止血明目方对糖尿病视网膜病变大鼠炎症因子和氧化应激的影响

Effect of Zhixue Mingmu Formula on inflammatory factors and oxidative stress in diabetic retinopathy rats

目的观察止血明目方(ZXMM)对糖尿病视网膜病变(DR)大鼠炎症因子和氧化应激的影响,探讨其作用机制。方法SD大鼠经单次腹腔注射链脲佐菌素诱导DR模型,随机分为模型组(MG)、阳性对照药羟苯磺酸钙组(CD)、ZXMM低剂量组(LZXMM)和高剂量组(HZXMM),并以正常大鼠作为对照组(CG),每组6只。CD给予羟苯磺酸钙胶囊水溶剂135 mg/kg,LZXMM、HZXMM分别按照30.6、45.9 g/kg浓度给予ZXMM药液,CG和MG给予等体积生理盐水,每日1次,连续灌胃30 d。测量各组大鼠体重、血糖值,采用苏木精-伊红染色法观察大鼠视网膜病理改变情况,酶联免疫吸附法检测大鼠血清及视网膜组织中白细胞介素-6(IL-6)、可溶性细胞间黏附分子1(sICAM-1)含量、血管内皮生长因子A(VEGF-A)水平和超氧化物歧化酶(SOD)活性。结果(1)体重和血糖:与CG比较,MG、CD、LZXMM和HZXMM大鼠体重减轻(t_(MG)=4.621,P=0.001;t_(CD)=7.465,P=0.000;t_(LZXMM)=5.338,P=0.000;t_(HZXMM)=7.712,P=0.000),血糖升高(t_(MG)=-52.809,t_(CD)=-34.659,t_(LZXMM)=-39.717,t_(HZXMM)=-49.120,均P=0.000),差异均有统计学意义。(2)视网膜形态:与CG比较,MG大鼠视网膜组织病理损伤明显;与MG比较,CD、LZXMM和HZXMM大鼠视网膜病理损伤减轻。(3)血清和视网膜中相关因子,①IL-6:与CG比较,MG大鼠血清和视网膜IL-6水平升高(血清:t=-8.090,视网膜:t=-8.946,均P=0.000);与MG比较,CD、LZXMM和HZXMM大鼠血清和视网膜IL-6水平降低(血清:t_(CD)=6.445,t_(LZXMM)=5.237,t_(HZXMM)=7.190,均P=0.000。视网膜:t_(CD)=6.067,P=0.000;t_(LZXMM)=3.907,P=0.003;t_(HZXMM)=8.004,P=0.000),差异均有统计学意义。②sICAM-1:与CG比较,MG大鼠血清和视网膜sICAM-1水平升高(血清:t=-13.221,视网膜:t=-18.487,均P=0.000);与MG比较,CD、LZXMM和HZXMM大鼠血清和视网膜sICAM-1水平降低(血清:t_(CD)=6.584,P=0.000;t_(LZXMM)=4.301,P=0.002,t_(HZXMM)=5.891,P=0.000。视网膜:t_(CD)=7.522,t_(LZXMM)=12.737,t_(HZXMM)=14.507,均P=0.000),差异均有统计学意义。③VEGFA:与CG比较,MG大鼠血清和视网膜VEGF-A水平升高(血清:t=-7.135,视网膜:t=-20.695,均P=0.000);与MG比较,CD和HZXMM大鼠血清和视网膜、LZXMM大鼠视网膜VEGF-A水平降低(血清:t_(CD)=6.734,P=0.000;t_(HZXMM)=4.243,P=0.002。视网膜:t_(CD)=7.208,t_(LZXMM)=5.620,t_(HZXMM)=13.648,均P=0.000),差异均有统计学意义。④SOD:与CG比较,MG大鼠血清和视网膜SOD水平降低(血清:t=7.442,视网膜:t=10.718,均P=0.000);与MG比较,CD、LZXMM和HZXMM大鼠血清和视网膜SOD水平升高(血清:t_(CD)=-3.193,P=0.010;t_(LZXMM)=-2.346,P=0.041;t_(HZMXX)=-5.021,P=0.001。视网膜:t_(CD)=-7.919,t_(LZXMM)=-8.314,t_(HZMXX)=-10.054,均P=0.000),差异均有统计学意义。结论ZXMM能改善DR大鼠视网膜病理损伤,其机制可能与抑制机体及视网膜组织炎症因子并增强其抗氧化应激能力有关。

OBJECTIVE To investigate the effects of Zhixue Mingmu Formula(ZXMM)on inflammation factors and oxidative stress in diabetic retinopathy(DR)rats and to explore the pharmacological mechanism of this formula.METHODS The SD rats were injected streptozotocin intraperitoneally to induce DR.Then DR rats were randomly divided into model group,calcium dobesilate(CD)group,low-dose ZXMM(LZXMM)group and high-dose ZXMM(HZXMM)group,with normal rats as control group.Each group comprised of 6 rats.Rats in CD group were given 135 mg/kg of calcium dobesilate by gavage.Rats in LZXMM and HZXMM group were given 30.6 g/kg and 45.9 g/kg of ZXMM respectively.Rats in control and model group were given normal saline of equal volume once a day,continuously for thirty days.Then body weight and blood glucose levels of rats were measured;Pathological changes of the retina were observed by HE staining;IL-6,sICAM-1 and VEGF-A levels and SOD activity in serum and retinal tissues were detected using ELISA kit.RESULTS(1)Body weight and blood glucose:Compared with rats in control group,counterparts in model,CD,LZXMM and HZXMM group showed significant body weight loss(t_(MG)=4.621,P=0.001;t_(CD)=7.465,P=0.000;t_(LZXMM)=5.338,P=0.000;t_(HZXMM)=7.712,P=0.000)and body glucose rise(t_(MG)=-52.809,t_(CD)=-34.659,t_(LZXMM)=-39.717,t_(HZXMM)=-49.120,all P=0.000),the differences were all statistically significant.(2)Retinal morphology:Compared with rats in control group,the retinal pathological damage was more obvious in model group;Compared with rats in model group,the pathological damage of retina was alleviated in CD group,LZXMM and HZXMM group.(3)Related factors in serum and retina,①IL-6:Compared with control group,serum and retinal IL-6 levels of model group rats increased(serum:t=-8.090,retina:t=-8.946,all P=0.000);Compared with model group,serum and retinal IL-6 levels of CD,LZXMM and HZXMM group rats decreased(Serum:t_(CD)=6.445,t_(LZXMM)=5.237,t_(HZXMM)=7.190,all P=0.000.Retinal:t_(CD)=6.067,P=0.000;t_(LZXMM)=3.907,P=0.003;t_(HZXMM)=8.004,P=0.000),the differences were statistically significant.②sICAM-1:Compared with control group,the levels of sICAM-1 in serum and retina of model group rats increased(serum:t=-13.221,retina:t=-18.487,all P=0.000);Compared with model group,rat serum and retinal sICAM-1 levels of CD,LZXMM and HZXMM group decreased(Serum:t_(CD)=6.584,P=0.000;t_(LZXMM)=4.301,P=0.002,t_(HZXMM)=5.891,P=0.000.Retina:t_(CD)=7.522,t_(LZXMM)=12.737,t_(HZXMM)=14.507,all P=0.000),the differences were statistically significant.③VEGF-A:Compared with control group,the levels of VEGF-A in serum and retina of model rats increased(serum:t=-7.135,retina:t=-20.695,all P=0.000);Compared with model group,retinal VEGF-A levels of CD and HZXMM group rats serum and retina,LZXMM group rats retina decreased(Serum:t_(CD)=6.734,P=0.000;t_(HZXMM)=4.243,P=0.002.Retinal:t_(CD)=7.208,t_(LZXMM)=5.620,t_(HZXMM)=13.648,all P=0.000),the differences were statistically significant.④SOD:Compared with control group the serum and retinal SOD levels of model group rats decreased(serum:t=7.442,retina:t=10.718,all P=0.000);Compared with model group,the serum and retinal SOD levels of CD,LZXMM and HZXMM group rats increased(Serum:t_(CD)=-3.193,P=0.010;t_(LZXMM)=-2.346,P=0.041;t_(HZMXX)=-5.021,P=0.001.Retina:t_(CD)=-7.919,t_(LZXMM)=-8.314,t_(HZMXX)=-10.054,all P=0.000),the differences were statistically significant.CONCLUSIONS The ZXMM alleviates pathological damage of the retina in DR rats which is related to the inhibition of inflammatory factors and the enhancement of antioxidant stress ability.