CAJ | 학술논문

松弛素/药理学%葡萄糖/投药和剂量%肾小球系膜细胞/代谢/药物作用%细胞外基质/药物作用%转化生长因子β1/代谢%肌动蛋白类/代谢%胶原Ⅳ型/代谢%纤连蛋白类/代谢鬆弛素/藥理學%葡萄糖/投藥和劑量%腎小毬繫膜細胞/代謝/藥物作用%細胞外基質/藥物作用%轉化生長因子β1/代謝%肌動蛋白類/代謝%膠原Ⅳ型/代謝%纖連蛋白類/代謝
송이소/약이학%포도당/투약화제량%신소구계막세포/대사/약물작용%세포외기질/약물작용%전화생장인자β1/대사%기동단백류/대사%효원Ⅳ형/대사%섬련단백류/대사
Relaxin/PD%Glucose/AD%Mesangial cells/ME/DE%Extracellular matrix/DE%Transforming growth factor beta1/ME%Actins/ME%Collagen type Ⅳ/ME%Fibronectins/ME

目的观察人松弛素(RLX)对高糖培养人肾小球系膜细胞分泌细胞外基质的影响,初步探讨其可能作用机制.方法将人肾小球系膜细胞株分组:正常糖组:含5.5 mmol/L葡萄糖;高糖组:含30 mmol/L葡萄糖;松弛素干预组:高糖(30 mmol/L葡萄糖)+松弛素(10 ng/ml);α-平滑肌动蛋白(α-SMA)的检测增加TGF-β1阳性对照组,即5.5 mmol/L葡萄糖+TGF-β1(5μg/L).CCK-8法检测松弛素对系膜细胞增殖的影响,ELISA法检测细胞上清中Ⅳ型胶原(ColⅣ)、纤连蛋白(FN)的表达.qPCR检测系膜细胞TGF-β1 mRNA表达.免疫印迹技术(Western blot)检测α-SMA表达,确定肾小球系膜细胞表型转化情况.结果松弛素对正常糖组及高糖组系膜细胞活力均未发现有影响;与正常糖组相比,高糖组各时间点肾系膜细胞分泌ColⅣ、FN显著增加,48 h尤为明显(57.28±0.59 vs 41.85 ±0.03、56.52±0.88 vs 33.80±0.24,P＜0.01),α-SMA表达增加(P＜0.05).与高糖组比较,松弛素干预组ColⅣ、FN表达在48 h下降最为明显(47.08±0.03 vs 57.28±0.59、36.16±0.52 vs 56.52±0.88,P＜0.01),α-SMA表达减少(P＜0.01),TGF-β1 mRNA表达下降(P＜0.01).结论松弛素能抑制高糖培养的系膜细胞分泌细胞外基质,部分作用机制是通过抑制细胞表型转分化及抑制TGF-β1基因表达,具有抗纤维化作用.
목적 관찰인송이소(RLX)대고당배양인신소구계막세포분비세포외기질적영향,초보탐토기가능작용궤제.방법 장인신소구계막세포주분조:정상당조:함5.5 mmol/L포도당;고당조:함30 mmol/L포도당;송이소간예조:고당(30 mmol/L포도당)+송이소(10 ng/ml);α-평활기동단백(α-SMA)적검측증가TGF-β1양성대조조,즉5.5 mmol/L포도당+TGF-β1(5μg/L).CCK-8법검측송이소대계막세포증식적영향,ELISA법검측세포상청중Ⅳ형효원(ColⅣ)、섬련단백(FN)적표체.qPCR검측계막세포TGF-β1 mRNA표체.면역인적기술(Western blot)검측α-SMA표체,학정신소구계막세포표형전화정황.결과 송이소대정상당조급고당조계막세포활력균미발현유영향;여정상당조상비,고당조각시간점신계막세포분비ColⅣ、FN현저증가,48 h우위명현(57.28±0.59 vs 41.85 ±0.03、56.52±0.88 vs 33.80±0.24,P＜0.01),α-SMA표체증가(P＜0.05).여고당조비교,송이소간예조ColⅣ、FN표체재48 h하강최위명현(47.08±0.03 vs 57.28±0.59、36.16±0.52 vs 56.52±0.88,P＜0.01),α-SMA표체감소(P＜0.01),TGF-β1 mRNA표체하강(P＜0.01).결론 송이소능억제고당배양적계막세포분비세포외기질,부분작용궤제시통과억제세포표형전분화급억제TGF-β1기인표체,구유항섬유화작용.
Objective To explore the effect and mechanism of relaxin on the production of extracellular matrix (ECM) excreted by high glucose stimulated human renal mesangial cells.Methods Cultured human mesangial cells (HMCs) were divided into three groups:(1) normal glucose group (NG,5.5 mmol/L D-glucose),(2) high glucose group (HG,30 mmol/L D-glucose),and (3) high glucose + relaxin group.Cell count kit (CCK8) was used to examine the cell proliferation.The levels of fibronectin and collagen type Ⅳ in the culture supernatants were examined with a solid-phase enzyme-linked immunoadsorbent assay (ELISA);Western blot method was used to detect the expression of α-smooth muscle actin (α-SMA) protein.The transforming growth factor-β1 (TGF-β1) mRNA expression was detected with quantitative polymerase chain reaction (qPCR) method.Results No proliferation and inhibition effects were observed in both normal and high glucose group.Compared to the normal glucose group,the levels of fibronectin,and collagen type Ⅳ increased significantly (57.28 ± 0.59 vs 41.85 ± 0.03,56.52 ± 0.88 vs 33.80 ± 0.24,P ＜ 0.01)after cultured 48 h in high concentration of glucose.Compared to the high glucose group,a significantly decreases of fibronectin and collagen type Ⅳ (47.08 ± 0.03 vs 57.28 ± 0.59,36.16 ± 0.52 vs 56.52 ±0.88,P ＜0.01) were observed in the relaxin treated group.The expressions of α-smooth muscle actin and TGF-β1 were decreased (P ＜0.01).Conclusions Relaxin can suppress the overproduction of ECM excreted by HMC cultured in high ambient glucose,and its mechanism is partly due to the inhibition of TGF-β1.