【摘要】：目的: 探討脂肪間充質(zhì)干細(xì)胞(ADMSC)移植治療對(duì)野百合堿誘發(fā)的大鼠肺動(dòng)脈高壓肺血管重構(gòu)的影響。 方法: 取SD大鼠腹股溝皮下脂肪,膠原酶消化法分離培養(yǎng)ADMSC,移植前用帶GFP的腺病毒轉(zhuǎn)染標(biāo)記。雄性Sprague-Dawley大鼠90只,隨機(jī)均分為三組,每組30只:正常對(duì)照組(Ctr)、肺動(dòng)脈高壓組(PAH)、脂肪間充質(zhì)干細(xì)胞治療組(ADMSC)。PAH組和ADMSC組一次性腹腔注射MCT40mg/Kg,Ctr組經(jīng)腹腔注射相同體積的生理鹽水,兩周后ADMSC組經(jīng)左頸外靜脈一次性給予ADMSC 10~6/1ml,Ctr組和PAH組經(jīng)左頸外靜脈一次性給予1ml的生理鹽水。分別于ADMSC移植后1W、2W、3W,右心導(dǎo)管法測(cè)定大鼠的肺動(dòng)脈平均壓(MPAP)、右心室肥厚指數(shù)[RVHI%=右心室游離壁(RV)重量/左心室+室間隔(LV+S)重量]、并運(yùn)用ipp6.0圖像分析軟件分析肺組織切片,測(cè)定直徑在100-200μm范圍的肺小動(dòng)脈管壁厚度(WT)占動(dòng)脈外徑(ED)的百分比(WT%)及管壁面積(WA)占血管總面積的百分比(WA%)。熒光顯微鏡觀察帶GFP標(biāo)記的ADMSC在肺內(nèi)的定植。 結(jié)果: (1)ADMSC治療后1W,2W,3W,三個(gè)時(shí)間點(diǎn),每一個(gè)時(shí)間點(diǎn),PAH組大鼠與正常組相比,大鼠的MPAP和RVHI%均高于Ctr組[MPAP(mmHg): (24.53±2.90) VS (14.81±1.95),(33.18±2.30)VS(15.08±2.35),(36.38±3.28)VS(15.65±2.27);RVHI%:(41.01±1.29)VS(28.25±2.15),(48.75±2.13)VS(28.71±2.08),(50.52±1.49)VS(28.54±2.19);P均0.05]。而經(jīng)ADMSC治療后,MPAP和RVHI%較PAH組下降[MPAP(mmHg): (18.63±2.15) VS(24.53±2.90),(23.07±2.84)VS(33.18±2.30),(22.98±2.34)VS(36.38±3.28);RVHI%:(36.21±4.27)VS(41.01±1.29),(39.47±4.02)VS(48.75±2.13), (41.02±0.93)VS(50.52±1.49);P均0.05]。 (2)ADMSC治療后1W,2W,3W,三個(gè)時(shí)間點(diǎn),每一個(gè)時(shí)間點(diǎn),PAH組大鼠與正常組相比,大鼠的WA%和WT%均明顯高于Ctr組[WA%: (68.70±3.43) VS(50.27±4.92),(80.48±6.19)VS(51.61±3.29),(84.01±2.76)VS(52.02±5.87);WT%:(46.09±4.70)VS(32.46±5.19),(57.26±4.32)VS(32.52±3.19),(64.64±3.86)VS(31.42±4.53);P均0.05]。而經(jīng)ADMSC治療后,WA%和WT%均較PAH組下降[WA%:(58.23±4.08) VS(68.70±3.43),(62.97±6.58)VS(80.48±6.19),(65.27±5.45)VS(84.01±2.76);WT%:(38.06±4.15)VS(46.09±4.70),(40.91±5.24)VS(57.26±4.32), (41.91±5.16)VS(64.64±3.86);P均0.05]。 (3)與ADMSC治療后1W比,ADMSC治療后2W、3W,治療組大鼠的MPAP、RVHI%、WA %和WT %均高[MPAP ( mmHg ):( 23.07±2.84 )、( 22.98±2.34 ) VS (18.63±2.15);RVHI%:(39.47±4.02)、(41.02±0.93)VS(36.21±4.27);WA%:(62.97±6.58)、(65.27±5.45)VS (58.23±4.08) ;WT%:(40.91±5.24)、(41.91±5.16)VS(38.06±4.15);P均0.05]。ADMSC治療后2W與ADMSC治療后3W相比,治療組大鼠的MPAP、RVHI%、WA%和WT%沒(méi)有統(tǒng)計(jì)學(xué)意義[MPAP(mmHg):(23.07±2.84)VS(22.98±2.34);RVHI%:(39.47±4.02)VS(41.02±0.93);WA%:(62.97±6.58)VS(65.27±5.45);WT%:(40.91±5.24)VS(41.91±5.16);P均0.05]。 (4)熒光顯微鏡觀察到GFP標(biāo)記的ADMSC在肺內(nèi)的定植。 結(jié)論:ADMSC可以有效地降低野百合堿誘發(fā)的肺動(dòng)脈高壓大鼠的肺動(dòng)脈壓力,減輕右心室肥厚,改善肺小動(dòng)脈的重構(gòu);ADMSC治療1W時(shí),僅是延緩MCT誘導(dǎo)的肺動(dòng)脈高壓的進(jìn)展,而ADMSC治療2W后,其治療作用達(dá)到穩(wěn)定的水平。
[Abstract]:Objective:
Objective To investigate the effects of adipose-derived mesenchymal stem cells (ADMSC) transplantation on pulmonary vascular remodeling in rats with monocrotaline-induced pulmonary hypertension.
Method:
90 Sprague-Dawley male rats were randomly divided into three groups: normal control group (Ctr), pulmonary hypertension group (PAH), adipose-derived mesenchymal stem cell treatment group (ADMSC), PAH group and ADMSC group. The same volume of normal saline was injected intraperitoneally in the Ctr group. The ADMSC group was given 10-6/1ml once through the left external jugular vein two weeks later. The Ctr group and PAH group were given 1 ml of normal saline once through the left external jugular vein. The index [RVHI% = RV weight / LV + S weight] was used to analyze lung tissue slices. The percentage of pulmonary arterial wall thickness (WT) to arterial diameter (ED) and the percentage of wall area (WA) to total vascular area (WA%) were measured by fluorescence microscopy. Observe the colonization of ADMSC labeled with GFP in the lung.
Result:
(1) The MPAP and RVHI% of PAH and RVHIin PAH group were significantly higher than those of Ctr group [MPAP (mmHg): (24.53 ((24.53 ((2.90) VS (14.81 ((14.81 ((24.53 (2.90) VS (14.81 (1.95), (33.18 (2.30) VS (15.08 (15.08 (15.08 (15.08 (15.35), (36.38 (3.28) VS (15.65 (15.65 (2.27); RVHIV%: (41.01 (1.01 (1.29) VS (24.29) VS (28.29) 28 VS (28.71 +2.08), (50.5) After ADMSC treatment, MPAP (mmHg) was (18.63 ((mmHg): (18.63 ((2.15) VS (24.53 ((24.53 (2.90), (23.07 ((2.84) VS (33.18 (2.30), (22.98 (2.34) VS (36.38 (3.38 (3.28); (22.38 (3.28); (22.98 (2.38 (3.28); (36.21 ((36.21 (4.27) VS (41.01 (41.01 (41.01), (41.01 (1 1 1.01 (24.01 (24.01 (24.29 48.75 (+ 2.13), 41.02 (+ 0.93) VS (50.52 (+ 1.49), P 0.05.
(2) After ADMSC treatment, 1 W, 2, 2, 2, 3 W, three time points, at each time point, the WA% and WT% of PAH group rats were significantly higher than those of Ctr group [WA%: (68.70 ((68.70 (3.43) VS (50.27 [(50.27 [(50.27 [(50.92)%, (50.27 [(50.27 [80.47 [(68.70 [(68.70 (3.43) 43) VS (50.27 (50.27 [80.47 [80.48 [6.19] 6.19) VS (51.61 [80.48 [6.48 [6.19] 19] VS (51.61 [52 + 3.19), (64.64 + 3.86) After treatment with ADMSC, both WA% and WT% were significantly lower than PAH group [WA%: (58.23 [(58.23 [(58.23 [(4.08) VS (68.70 [(68.70 [(68.70 [(68.70 [(68.70 [(68.43), (62.97 [(6.97 [6.58) VS (80.48 [(68.48 [6.19)], (65.27 [5.45] VS (84.01 [2.76; WT%: (38.06 [4.15] VS (46.09 [46.09 [46.70)], (40.09 [(40.91 [(40.91 [(24.24(41.91+5.16) VS (64.64+3.86), P 0.05.
(3) Compared with 1 W after ADMSC treatment, the MPAP, RVHI, WA%, WA% and WT% of the rats in the treatment group were significantly higher [MPAP (mmHg): (23.07 [(23.07 [(mmHg): (23.07 [(23.07 [(22.98 [(22.98 [2.34) VS (18.63 [(2.34) VS (18.63 [2.15)]; (18.47 [(39.47 [(4.02 [(41.02 [0.93 [0.93] VS (36.21 [(36.21 [(21.21.21.27.27]], (62.97 [65.97 [65.27 [65.27: (40.91 (+ 5.24)), (41.91 (+ 5.16) VS (38.06 (+ 4.15)) There was no significant difference in MPAP, RVHI, WA% and WT% between ADMSC treated rats and ADMSC treated rats at 2 W and 3 W [MPAP (mmHg): (23.07 [2.84] VS (22.98 [2.34]; RVHI% (39.47 [4.02] VS (41.02 [0.93]; WA% (62.97 [6.58] VS (65.27 [5.45]); WT% (40.91 [5.24] VS (41.41] VS (5.05]].
(4) the colonization of GFP labeled ADMSC in lung was observed under fluorescence microscope.
CONCLUSION: ADMSC can effectively reduce pulmonary artery pressure, relieve right ventricular hypertrophy and improve pulmonary artery remodeling in monocrotaline-induced pulmonary hypertension rats; ADMSC only delays the progress of MCT-induced pulmonary hypertension after 1 W treatment, and the therapeutic effect of ADMSC after 2 W treatment reaches a stable level.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類(lèi)號(hào)】：R329

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