延龄草甾体皂苷类成分对大鼠局灶性脑缺血/再灌注损伤的保护作用

Protective effect of steroidal saponins component of Trillium tschonoskii Maxim.on focal cerebral ischemia/reperfusion injury in rats

目的探讨延龄草中4个甾体皂苷类单体对大鼠局灶性脑缺血/再灌注(I/R)损伤的保护作用。方法将96只SD大鼠随机分为假手术组(Sham组),模型组(I/R组),尼莫地平片组(Contrast 1组),银杏叶片组(Contrast 2组),偏诺皂苷元-3-O-α-L-吡喃鼠李糖基(1→2)-β-D-吡喃葡萄糖苷(Y2T组)中、高剂量组,偏诺皂苷元-3-O-α-L-吡喃鼠李糖基(1→4)-[α-L-吡喃鼠李糖基(1→2)]-β-D-吡喃葡萄糖苷(简称Y3T组)中、高剂量组,偏诺皂苷元-3-O-α-L-吡喃鼠李糖基(1→4)-α-L-吡喃鼠李糖基(1→4)-[α-L-吡喃鼠李糖基(1→2)]-β-D-吡喃葡萄糖苷(Y4T组)中、高剂量组,偏诺皂苷元(YP组)中、高剂量组。采用改良Zea Longa线栓法构建局灶性脑I/R损伤动物模型,Sham组不插入线栓,其余操作同I/R组,各给药组在造模手术前连续给药8 d,造模后各组进行神经功能缺陷评分,然后麻醉处死,取材检测。干湿称重法检测大鼠脑指数,2,3,5-三苯基氯化四氮唑(TTC)染色法检测大鼠脑梗死体积,试剂盒法检测血清超氧化物歧化酶(SOD)、丙二醛(MDA)、谷氨酸的活性和一氧化氮(NO)、Ca^2+的含量,酶联免疫吸附试验检测大鼠脑组织中炎症介质肿瘤坏死因子(TNF)-α、白细胞介素(IL)-1β、IL-6的变化,苏木素-伊红(HE)染色观察海马CAI区和皮质层区病理学改变。结果与I/R组比较,给药各组能明显降低脑I/R大鼠翻正反射恢复时间、减轻I/R部位的病理损伤,降低脑组织含水量、减少脑梗死体积,提高SOD活性,降低MDA、谷氨酸、NO和Ca^2+含量,抑制激活小胶质细胞释放TNF-α、IL-1β、IL-6炎症因子,病理形态学有明显改善,差异均有统计学意义(P<0.05;P<0.01)。结论延龄草中4个甾体皂苷类单体对局灶性脑I/R损伤具有明显保护作用,其机制可能与抗炎及抗氧化应激有关。

Objective To investigate the anti-focal cerebral ischemia/reperfusion injury in rats active components of four steroidal saponins from Trillium tschonoskii Maxim.Methods Ninety-six male SD rats were randomly divided into twelve groups,the sham group,model group,Nimodiping group,Ginkgo leaf group,pennogenin-3-O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranoside high dose group and middle dose group,pennogenin-3-O-α-L-rhamnopyranosyl-(1→4)-[α-L-rhamnopyranosyl-(1→2)]-β-D-glucopyranoside high dose group and middle dose group,pennogenin-3-O-α-L-rhamnopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→4)-[α-L-rhamnopyranosyl-(1→2)]-β-D-glucopyranoside high dose group and middle dose group,pennogenin high dose group and middle dose group.Each group was ig administrated once a day for 8 days.The brain index and neurological function scores of twelve groups were observed,cerebral infarction area and various related enzymes(SOD,MDA,NO,glutamate,Ca^2+,TNF-α,IL-1β,IL-6)were determined.HE staining was used to evaluate the cerebral tissue morphology.Results Compared with model group,medication group could obviously decrease cerebral ischemia/reperfusion rats recovery time of righting reflex,alleviate the pathological damage of ischemia reperfusion location,reduce the water content of brain tissue,reduce the cerebral infarction volume,increase SOD activity and reduce MDA,glutamate,NO and Ca^2+ content,inhibit the activation of microglia releasing TNF-α,IL-1β and IL-6 inflammatory factors,improve pathological morpholog y,the differences were statistically significant(P<0.05 or P<0.01).Conclusions The four steroidal saponins of Trillium tschonoskii have obvious protective effect on focal cerebral ischemia/reperfusion injury,which might be related to anti-inflammatory and antioxidant stress.