国际呼吸杂志
國際呼吸雜誌
국제호흡잡지
INTERNATIONAL JOURNAL OF RESPIRATION
2015年
5期
351-357
,共7页
肺气肿大鼠%骨髓间充质干细胞%转化生长因子β1%肿瘤坏死因子α
肺氣腫大鼠%骨髓間充質榦細胞%轉化生長因子β1%腫瘤壞死因子α
폐기종대서%골수간충질간세포%전화생장인자β1%종류배사인자α
Emphysema rat%Bone marrow mesenchymal stem cells%Transforming growth factor-β1%Tumor necrosis factor-α
目的 通过研究骨髓间充质干细胞(BMSCs)对肺气肿大鼠肺组织和血清中转化生长因子β1(TGF-β1)和肿瘤坏死因子α(TNF-α)表达的影响,探讨BMSCs移植对肺气肿大鼠气道炎症和重塑的影响.方法 健康雄性SD大鼠36只,随机分为对照组、单纯肺气肿组和BMSCs移植组.后两组参照许三林慢性烟雾暴露装置建立肺气肿大鼠模型,慢性烟雾暴露6个月后,给予BMSCs移植组大鼠尾静脉注射BMSCs,单纯肺气肿组大鼠尾静脉注射等量的PBS缓冲液,观察BMSCs移植后各组大鼠肺组织病理学变化,并定量测定肺组织平均内衬间隔(MLI)、单位面积平均肺泡数(MAN)和管壁总面积(WAt)/管壁内周长(Pi);采用ELISA方法测定肺组织匀浆和血清TGF-β1和TNF-α的水平,免疫组织化学半定量法测定气道上皮细胞TGF-β1的表达水平.结果 ①镜下形态学改变:单纯肺气肿组和BMSCs移植组都存在不同程度的肺气肿病理改变,后者肺气肿严重程度较前者为轻;②单纯肺气肿组MLI(177±22.14) μm和WAt/Pi(193.05±53.91) μm、肺组织及血清TNF-α[(14.47±5.85)及(92.07±23.73) ng/L]、TGF-β1[(66.77±9.91)和(72.92±4.96)μg/L]和气道上皮细胞TGF-β1阳性颗粒的平均光密度值(6.44±2.40)×10-2明显高于对照组[(95±44.28)μm和(78.39±26.51) μm、(5.17±2.57)和(53.49±15.47) μg/L、(49.02±9.91)和(59.36±6.89) μg/L和(3.83±1.81)×10-2,P值均＜0.05],MAN(92.73±28.93)/mm2明显低于对照组[(190.84±57.49)/mm2,P＜0.05)],差异均有统计学意义;BMSCs移植组MLI(131±37.55) μm和WAt/Pi (121.62±34.94) μm、肺组织及血清TNF-α[(9.60±5.06)和(70.92±15.16)] ng/L、TGF-β1[(57.44±4.78)和(66.43±6.70) μg/L]、气道上皮细胞TGF-β1阳性颗粒的平均光密度值(4.28±1.23)×10-2均明显高于对照组(P＜0.05),MAN (146.73±49.12)]/mm2明显低于对照组(P＜0.05),差异均有统计学意义;而BMSCs移植组MLI、WAt/Pi、TNF-α及TGF-β1水平均明显低于单纯肺气肿组(P值均＜0.05),MAN明显高于单纯肺气肿组(P＜0.05),3组比较差异均有统计学意义(F值为3.845～27.506,P＜0.05或P＜0.01);③各组间大鼠TGF-β1与TNF-α水平(r=0.556,P=0.001)及WAt(r =0.702,P＜0.01)均呈正相关;TNF-α水平与MLI(r =0.578,P=0.001)及WAt(r=0.440,P=0.015)均呈正相关,与MAN(r=-0.439,P=0.015)呈负相关;WAt与MLI(r=0.398,P=0.029)呈正相关,与MAN(r=-0.475,P=0.008)呈负相关.结论 BMSCs移植可降低肺气肿大鼠肺组织和血清TGF-β1和TNF-α的表达水平,减轻气道炎症和气道重塑,进而改善其肺气肿严重程度.
目的 通過研究骨髓間充質榦細胞(BMSCs)對肺氣腫大鼠肺組織和血清中轉化生長因子β1(TGF-β1)和腫瘤壞死因子α(TNF-α)錶達的影響,探討BMSCs移植對肺氣腫大鼠氣道炎癥和重塑的影響.方法 健康雄性SD大鼠36隻,隨機分為對照組、單純肺氣腫組和BMSCs移植組.後兩組參照許三林慢性煙霧暴露裝置建立肺氣腫大鼠模型,慢性煙霧暴露6箇月後,給予BMSCs移植組大鼠尾靜脈註射BMSCs,單純肺氣腫組大鼠尾靜脈註射等量的PBS緩遲液,觀察BMSCs移植後各組大鼠肺組織病理學變化,併定量測定肺組織平均內襯間隔(MLI)、單位麵積平均肺泡數(MAN)和管壁總麵積(WAt)/管壁內週長(Pi);採用ELISA方法測定肺組織勻漿和血清TGF-β1和TNF-α的水平,免疫組織化學半定量法測定氣道上皮細胞TGF-β1的錶達水平.結果 ①鏡下形態學改變:單純肺氣腫組和BMSCs移植組都存在不同程度的肺氣腫病理改變,後者肺氣腫嚴重程度較前者為輕;②單純肺氣腫組MLI(177±22.14) μm和WAt/Pi(193.05±53.91) μm、肺組織及血清TNF-α[(14.47±5.85)及(92.07±23.73) ng/L]、TGF-β1[(66.77±9.91)和(72.92±4.96)μg/L]和氣道上皮細胞TGF-β1暘性顆粒的平均光密度值(6.44±2.40)×10-2明顯高于對照組[(95±44.28)μm和(78.39±26.51) μm、(5.17±2.57)和(53.49±15.47) μg/L、(49.02±9.91)和(59.36±6.89) μg/L和(3.83±1.81)×10-2,P值均＜0.05],MAN(92.73±28.93)/mm2明顯低于對照組[(190.84±57.49)/mm2,P＜0.05)],差異均有統計學意義;BMSCs移植組MLI(131±37.55) μm和WAt/Pi (121.62±34.94) μm、肺組織及血清TNF-α[(9.60±5.06)和(70.92±15.16)] ng/L、TGF-β1[(57.44±4.78)和(66.43±6.70) μg/L]、氣道上皮細胞TGF-β1暘性顆粒的平均光密度值(4.28±1.23)×10-2均明顯高于對照組(P＜0.05),MAN (146.73±49.12)]/mm2明顯低于對照組(P＜0.05),差異均有統計學意義;而BMSCs移植組MLI、WAt/Pi、TNF-α及TGF-β1水平均明顯低于單純肺氣腫組(P值均＜0.05),MAN明顯高于單純肺氣腫組(P＜0.05),3組比較差異均有統計學意義(F值為3.845～27.506,P＜0.05或P＜0.01);③各組間大鼠TGF-β1與TNF-α水平(r=0.556,P=0.001)及WAt(r =0.702,P＜0.01)均呈正相關;TNF-α水平與MLI(r =0.578,P=0.001)及WAt(r=0.440,P=0.015)均呈正相關,與MAN(r=-0.439,P=0.015)呈負相關;WAt與MLI(r=0.398,P=0.029)呈正相關,與MAN(r=-0.475,P=0.008)呈負相關.結論 BMSCs移植可降低肺氣腫大鼠肺組織和血清TGF-β1和TNF-α的錶達水平,減輕氣道炎癥和氣道重塑,進而改善其肺氣腫嚴重程度.
목적 통과연구골수간충질간세포(BMSCs)대폐기종대서폐조직화혈청중전화생장인자β1(TGF-β1)화종류배사인자α(TNF-α)표체적영향,탐토BMSCs이식대폐기종대서기도염증화중소적영향.방법 건강웅성SD대서36지,수궤분위대조조、단순폐기종조화BMSCs이식조.후량조삼조허삼림만성연무폭로장치건립폐기종대서모형,만성연무폭로6개월후,급여BMSCs이식조대서미정맥주사BMSCs,단순폐기종조대서미정맥주사등량적PBS완충액,관찰BMSCs이식후각조대서폐조직병이학변화,병정량측정폐조직평균내츤간격(MLI)、단위면적평균폐포수(MAN)화관벽총면적(WAt)/관벽내주장(Pi);채용ELISA방법측정폐조직균장화혈청TGF-β1화TNF-α적수평,면역조직화학반정량법측정기도상피세포TGF-β1적표체수평.결과 ①경하형태학개변:단순폐기종조화BMSCs이식조도존재불동정도적폐기종병리개변,후자폐기종엄중정도교전자위경;②단순폐기종조MLI(177±22.14) μm화WAt/Pi(193.05±53.91) μm、폐조직급혈청TNF-α[(14.47±5.85)급(92.07±23.73) ng/L]、TGF-β1[(66.77±9.91)화(72.92±4.96)μg/L]화기도상피세포TGF-β1양성과립적평균광밀도치(6.44±2.40)×10-2명현고우대조조[(95±44.28)μm화(78.39±26.51) μm、(5.17±2.57)화(53.49±15.47) μg/L、(49.02±9.91)화(59.36±6.89) μg/L화(3.83±1.81)×10-2,P치균＜0.05],MAN(92.73±28.93)/mm2명현저우대조조[(190.84±57.49)/mm2,P＜0.05)],차이균유통계학의의;BMSCs이식조MLI(131±37.55) μm화WAt/Pi (121.62±34.94) μm、폐조직급혈청TNF-α[(9.60±5.06)화(70.92±15.16)] ng/L、TGF-β1[(57.44±4.78)화(66.43±6.70) μg/L]、기도상피세포TGF-β1양성과립적평균광밀도치(4.28±1.23)×10-2균명현고우대조조(P＜0.05),MAN (146.73±49.12)]/mm2명현저우대조조(P＜0.05),차이균유통계학의의;이BMSCs이식조MLI、WAt/Pi、TNF-α급TGF-β1수평균명현저우단순폐기종조(P치균＜0.05),MAN명현고우단순폐기종조(P＜0.05),3조비교차이균유통계학의의(F치위3.845～27.506,P＜0.05혹P＜0.01);③각조간대서TGF-β1여TNF-α수평(r=0.556,P=0.001)급WAt(r =0.702,P＜0.01)균정정상관;TNF-α수평여MLI(r =0.578,P=0.001)급WAt(r=0.440,P=0.015)균정정상관,여MAN(r=-0.439,P=0.015)정부상관;WAt여MLI(r=0.398,P=0.029)정정상관,여MAN(r=-0.475,P=0.008)정부상관.결론 BMSCs이식가강저폐기종대서폐조직화혈청TGF-β1화TNF-α적표체수평,감경기도염증화기도중소,진이개선기폐기종엄중정도.
Objective By research the expression of TGF-β1 and TNF-α level in lung tissue and serum after transplantation of bone marrow mesenchymal stem cells (BMSCs) to emphysema rats,study the effects of BMSCs on airway inflammation and remodeling of emphysema rats.Methods Thirty-six healthy male SD rats were randomly divided into control group,simple emphysema and BMSCs transplantation group.Rat models of emphysema were established by exposure to chronic tobacco smoke with reference to Xu Sanlin chronic smoke exposure device,six months later,the BMSCs suspension was infused into the tail vein of BMSCs translation group rats,and the equal amount of PBS buffer was infused into the control group.The relevant indices were measured after BMSCs transplantation,the pathology changes of lung tissue were observed,and the mean linear intercept (MLI),mean alveoli number (MAN) and total area of the tube wall (WAt)/wall inside the perimeter (Pi) were calculated after HE staining,the levels of TGF-β1 and TNF-α in lung tissue and serumwere detected by ELISA kits,the TGF-β1 expression of airway epithelial cells was measured by immunohistochemical method.Results ① The microscopic morphology change.:pure emphysema and BMSCs transplantation group both have different degrees of emphysema pathological changes,the latter is significantly lighter than the former.②The MLI (177± 22.14) μm and WAt/Pi (198.05 ± 53.91) μm in the simple emphysema group were obviously higher than that of control group [(95±44.28)μm and (78.39±26.51) μm,allP ＜0.05).The TNF-α [(14.47±5.85) and (92.07±23.73)] ng/L and TGF-β1 [(66.77 ± 9.91) and (72.92±4.96)] μg/L levels in lung tissue and serum and the TGF-β1 average optical density value of the positive particle in airway epithelial cell (6.44±2.40) × 10-2 of the simple emphysema group were obviously higher than that of control group [(5.17±2.57) and (53.49 ± 15.47) ng/L,(49.02 ± 9.91) and (59.36 ± 6.89) μg/L and (3.83 ± 1.81)× 10-2,all P ＜0.05],the MAN (92.73 ± 28.93)/mm2 was obviously lower than that of control group [(190.84±57.49)/mm2,P ＜0.05)].The MLI(131±37.55) μm and WAt/Pi (121.62± 34.94) μm in the BMSCs transplantation group were obviously higher than that of control group.The TNF-α [(9.60±5.06) and (70.92±15.16) ng/L],TGF-β11 [(57.44±4.78) and (66.43±6.70)] μg/L levels in lung tissue and serum and the TGF-β1 average optical density value of the positive particle in airway epithelial cell (4.28± 1.23) × 10-2 in the BMSCs transplantation group were obviously higher than that of control group (P ＜ 0.05),the MAN (146.73 ± 49.12)/xm2 was obviously lower than that of control group (P ＜ 0.05),while the BMSCs transplantation group were obviously lower than simple emphysema (P ＜0.05).③In each group rats,the lung tissue and serum TGF-β1 level correlated positively with TNF-α level (r =0.556,P =0.001)and WAt (r =0.702,P ＜0.01).The lung tissue and serum TNF-α level correlated positively with MLI(r =0.578,P =0.001) and WAt (r =0.440,P =0.015),and correlated negatively with MAN (r =-0.439,P =0.015).WAt was positively correlated with MLI(r =0.398,P =0.029),and negatively correlated with MAN (r =-0.475,P =0.008).Conclusions Transplantation of BMSCs can reduce the TGF-β1 and TNF-α levels in lung tissue and serum of emphysema rat,then reduce airway inflammation and remodeling,and thus improve the severity of emphysema.
