabetic control group (12 rats) and diabetic group (12 rats). Rats in the diabetic control group and the diabetic experimental group were intraperitoneally injected with Streptozocin (STZ) injection while rats in normal control group were intraperitoneally injected with 0. 9% sodium chloride in-jection of equal volume. The hUC-MSCs form Wharton's Jelly (WJ) was isolated and the 3rd generation of hUC-MSCs was cul-tured with the rat islet cells for 14 days, and the changes of hUC-MSCs was observed under inverted microscopy, and 56 d end text as well. The islet like cells was transplanted under the renal capsule of diabetic rats of the experimental group and the changes of renal morphology and cell were observed through HE staining and immunohistochemical staining as well as the changes of the weight and blood glucose of diabetic rats during the baseline and after the experiment. Results After the ex-periment, the HE dyeing showed that the verge of renal capsule was integrity and clear in normal control group and diabetic control group. The immunohistochemistry dyeing showed that there were not positive cells for insulin or Brdu below the renal capsule in normal control group and diabetic control group. There were considerable cells between renal capsule and renal cor-tex in the diabetic experimental group. The renal capsule appeared to be broken. There were some positive cells for insulin whose endochylemas were brown as well as some positive cells for Brdu whose nucelus were brown between renal capsule and renal cortex in the diabetic experimental group. At the baseline, the blood glucose of the diabetic experimental group and the diabetic control group were significantly higher than those of the normal control group while the weight was lower with statisti-cally significant difference ( P <0. 01 or P <0. 05 ) . After the experiment, the blood glucose of the diabetic experimental group was decreased compared with baseline, but the weight increased, and the difference was statistically significant ( P<0. 05), difference of blood glucose and weight between the 3 groups were statistically significant (P<0. 05). The blood glu-cose of diabetic experimental group was lower than that of the diabetes control group, and the difference was statistically signif-icant (P<0. 05). The weight of diabetic experimental group and diabetic control group was lower than that of the normal con-trol group, while the weight of diabetic experimental group was higher than that of the diabetes control group, and the differ-ence was statistically significant (P<0. 05). Conclusion In the islet cell microenvironment, the umbilical cord MSCs can be induced and differentiated into islet like cells, which can reduce blood glucose level and rebalance the weight of diabetic patients.