背景 结膜松弛症患者球结膜成纤维细胞中基质金属蛋白酶(MMPs)和组织金属蛋白酶抑制剂(TIMPs)的平衡失调,因此寻求一种药物以调节MMPs/TIMPs的平衡对于结膜松弛症的预防和治疗具有重要意义. 目的 研究杞精明目汤药物血清对体外培养的结膜松弛症患者球结膜成纤维细胞中MMPs和TIMPs表达的影响,探讨结膜松弛症的发病机制及杞精明目汤的作用. 方法 24只SD大鼠用杞精明目汤药物灌胃后3d收集腹主动脉血,制备含杞精明目汤药物的血清,以生理盐水灌胃的大鼠血清作为对照.收集接受结膜松弛症切除术患者的球结膜标本,用含质量分数10％胎牛血清的DMEM培养,分别将质量分数20％、15％、10％、5％的药物血清及8 ml/L表皮生长因子(EGF)加入细胞悬液中作用24 h,未加任何药物的结膜成纤维细胞组作为对照组.采用酶联免疫吸附(ELISA)法检测结膜成纤维细胞中MMP1、MMP3、TIMP1和TIMP3的表达(A值). 结果 培养的细胞呈长梭形,对波形蛋白表达呈阳性.不同质量分数药物血清作用后,人结膜成纤维细胞中MMP1的表达值明显下降,对照组、20％、15％、10％、5％药物血清组及EGF西药组结膜成纤维细胞中MMP1表达值的总体差异有统计学意义(F=466.664,P＜0.05),其中10％药物血清组MMP1的质量浓度为(9.92±0.14) mg/L,20％药物血清组为(11.87±0.11)mg/L,均明显低于对照组的(16.31±0.10) mg/L,差异均有统计学意义(t=99.974、87.394,P＜0.05)；不同质量浓度药物血清组、EGF西药组和对照组问结膜成纤维细胞中MMP3表达值的总体差异有统计学意义(F=158.168,P＜0.05),其中20％药物血清组MMP3质量浓度最低,为(3.50±0.03) mg/L,明显低于对照组的(4.44±0.11) mg/L,差异有统计学意义(t=21.991,P＜0.05).不同质量浓度药物血清组、对照组和EGF西药组间结膜成纤维细胞中TIMP1的表达值差异有统计学意义(F=183.508,P＜0.05),其中15％药物血清组TIMP1的表达量最低,为(1.88±0.06) mg/L,明显低于对照组的(3.20±0.32) mg/L,差异有统计学意义(t=10.353,P＜0.05)；不同质量浓度药物血清组、对照组和EGF西药组间结膜成纤维细胞中TIMP3的表达值差异有统计学意义(F=54.503,P＜0.05),其中20％药物血清组结膜成纤维细胞中TIMP3的表达量为(1.74±0.06) mg/L,高于对照组的(1.54±0.05) mg/L,差异有统计学意义(t=5.046,P=0.004),而15％、10％、5％药物血清组结膜成纤维细胞中TIMP3的表达量均低于对照组,差异均有统计学意义(P＜0.05). 结论 用含有20％杞精明目汤药物血清的培养基培养人眼结膜松弛症患者的球结膜成纤维细胞可下调细胞中MMP1、MMP3和TIMP1的表达量,并显著上调TIMP3的表达量,从而对结膜松弛症起到预防及治疗作用.
揹景 結膜鬆弛癥患者毬結膜成纖維細胞中基質金屬蛋白酶(MMPs)和組織金屬蛋白酶抑製劑(TIMPs)的平衡失調,因此尋求一種藥物以調節MMPs/TIMPs的平衡對于結膜鬆弛癥的預防和治療具有重要意義. 目的 研究杞精明目湯藥物血清對體外培養的結膜鬆弛癥患者毬結膜成纖維細胞中MMPs和TIMPs錶達的影響,探討結膜鬆弛癥的髮病機製及杞精明目湯的作用. 方法 24隻SD大鼠用杞精明目湯藥物灌胃後3d收集腹主動脈血,製備含杞精明目湯藥物的血清,以生理鹽水灌胃的大鼠血清作為對照.收集接受結膜鬆弛癥切除術患者的毬結膜標本,用含質量分數10％胎牛血清的DMEM培養,分彆將質量分數20％、15％、10％、5％的藥物血清及8 ml/L錶皮生長因子(EGF)加入細胞懸液中作用24 h,未加任何藥物的結膜成纖維細胞組作為對照組.採用酶聯免疫吸附(ELISA)法檢測結膜成纖維細胞中MMP1、MMP3、TIMP1和TIMP3的錶達(A值). 結果 培養的細胞呈長梭形,對波形蛋白錶達呈暘性.不同質量分數藥物血清作用後,人結膜成纖維細胞中MMP1的錶達值明顯下降,對照組、20％、15％、10％、5％藥物血清組及EGF西藥組結膜成纖維細胞中MMP1錶達值的總體差異有統計學意義(F=466.664,P＜0.05),其中10％藥物血清組MMP1的質量濃度為(9.92±0.14) mg/L,20％藥物血清組為(11.87±0.11)mg/L,均明顯低于對照組的(16.31±0.10) mg/L,差異均有統計學意義(t=99.974、87.394,P＜0.05)；不同質量濃度藥物血清組、EGF西藥組和對照組問結膜成纖維細胞中MMP3錶達值的總體差異有統計學意義(F=158.168,P＜0.05),其中20％藥物血清組MMP3質量濃度最低,為(3.50±0.03) mg/L,明顯低于對照組的(4.44±0.11) mg/L,差異有統計學意義(t=21.991,P＜0.05).不同質量濃度藥物血清組、對照組和EGF西藥組間結膜成纖維細胞中TIMP1的錶達值差異有統計學意義(F=183.508,P＜0.05),其中15％藥物血清組TIMP1的錶達量最低,為(1.88±0.06) mg/L,明顯低于對照組的(3.20±0.32) mg/L,差異有統計學意義(t=10.353,P＜0.05)；不同質量濃度藥物血清組、對照組和EGF西藥組間結膜成纖維細胞中TIMP3的錶達值差異有統計學意義(F=54.503,P＜0.05),其中20％藥物血清組結膜成纖維細胞中TIMP3的錶達量為(1.74±0.06) mg/L,高于對照組的(1.54±0.05) mg/L,差異有統計學意義(t=5.046,P=0.004),而15％、10％、5％藥物血清組結膜成纖維細胞中TIMP3的錶達量均低于對照組,差異均有統計學意義(P＜0.05). 結論 用含有20％杞精明目湯藥物血清的培養基培養人眼結膜鬆弛癥患者的毬結膜成纖維細胞可下調細胞中MMP1、MMP3和TIMP1的錶達量,併顯著上調TIMP3的錶達量,從而對結膜鬆弛癥起到預防及治療作用.
배경 결막송이증환자구결막성섬유세포중기질금속단백매(MMPs)화조직금속단백매억제제(TIMPs)적평형실조,인차심구일충약물이조절MMPs/TIMPs적평형대우결막송이증적예방화치료구유중요의의. 목적 연구기정명목탕약물혈청대체외배양적결막송이증환자구결막성섬유세포중MMPs화TIMPs표체적영향,탐토결막송이증적발병궤제급기정명목탕적작용. 방법 24지SD대서용기정명목탕약물관위후3d수집복주동맥혈,제비함기정명목탕약물적혈청,이생리염수관위적대서혈청작위대조.수집접수결막송이증절제술환자적구결막표본,용함질량분수10％태우혈청적DMEM배양,분별장질량분수20％、15％、10％、5％적약물혈청급8 ml/L표피생장인자(EGF)가입세포현액중작용24 h,미가임하약물적결막성섬유세포조작위대조조.채용매련면역흡부(ELISA)법검측결막성섬유세포중MMP1、MMP3、TIMP1화TIMP3적표체(A치). 결과 배양적세포정장사형,대파형단백표체정양성.불동질량분수약물혈청작용후,인결막성섬유세포중MMP1적표체치명현하강,대조조、20％、15％、10％、5％약물혈청조급EGF서약조결막성섬유세포중MMP1표체치적총체차이유통계학의의(F=466.664,P＜0.05),기중10％약물혈청조MMP1적질량농도위(9.92±0.14) mg/L,20％약물혈청조위(11.87±0.11)mg/L,균명현저우대조조적(16.31±0.10) mg/L,차이균유통계학의의(t=99.974、87.394,P＜0.05)；불동질량농도약물혈청조、EGF서약조화대조조문결막성섬유세포중MMP3표체치적총체차이유통계학의의(F=158.168,P＜0.05),기중20％약물혈청조MMP3질량농도최저,위(3.50±0.03) mg/L,명현저우대조조적(4.44±0.11) mg/L,차이유통계학의의(t=21.991,P＜0.05).불동질량농도약물혈청조、대조조화EGF서약조간결막성섬유세포중TIMP1적표체치차이유통계학의의(F=183.508,P＜0.05),기중15％약물혈청조TIMP1적표체량최저,위(1.88±0.06) mg/L,명현저우대조조적(3.20±0.32) mg/L,차이유통계학의의(t=10.353,P＜0.05)；불동질량농도약물혈청조、대조조화EGF서약조간결막성섬유세포중TIMP3적표체치차이유통계학의의(F=54.503,P＜0.05),기중20％약물혈청조결막성섬유세포중TIMP3적표체량위(1.74±0.06) mg/L,고우대조조적(1.54±0.05) mg/L,차이유통계학의의(t=5.046,P=0.004),이15％、10％、5％약물혈청조결막성섬유세포중TIMP3적표체량균저우대조조,차이균유통계학의의(P＜0.05). 결론 용함유20％기정명목탕약물혈청적배양기배양인안결막송이증환자적구결막성섬유세포가하조세포중MMP1、MMP3화TIMP1적표체량,병현저상조TIMP3적표체량,종이대결막송이증기도예방급치료작용.
Background Our previous study determined that expressions of matrix metalloproteinases (MMPs) and tissue matrix metalloproteinase inhibitors (TIMPs)change in the conjuntival fibroblasts of conjunctivochalasis in vitro.To seek a suitable drug is very important in the prevention and treatment of conjunctivochalasis.Objective This study was to explore the effect of qijingmingmu soup on the expressions of MMPs and TIMPs in human conjunctival fibroblasts of conjunctivochalasis.Methods Twenty-four SD rats were randomized into two groups.Qijingmingmu soup was administration gastrically for consecutive 3 days,and normal saline solution was given in the same way in the control group.The blood was collected from aortaventralis and drug serum was prepared.Human conjunctival samples were obtained during the surgery of conjunctivochalasis relaxation and cultured in the DMEM containing 10％ fetal bovine serum,20％,15％,10％,5％ of drug serum and 8 ml/L epidermal growth factor(EGF) was added into the medium respectively.Enzyme-linked immunosorbent assay(ELISA)was used to detect the expressions of MMP1,MMP3,TIMP1 and TIMP3in conjunctival fibroblasts.Results Cultured cells grew well with the fusiform shape and showed the positive response for vimentin.The expression value of MMP1 (A value)in the cells was declined after administration of qijingmingmu soup.A significant difference was found in the expression of MMP1 among the control group,20％,15％,10％,5％ drug serum groups and EGF group(F=466.664,P＜0.05),and that in the 10％ ([9.92±0.14] mg/L) and 20％ ([11.87 ±0.11] mg/L) drug serum groups was significantly lowed in comparison with the control group([16.31±0.10] mg/L)(t=99.974,87.394,P＜0.05).The expression value of the MMP3in the cells in the various drug serum groups,EGF group and the control group was significantly different(F=158.168,P＜0.05),with a lower value in the 20％ drug serum group compared with the control group ([3.50±0.03] mg/L vs.[4.44 ± 0.11] mg/L) (t =21.991,P ＜ 0.05).Also,the significantly different expressing value of TIMP1 was seen among all the groups (F=183.508,P＜0.05),and expressing value of TIMP1 in the 15％ drug serum group was(1.88±0.06)mg/L,which was lower than(3.20±0.32) mg/L of the control group(t=10.353,P＜0.05).Furthermore,the expressing value of the TIMP3 in the cells was significantly different among the various groups(F=54.503,P＜0.05),and that of the 20％ drug serum group was (1.743±0.065)mg/L and it was significantly higher than (1.54 ± 0.05) mg/L of the control group (t =5.046,P =0.004).However,the expressing value of TIMP3of the 15％,10％ and 5％ drug serum groups was lower than that of the control group,respectively all at(P＜0.05).Conclusions Qijingmingmu soup drug serum at the concentration of 20％ can down-regulate the expressions of MMP1,MMP3,TIMP1 and up-regulate the expression of TIMP3 in human conjunctivochalasis bulbar conjunctival fibroblastsin vitro,which probably plays preventive and therapeutic effects on conjunctivochalasis.
