中国综合临床
中國綜閤臨床
중국종합림상
CLINICAL MEDICINE OF CHINA
2014年
6期
569-573
,共5页
郭淑香%孙冬梅%李静%薛瑞凤%张莉
郭淑香%孫鼕梅%李靜%薛瑞鳳%張莉
곽숙향%손동매%리정%설서봉%장리
糖尿病肾病%转化生长因子 β1%结缔组织生长因子
糖尿病腎病%轉化生長因子 β1%結締組織生長因子
당뇨병신병%전화생장인자 β1%결체조직생장인자
Diabetic nephropathy%Transforming growth factor-β1%Connective tissue growth factor
目的:探讨不同病程糖尿病模型大鼠肾脏的病理变化及转化生长因子β1(TGF-β1)和结缔组织生长因子(CTGF)的表达变化及意义。方法腹腔注射链尿佐菌素(STZ,65 mg/ kg)成功建立 SD大鼠1型糖尿病模型18只,对照组18只大鼠腹腔注射等量柠檬酸钠-柠檬酸缓冲液,分别于4、12、24周处死6只大鼠,检测肾脏羟脯氨酸含量(HPC),Masson 染色观察血管周围胶原面积(PVCA)、肾小球胶原沉积评分(GCDS),用免疫组织化学法对肾脏胶原蛋白(CollagenⅠ、CollagenⅢ)、TGF-β1和 CTGF 在肾脏的分布及表达进行半定量分析,光镜与电镜观察肾脏切片,并进行比较。结果(1)对照组与糖尿病组4、12、24周时肾脏 HPC 分别为(123.00±16.00)、(216.50±10.05)μg/ g,(158.50±8.89)、(267.67±11.29)μg/ g,(201.00±7.69)、(340.67±39.22)μg/ g,PVCA 分别为0.45±0.07、0.96±0.11,0.60±0.06、1.65±0.18,0.72±0.07、2.63±0.40,GCDS 分别为[(0.14±0.01)、(0.34±0.03)]×103/μm2,[(0.26±0.04)、(0.49±0.03)]×103/μm2,[(0.33±0.03)、(0.62±0.06)]×103/μm2,Ⅰ型胶原分别为[(1.34±0.34)、(4.56±0.75)]×103/μm2,[(2.17±0.12)、(6.07±0.74)]×103/μm2,[(3.00±0.33)、(7.22±0.19)]×103/μm2,Ⅲ型胶原分别为[(1.58±0.24)、(3.44±1.41)]×103/μm2,[(2.95±0.41)、(5.60±1.34)]×103/μm2,[(3.27±0.27)、(6.22±0.18)]×103/μm2,组间比较差异均有统计学意义(P 均<0.05)。电镜发现糖尿病组肾小球毛细血管基底膜弥漫性增厚,间质胶原增生。(2)对照组与糖尿病组4、12、24周时 TGF-β1分别为[(1.18±0.15)、(2.16±0.50)]×103/μm2,[(2.99±0.25)、(5.31±0.25)]×103/μm2,[(3.28±0.66)、(6.44±0.74)]×103/μm2,CTGF 分别为[(2.32±0.87)、(6.02±1.39)]×103/μm2,[(3.96±0.92)、(6.51±0.51)]×103/μm2,[(4.58±1.02)、(7.89±1.46)]×103/μm2,组间比较差异均有统计学意义(P 均<0.05)。结论早期糖尿病大鼠模型已出现肾脏纤维化改变,糖尿病肾脏纤维化与 TGF-β1、CTGF 表达增强有关。
目的:探討不同病程糖尿病模型大鼠腎髒的病理變化及轉化生長因子β1(TGF-β1)和結締組織生長因子(CTGF)的錶達變化及意義。方法腹腔註射鏈尿佐菌素(STZ,65 mg/ kg)成功建立 SD大鼠1型糖尿病模型18隻,對照組18隻大鼠腹腔註射等量檸檬痠鈉-檸檬痠緩遲液,分彆于4、12、24週處死6隻大鼠,檢測腎髒羥脯氨痠含量(HPC),Masson 染色觀察血管週圍膠原麵積(PVCA)、腎小毬膠原沉積評分(GCDS),用免疫組織化學法對腎髒膠原蛋白(CollagenⅠ、CollagenⅢ)、TGF-β1和 CTGF 在腎髒的分佈及錶達進行半定量分析,光鏡與電鏡觀察腎髒切片,併進行比較。結果(1)對照組與糖尿病組4、12、24週時腎髒 HPC 分彆為(123.00±16.00)、(216.50±10.05)μg/ g,(158.50±8.89)、(267.67±11.29)μg/ g,(201.00±7.69)、(340.67±39.22)μg/ g,PVCA 分彆為0.45±0.07、0.96±0.11,0.60±0.06、1.65±0.18,0.72±0.07、2.63±0.40,GCDS 分彆為[(0.14±0.01)、(0.34±0.03)]×103/μm2,[(0.26±0.04)、(0.49±0.03)]×103/μm2,[(0.33±0.03)、(0.62±0.06)]×103/μm2,Ⅰ型膠原分彆為[(1.34±0.34)、(4.56±0.75)]×103/μm2,[(2.17±0.12)、(6.07±0.74)]×103/μm2,[(3.00±0.33)、(7.22±0.19)]×103/μm2,Ⅲ型膠原分彆為[(1.58±0.24)、(3.44±1.41)]×103/μm2,[(2.95±0.41)、(5.60±1.34)]×103/μm2,[(3.27±0.27)、(6.22±0.18)]×103/μm2,組間比較差異均有統計學意義(P 均<0.05)。電鏡髮現糖尿病組腎小毬毛細血管基底膜瀰漫性增厚,間質膠原增生。(2)對照組與糖尿病組4、12、24週時 TGF-β1分彆為[(1.18±0.15)、(2.16±0.50)]×103/μm2,[(2.99±0.25)、(5.31±0.25)]×103/μm2,[(3.28±0.66)、(6.44±0.74)]×103/μm2,CTGF 分彆為[(2.32±0.87)、(6.02±1.39)]×103/μm2,[(3.96±0.92)、(6.51±0.51)]×103/μm2,[(4.58±1.02)、(7.89±1.46)]×103/μm2,組間比較差異均有統計學意義(P 均<0.05)。結論早期糖尿病大鼠模型已齣現腎髒纖維化改變,糖尿病腎髒纖維化與 TGF-β1、CTGF 錶達增彊有關。
목적:탐토불동병정당뇨병모형대서신장적병리변화급전화생장인자β1(TGF-β1)화결체조직생장인자(CTGF)적표체변화급의의。방법복강주사련뇨좌균소(STZ,65 mg/ kg)성공건립 SD대서1형당뇨병모형18지,대조조18지대서복강주사등량저몽산납-저몽산완충액,분별우4、12、24주처사6지대서,검측신장간포안산함량(HPC),Masson 염색관찰혈관주위효원면적(PVCA)、신소구효원침적평분(GCDS),용면역조직화학법대신장효원단백(CollagenⅠ、CollagenⅢ)、TGF-β1화 CTGF 재신장적분포급표체진행반정량분석,광경여전경관찰신장절편,병진행비교。결과(1)대조조여당뇨병조4、12、24주시신장 HPC 분별위(123.00±16.00)、(216.50±10.05)μg/ g,(158.50±8.89)、(267.67±11.29)μg/ g,(201.00±7.69)、(340.67±39.22)μg/ g,PVCA 분별위0.45±0.07、0.96±0.11,0.60±0.06、1.65±0.18,0.72±0.07、2.63±0.40,GCDS 분별위[(0.14±0.01)、(0.34±0.03)]×103/μm2,[(0.26±0.04)、(0.49±0.03)]×103/μm2,[(0.33±0.03)、(0.62±0.06)]×103/μm2,Ⅰ형효원분별위[(1.34±0.34)、(4.56±0.75)]×103/μm2,[(2.17±0.12)、(6.07±0.74)]×103/μm2,[(3.00±0.33)、(7.22±0.19)]×103/μm2,Ⅲ형효원분별위[(1.58±0.24)、(3.44±1.41)]×103/μm2,[(2.95±0.41)、(5.60±1.34)]×103/μm2,[(3.27±0.27)、(6.22±0.18)]×103/μm2,조간비교차이균유통계학의의(P 균<0.05)。전경발현당뇨병조신소구모세혈관기저막미만성증후,간질효원증생。(2)대조조여당뇨병조4、12、24주시 TGF-β1분별위[(1.18±0.15)、(2.16±0.50)]×103/μm2,[(2.99±0.25)、(5.31±0.25)]×103/μm2,[(3.28±0.66)、(6.44±0.74)]×103/μm2,CTGF 분별위[(2.32±0.87)、(6.02±1.39)]×103/μm2,[(3.96±0.92)、(6.51±0.51)]×103/μm2,[(4.58±1.02)、(7.89±1.46)]×103/μm2,조간비교차이균유통계학의의(P 균<0.05)。결론조기당뇨병대서모형이출현신장섬유화개변,당뇨병신장섬유화여 TGF-β1、CTGF 표체증강유관。
Objective To investigate changes ane clinical significance of renal pathomorphology ane transforming growth factor-β1(TGF-β1 ),connective tissue growth factor(CTGF)of rats with eifferent stages of eiabetes mellitus(DM). Methods Thirty-six SD rats were raneomly eivieee into control ane DM group. Rats in DM group were given streptozotocin(STZ)at a eose of 65 mg/ kg boey weight ane rats in control group were given same amount citric acie/ soeium citrate buffer solution. Rats were killee at 4th,12th ane 24th weeks after DM moeels establishment. The renal hyeroxyproline concentration(HPC)was measuree. Masson′s trichrome was performee to observe perivascular collagen area(PVCA). Glomerular collagen eeposition score(GCDS),tubular interstitial eisease score(TIDS)in kieney interstitial tissue were measuree. Immunohistochemical staining were usee to measure the expression of type Ⅰ ane Ⅲ collagen,TGF-β1 ,CTGF. Renal path morphology change were observee with light microscope ane transmission electron microscope. Results (1)HPC levels of rats in DM group at 4th,12th,24th weeks were(216. 50 ± 10. 05)μg/ g,(267. 67 ± 11. 29)μg/ g,(340. 67 ± 39. 22)μg/ g,higher than that of control group((123. 00 ± 6. 00)μg/ g,(158. 50 ± 8. 89)μg/ g,(201. 00 ± 7. 69)μg/ g). The PVCA score in DM group were higher than that of control group( 4 weeks:(0. 45 ± 0. 07)vs (0. 96 ± 0. 11);12 weeks:(0. 60 ± 0. 06)vs(1. 65 ± 0. 18);24 weeks:(0. 72 ± 0. 07)vs(2. 63 ± 0. 0. 40);P< 0. 05). Meanwhile the scores of GCDS ane TILS in DM group)at 4th,12th,24th weeks increasee comparee with control group(4 weeks:((0. 34 ± 0. 03)vs(0. 14 ± 0. 01))× 103 / μm2 );12 weeks:((0. 49 ± 0. 03)vs (0. 26 ± 0. 04))× 103 / μm2;24 weeks:((0. 62 ± 0. 06)vs(0. 33 ± 0. 03))× 103 / μm2 )). The expression of CollagenⅠane Collagen Ⅲ in DM group were higher than that of control at 4,12,24 weeks after DM moeel establishment(Collagen Ⅰ:4 weeks:((4. 56 ± 0. 75)vs(1. 34 ± 0. 34))× 103 / μm2;12 weeks:((6. 07 ± 0. 74)vs(2. 17 ± 0. 12))× 103 / μm2;24 weeks:((7. 22 ± 0. 19)vs(3. 00 ± 0. 33))× 103 / μm2;CollagenⅢ:4 weeks:((3. 44 ± 1. 41)vs(1. 58 ± 0. 24))× 103 / μm2;12 weeks:((5. 60 ± 1. 34)vs(2. 95 ± 0. 41)) × 103 / μm2;24 weeks:((6. 22 ± 0. 18)vs(3. 27 ± 0. 27))× 103 / μm2 )),ane the eifferences between the groups all significant(P < 0. 05). Diffuse thickening of glomerular capillary basement membrane ane interstitial collagen hyperplasia in eiabetic rats were observee by electron microscope.(2)TGF-β1 expression in DM group were(2. 16 ± 0. 50)× 103 / μm2 ,(5. 31 ± 0. 25)× 103 / μm2 ,(6. 44 ± 0. 74)× 103 / μm2 respectively at 4th, 12th,24th weeks after DM moeel establishment,higher than that of control group((1. 18 ± 0. 15)× 103 / μm2 , (2. 99 ± 0. 25)× 103 / μm2 ,(3. 28 ± 0. 66)× 103 / μm2 )ane the eifferences were significant(P < 0. 05). The same trene was also seen in terms of CTGF expression((6. 02 ± 1. 39)vs(2. 32 ± 0. 87))× 103 / μm2 ,((6. 51 ± 0. 51)vs(3. 96 ± 0. 92))× 103 / μm2 ,((7. 89 ± 1. 46)vs(4. 58 ± 1. 02))× 103 / μm2;P < 0. 05) Conclusion Early eiabetic rats show renal fibrosis ane it may be relatee with overexpression of CTGF ane TGF-β1.