CAJ | 학술논문

链脲佐菌素法建立糖尿病大鼠模型.分为正常组、糖尿病组和漆黄素干预组.24周后处死大鼠,检测血,尿标本相关生化指标.石蜡切片HE染色观察病理改变,PAS、Masson染色分析细胞外基质表达.免疫组化检测p300表达,Western印迹法检测p300及基质金属蛋白酶2(MMP-2)蛋白水平.实时定量PCR法分析MMP-2 mRNA表达.与糖尿病组相比较,漆黄素干预组生化指标显著改善,肾脏病理改变减轻；进一步染色分析示细胞外基质蛋白表达较糖尿病组显著降低；但较正常组增高.免疫组化分析,3组模型的p300蛋白主要在肾脏肾小球表达,且在细胞核,细胞质均有表达.而经漆黄素干预后p300蛋白着色较糖尿病组明显变浅.Western印迹定量分析发现漆黄素干预组的p300蛋白表达较糖尿病组显著下调；漆黄素干预组MMP-2 mRNA及蛋白表达均较糖尿病组增高,但较正常组降低,差异有统计学意义(P＜0.05).提示漆黄素可显著改善糖尿病大鼠肾脏病变,此可能与其抑制糖尿病诱导的p300表达和促进MMP-2表达有关.
련뇨좌균소법건립당뇨병대서모형.분위정상조、당뇨병조화칠황소간예조.24주후처사대서,검측혈,뇨표본상관생화지표.석사절편HE염색관찰병리개변,PAS、Masson염색분석세포외기질표체.면역조화검측p300표체,Western인적법검측p300급기질금속단백매2(MMP-2)단백수평.실시정량PCR법분석MMP-2 mRNA표체.여당뇨병조상비교,칠황소간예조생화지표현저개선,신장병리개변감경；진일보염색분석시세포외기질단백표체교당뇨병조현저강저；단교정상조증고.면역조화분석,3조모형적p300단백주요재신장신소구표체,차재세포핵,세포질균유표체.이경칠황소간예후p300단백착색교당뇨병조명현변천.Western인적정량분석발현칠황소간예조적p300단백표체교당뇨병조현저하조；칠황소간예조MMP-2 mRNA급단백표체균교당뇨병조증고,단교정상조강저,차이유통계학의의(P＜0.05).제시칠황소가현저개선당뇨병대서신장병변,차가능여기억제당뇨병유도적p300표체화촉진MMP-2표체유관.
Rat models of diabetes were established by injecting streptozocin intraperitoneally.According to random number table,three groups were divided:normal group,diabetic group,and fisetin-treated group.After 24 weeks,all rats were sacrificed.Biochemical parameters of blood and urine samples were tested.The pathological changes were observed by paraffin sections staining with HE.The expression of extracellular matrix proteins was analyzed via PAS and Masson staining.Location of p300 protein expression was analyzed by immunohistochemistry.The protein expressions of p300 and MMP-2 were determined by Western blotting.The mRNA expressions of MMP-2 were analyzed via real-time PCR.The biochemical parameters and kidney pathological images in fisetin-treated group were better than those in diabetic group.The expression of extraeellular matrix proteins was lower than that in diabetic group.Immunohistochemistry analysis showed that among three groups the expression of p300 was mainly in glomeruli,and was also expressed in cell nucleus and cytoplasm and the coloration of fisetin-treated group was weakened as compared with diabetic group.Western blotting analysis showed that the expression of p300 protein in fisetin-treated group was lower than that in diabetic group(P＜O.05).The expressions of MMP-2 mRNA and MMP-2 protein were higher than those in diabetic group (P ＜ 0.05).It is suggested that fisetin may attenuate diabetes associated abnormalities in the kidney of rats,owing probably to inhibiting the expressions of p300 and enhancing the expressions of MMP-2.