CAJ | 학술논문

目的产前对孕晚期膈疝模型雌鼠给予汉防己甲素(Tet)干预后,观察Tet对胎鼠膈疝(CDH)模型的肺血管发育和结缔组织生长因子(CTGF)的作用和影响.方法 9只健康怀孕的SD大鼠雌鼠孕9.5d被随机平均分为3组:即对照组(C组)、除草醚组(N组)和汉防己甲素组(NT组).N和NT组灌胃给予除草醚.NT组于孕第18.5～20.5天,每天给予30 mg/kg Tet灌胃,C组和N组仅给予生理盐水.孕第21.5天于麻醉下剖宫取胎鼠双肺,通过对胎肺组织行HE染色观察肺组织及血管发育,行免疫组织化学染色和RT-PCR方法观察CTGF在肺组织中的表达高低.结果本实验中N组和NT组总膈疝发生率为50％,两组间差异无统计学意义(P＞0.05).N组肺血管数目(2.5±1.1)个较C组(4.5±1.1)个有明显减少,差异有统计学意义(P＜0.01)；两组与NT组(3.7±1.4)个比较,差异没有统计学意义(P＞0.05).血管外径(ED)在各组间差异没有统计学意义(P＞0.05).血管内径(ID)表现为N组(24.8±4.7)μm＜NT组(42.7±4.0)μm＜C组(51.9±8.5)μm,各组间差异有统计学意义(P＜0.01)；血管中层(平滑肌细胞)增殖、变厚,血管中膜厚度百分比N组(58.1％±6.0％)＞NT组(25.4％±8.3％)≈C组(16.6％±9.7％),N组与另两组比较,差异有统计学意义(P＜0.01),NT组与C组间差异没有统计学意义(P＞0.05)；血管外膜厚度百分比N组(134.0％±11.9％)＞NT组(86.6％±3.8％)＞C组(66.9％±16.7％),各组组间差异有统计学意义(P＜0.01).CTGF免疫组织化学染色阳性表达强度(IOD值)结果为N组＜NT组＜C组,差异有统计学意义(P＜0.01),根据△Ct值分析基因的表达,RT-PCR结果示CTGF mRNA肺组织表达由低到高为:N组＜NT组＜C组,差异有统计学意义(P＜0.01).结论 Tet对实验大鼠动物模型的胚胎期肺血管有抑制血管重构、促血管生成和发育作用,其作用机制可能与Tet上调CTGF的表达有一定关系.
목적 산전대잉만기격산모형자서급여한방기갑소(Tet)간예후,관찰Tet대태서격산(CDH)모형적폐혈관발육화결체조직생장인자(CTGF)적작용화영향.방법 9지건강부잉적SD대서자서잉9.5d피수궤평균분위3조:즉대조조(C조)、제초미조(N조)화한방기갑소조(NT조).N화NT조관위급여제초미.NT조우잉제18.5～20.5천,매천급여30 mg/kg Tet관위,C조화N조부급여생리염수.잉제21.5천우마취하부궁취태서쌍폐,통과대태폐조직행HE염색관찰폐조직급혈관발육,행면역조직화학염색화RT-PCR방법관찰CTGF재폐조직중적표체고저.결과 본실험중N조화NT조총격산발생솔위50％,량조간차이무통계학의의(P＞0.05).N조폐혈관수목(2.5±1.1)개교C조(4.5±1.1)개유명현감소,차이유통계학의의(P＜0.01)；량조여NT조(3.7±1.4)개비교,차이몰유통계학의의(P＞0.05).혈관외경(ED)재각조간차이몰유통계학의의(P＞0.05).혈관내경(ID)표현위N조(24.8±4.7)μm＜NT조(42.7±4.0)μm＜C조(51.9±8.5)μm,각조간차이유통계학의의(P＜0.01)；혈관중층(평활기세포)증식、변후,혈관중막후도백분비N조(58.1％±6.0％)＞NT조(25.4％±8.3％)≈C조(16.6％±9.7％),N조여령량조비교,차이유통계학의의(P＜0.01),NT조여C조간차이몰유통계학의의(P＞0.05)；혈관외막후도백분비N조(134.0％±11.9％)＞NT조(86.6％±3.8％)＞C조(66.9％±16.7％),각조조간차이유통계학의의(P＜0.01).CTGF면역조직화학염색양성표체강도(IOD치)결과위N조＜NT조＜C조,차이유통계학의의(P＜0.01),근거△Ct치분석기인적표체,RT-PCR결과시CTGF mRNA폐조직표체유저도고위:N조＜NT조＜C조,차이유통계학의의(P＜0.01).결론 Tet대실험대서동물모형적배태기폐혈관유억제혈관중구、촉혈관생성화발육작용,기작용궤제가능여Tet상조CTGF적표체유일정관계.
Objective To explore the effects of prenatal tetrandrine treatment on vascular devel opment and CTGF expression in the lungs of rats with congenital diaphragmatic hernia (CDH) induced by Nitrofen.Methods Nine pregnant Sprague-Dawley rats were randomly and evenly divided into 3 groups according the treatment:C group (control group),NT group (tetrandrine group) and N group (nitrofen group).The rats of the N and NT group were administrated with 125 mg of nitrofen dissolved in 2 ml of olive oil by gastric gavage on day 9.5 of gestation,whereas those in C group only received same volume of olive oil.The rats of the NT group were administered with Tet (30 mg/kg)once a day from gestational day 18.5 to 20.5,while their counterparts of the C and N group were only treated with saline.On day 21 of gestation,all fetuses were delivered by cesarean section.The rat fetuses were sacrificed and examined for diaphragmatic hernia.Lung vascular dysplasia or remodeling was evaluated on H&E staining slides.Immunohistochcmical staining and real time polymerase chain reaction (RT-PCR) were performed to detect the expression of CTGF in lung tissues.Results Twenty-seven (50％) fetuses with CDH were harvested from the N and NT rats.No significant difference of CDH incidence was seen between the two groups (P＞0.5).More pulmonary vessels were observed in the fetuses of C group,and these vessels had thinner medial smooth muscle and adventitia,and larger lumina compared with the counterparts of N and NT group.The pulmonary vascular development of the fetuses of NT group was better than that of the N group.There were significant differences between the C and the other two groups in internal diameter (ID) (P＜0.01) and percentage of adventitial thickness (AT％) (P＜0.01).The ID of N group was smaller than that in the other 2 groups (P＜0.01),but the medial thickness (MT％) and AT％ of N group were larger than those in the other 2 groups (P＜0.01).The highest protein and mRNA expression of CTGF were observed in N group,followed by NT group and C group (P＜0.01).Conclusions Prenatal treatment with tet randrine can promote lung development and inhibit pulmonary arterial structural remodeling in fetal rats with nitrofen-induced CDH via the unregulated expression of CTGF in the lungs.