摘要
目的基于血管内皮生长因子(VEGF)介导的信号通路研究丹蛭降糖胶囊对糖尿病大鼠血管新生的影响及作用机制。方法以雄性SD大鼠10只为空白组,另以50只GK大鼠建立下肢缺血模型,并随机分为模型组、丹蛭降糖胶囊高剂量组、丹蛭降糖胶囊中剂量组、丹蛭降糖胶囊低剂量组、盐酸吡格列酮组。分别以高中低剂量丹蛭降糖胶囊及盐酸吡格列酮灌胃30 d后,取血及下肢血管组织,镜下检测血管密度,酶联免疫吸附(ELISA)法检测血清VEGF、血管紧张素1、2(Ang-1、Ang-2)、血管生成素受体2(Tie-2)、一氧化氮(NO)、一氧化氮合酶(eNOS)相关指标的含量;蛋白质印迹法(Western blot)检测腓肠肌组织中Ang-1、Ang-2、Tie-2蛋白表达。结果糖尿病下肢缺血模型大鼠VEGF/Akt/eNOS信号通路的表达被抑制,治疗后丹蛭降糖胶囊高、中、低剂量组、盐酸吡格列酮组大鼠血清VEGF、eNOS、NO水平明显升高(P<0.01或P<0.05);腓肠肌组织中蛋白的表达也有类似的结果,丹蛭降糖胶囊高剂量组、盐酸吡格列酮组磷酸化蛋白激酶(p-Akt)、磷酸化一氧化氮合酶(p-eNOS)蛋白表达升高最为明显(P<0.01);高、中、低3个剂量组间比较,丹蛭降糖胶囊高剂量组p-Akt、p-eNOS蛋白表达明显升高(P<0.05)。另外,与模型组比较,各治疗组糖尿病大鼠血清及组织Ang-2水平明显降低(P<0.05或P<0.01),丹蛭降糖胶囊高剂量组降低更显著(P<0.01)。治疗后丹蛭降糖胶囊高、中、低剂量组、盐酸吡格列酮组大鼠血清及腓肠肌组织Ang-1、Tie-2表达均升高(P<0.05或P<0.01),丹蛭降糖胶囊高剂量组升高更显著(P<0.01)。结论丹蛭降糖胶囊能够激活糖尿病血管病变状态下被抑制的VEGF/Akt/eNOS及Ang/Tie2信号通路,促进血管基底膜的解离,为血管新生的启动做准备,促进下肢血管新生。
Objective To study the effect and mechanism of Danzhi Jiangtang Capsule(DZC) on angiogenesis based on VEGF-mediated signaling pathway in diabetic rats. Methods 10 SD rats were selected as the blank group, 50 GK rats were used to establish the model of lower limb ischemia in diabetes, and were randomly divided into the model group, the Danzhi Jiangtang Capsule high-dose group(H-DZC group), the Danzhi Jiangtang Capsule medium-dose group(M-DZC group), the Danzhi Jiangtang Capsule low-dose group(L-DZC group), and the pioglitazone hydrochloride group. In each group, the GK rats were gavaged correspondingly for 30 days.Then the density of new vessels in blood and vascular tissue of lower limbs was detected under the microscope;the contents of VEGF, Ang1, Ang2, Tie2, NO and NOS inserum were detected by ELISA;Ang1, Ang2 and Tie2 in gastrocnemius muscle were detected by Western Blot. Results The VEGF/Akt/eNOS signaling pathway was inhibited in diabetic rats with lower limb ischemia. After the treatment, serum levels of VEGF, eNOS and NO in the H-DZC, M-DZC and L-DZC groups and pioglitazone group were all significantly increased(P<0.01 or P<0.05). Similar results were also observed in the gastrocnemius muscle. The expression of p-Akt and P-eNOS in H-DZC group and pioglitazone hydrochloride group were most significantly increased(P<0.01).Among the three DZC groups, the expression of p-Akt and P-eNOS was most significantly increased in the high-dose group(P<0.05).In addition,compared with the model group, Ang-2 levels in serum and tissues in all treatment groups were significantly reduced(P<0.05 or P<0.01), and the decrease was more significant in H-DZC group(P<0.01). After the treatment, the expression of Ang-1 and Tie-2 in serum and gastrocnemius muscle tissues of rats in all treatment groups were increased(P<0.05 or P<0.01), and the increase was more significant in H-DZC group(P<0.01). Conclusion DZC seems to activate the inhibited VEGF/Akt/eNOS and Ang/Tie2 signaling pathways in diabetic vascular lesions and promote the dissociation of vascular basement membrane, so as to prepare for and promote the initiation of angiogenesis of vessels in lower limbs.
作者
倪英群
方朝晖
施慧
Ni Yingqun;Fang Zhaohui;Shi Hui(The First Affiliated Hospital of Anhui University of Chinese Medicine,Anhui 230038,China;Anhui University of Chinese Medicine,Anhui 230031,China)
出处
《北京中医药大学学报》
CAS
CSCD
北大核心
2020年第2期141-147,共7页
Journal of Beijing University of Traditional Chinese Medicine
基金
国家自然科学基金资助项目(No.81573944.No.81603602)
安徽省自然科学基金资助项目(No.1908085MH267)。
