CAJ | 학술논문

目的：探讨甲泼尼龙琥珀酸钠对自身免疫性肺气肿大鼠肺泡隔细胞凋亡的影响及作用机制，为研究慢性阻塞肺疾病（chronic obstructive pulmonary disease，COPD）发病机制及寻求新的治疗方法提供理论依据。方法24只SD大鼠随机分为3组：佐剂对照组（n＝8）、自身免疫性肺气肿模型组（简称模型组，n＝8）及甲泼尼龙琥珀酸钠干预组（简称干预组，n＝8）。将原代培养的人脐静脉内皮细胞及佐剂注入模型组大鼠腹腔内建立自身免疫性肺气肿模型，干预组同时腹腔内注入注射用甲泼尼龙琥珀酸钠10 mg/（kg·d）的剂量进行干预。佐剂对照组腹腔内只注射等量的佐剂。21 d后处死所有大鼠。每组均取肺组织切片进行苏木精-伊红（HE）染色观察病理形态学改变，并定量测定肺平均内衬间隔（mean linear intercept ，MLI）、平均肺泡数（mean alveolar numbers ， MAN），利用免疫组化法测定血管内皮生长因子（vascular endothelial growth factor ，VEGF）、血管内皮细胞生长因子-2（vascular endothelial growth factor-recptor2，VEGFR-2）在肺内的表达；采用脱氧核糖核酸末端转移酶介导的 dUTP 缺口末端标记 Terminal deoxynucleotidyl transferasemediated dUTP nick end labeling ，TUNEL）技术检测肺泡隔细胞调亡。结果模型组MLI比佐剂对照组明显增高（P＜0.05），但MAN比佐剂对照组明显降低（P＜0.05）；干预组MLI比模型组明显降低（P＜0.05），但MAN比模型组明显增高（P＜0.05）。模型组肺内VEGF、VEGFR-2的表达较佐剂对照组明显降低，干预组肺内VEGF、VEGFR-2的表达较模型组明显升高，差异均有统计学意义（P＜0.05）。模型组肺泡间隔细胞凋亡指数（apoptosis index，AI）明显高于佐剂对照组；干预组AI低于模型组，但高于佐剂对照组，差异均有统计学意义（P＜0.05）。结论肺组织内VEGF、VEGFR-2表达下降可能通过促进肺泡隔细胞凋亡，参与大鼠自身免疫性肺气肿的形成；甲泼尼龙琥珀酸钠可缓解自身免疫性肺气肿的形成，其机制可能与调控VEGF、VEGFR-2表达及抑制肺泡间隔细胞凋亡有关。目的：探討甲潑尼龍琥珀痠鈉對自身免疫性肺氣腫大鼠肺泡隔細胞凋亡的影響及作用機製，為研究慢性阻塞肺疾病（chronic obstructive pulmonary disease，COPD）髮病機製及尋求新的治療方法提供理論依據。方法24隻SD大鼠隨機分為3組：佐劑對照組（n＝8）、自身免疫性肺氣腫模型組（簡稱模型組，n＝8）及甲潑尼龍琥珀痠鈉榦預組（簡稱榦預組，n＝8）。將原代培養的人臍靜脈內皮細胞及佐劑註入模型組大鼠腹腔內建立自身免疫性肺氣腫模型，榦預組同時腹腔內註入註射用甲潑尼龍琥珀痠鈉10 mg/（kg·d）的劑量進行榦預。佐劑對照組腹腔內隻註射等量的佐劑。21 d後處死所有大鼠。每組均取肺組織切片進行囌木精-伊紅（HE）染色觀察病理形態學改變，併定量測定肺平均內襯間隔（mean linear intercept ，MLI）、平均肺泡數（mean alveolar numbers ， MAN），利用免疫組化法測定血管內皮生長因子（vascular endothelial growth factor ，VEGF）、血管內皮細胞生長因子-2（vascular endothelial growth factor-recptor2，VEGFR-2）在肺內的錶達；採用脫氧覈糖覈痠末耑轉移酶介導的 dUTP 缺口末耑標記 Terminal deoxynucleotidyl transferasemediated dUTP nick end labeling ，TUNEL）技術檢測肺泡隔細胞調亡。結果模型組MLI比佐劑對照組明顯增高（P＜0.05），但MAN比佐劑對照組明顯降低（P＜0.05）；榦預組MLI比模型組明顯降低（P＜0.05），但MAN比模型組明顯增高（P＜0.05）。模型組肺內VEGF、VEGFR-2的錶達較佐劑對照組明顯降低，榦預組肺內VEGF、VEGFR-2的錶達較模型組明顯升高，差異均有統計學意義（P＜0.05）。模型組肺泡間隔細胞凋亡指數（apoptosis index，AI）明顯高于佐劑對照組；榦預組AI低于模型組，但高于佐劑對照組，差異均有統計學意義（P＜0.05）。結論肺組織內VEGF、VEGFR-2錶達下降可能通過促進肺泡隔細胞凋亡，參與大鼠自身免疫性肺氣腫的形成；甲潑尼龍琥珀痠鈉可緩解自身免疫性肺氣腫的形成，其機製可能與調控VEGF、VEGFR-2錶達及抑製肺泡間隔細胞凋亡有關。
목적：탐토갑발니룡호박산납대자신면역성폐기종대서폐포격세포조망적영향급작용궤제，위연구만성조새폐질병（chronic obstructive pulmonary disease，COPD）발병궤제급심구신적치료방법제공이론의거。방법24지SD대서수궤분위3조：좌제대조조（n＝8）、자신면역성폐기종모형조（간칭모형조，n＝8）급갑발니룡호박산납간예조（간칭간예조，n＝8）。장원대배양적인제정맥내피세포급좌제주입모형조대서복강내건립자신면역성폐기종모형，간예조동시복강내주입주사용갑발니룡호박산납10 mg/（kg·d）적제량진행간예。좌제대조조복강내지주사등량적좌제。21 d후처사소유대서。매조균취폐조직절편진행소목정-이홍（HE）염색관찰병리형태학개변，병정량측정폐평균내츤간격（mean linear intercept ，MLI）、평균폐포수（mean alveolar numbers ， MAN），이용면역조화법측정혈관내피생장인자（vascular endothelial growth factor ，VEGF）、혈관내피세포생장인자-2（vascular endothelial growth factor-recptor2，VEGFR-2）재폐내적표체；채용탈양핵당핵산말단전이매개도적 dUTP 결구말단표기 Terminal deoxynucleotidyl transferasemediated dUTP nick end labeling ，TUNEL）기술검측폐포격세포조망。결과모형조MLI비좌제대조조명현증고（P＜0.05），단MAN비좌제대조조명현강저（P＜0.05）；간예조MLI비모형조명현강저（P＜0.05），단MAN비모형조명현증고（P＜0.05）。모형조폐내VEGF、VEGFR-2적표체교좌제대조조명현강저，간예조폐내VEGF、VEGFR-2적표체교모형조명현승고，차이균유통계학의의（P＜0.05）。모형조폐포간격세포조망지수（apoptosis index，AI）명현고우좌제대조조；간예조AI저우모형조，단고우좌제대조조，차이균유통계학의의（P＜0.05）。결론폐조직내VEGF、VEGFR-2표체하강가능통과촉진폐포격세포조망，삼여대서자신면역성폐기종적형성；갑발니룡호박산납가완해자신면역성폐기종적형성，기궤제가능여조공VEGF、VEGFR-2표체급억제폐포간격세포조망유관。
Objective To investigate the effects of methyprednislone on the expression of tumor alveolar septal cell apoptosis in autoimmune emphysema rats,in order to provide a theoretical basis for Chronic obstructive pulmonary disease(COPD)pathogenesis and it’s treatment.Methods 24 rats were randomly divided into three groups:normal control group (n=8 ),model group (n=8 )and intervention group (methylprednisolone sodium succinate,n=8).Intraperitoneal injection of primary cultured human umbilical vein endothelial cells were given to establish the autoimmune emphysema models,intervention group was injected with methylprednisolone at the meantime,and normal control group was received adjuvant only.Pathological changes were observed in lung tissues stained by hematoxylin eosin,mean liner intercept(MLI)and mean alveolar numbers(MAN)were measured.The localization of VEGF and VEGFR-2 in lung tissues were detected by immunohistochemical analysis.Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)technique was carried out to detect the alveolar septal cells apoptosis. Results The MLI in model group was higher than that in normal control group,while MAN was lower(P<0.05 );MLI in intervention group was lower than that in model group,but the MAN was higher (P<0.05 ).The localization of VEGF and VEGFR-2 in lung tissues in model group were lower than that in normal control group,and those in intervention group were higher than that in model group,the difference were all statistically significant(P<0.05 ).AI of alveolar septal cell in model group was higher than that in normal control group,which in intervention group was higher than that in model group,the difference were all statistically significant(P<0.05).Conclusion The decreasing of VEGF and VEGFR-2 in lung tissues may cause alveolar septal cell apoptosis and contribute to the pathogenesis of autoimmune emphysema of rats.Methyprednislone can alleviate the form of autoimmune emphysema in rats,which may be ralated to the regulation of VEGF and VEGFR-2 expression and inhibition of alveolar septal cell apoptosis.