Sha Qiankun, Huang Yan, Zhou Faying, Wang Yong, Fang Wei, Li Jing
Department of Pharmacy, Chongqing Yangdu Biology Institute Chongqing 408000, China.
Department of Pharmacy, The People's Hospital of Nanchuan Chongqing 408499, China.
Am J Transl Res. 2023 Feb 15;15(2):995-1006. eCollection 2023.
To analyze the effect of polydatin (PD) from Huhuang Shaoshang Liniment on oxidative damage and inflammatory response in rats with cerebral ischemia-reperfusion (CI/R), and the effect of PD on NF-E2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1)/NAD(P)H quinone oxidoreductase 1 (NQO1) pathway.
The rat model of focal CI/R was established using suture-occlusion method and treated with intraperitoneal injection of PD (25 mg/kg and 50 mg/kg). High performance liquid chromatography (HPLC) was used to determine the PD content in three batches of Huhuang Shaoshang Liniment. The neurological function scores of each group were observed at 24 h, 48 h, and 72 h postoperatively. The hematoxylin and eosin (HE) staining was conducted to observe the morphological structure of brain tissue, the triphenyltetrazolium chloride (TTC) staining was used to determine the size of cerebral infarction, and the neuronal apoptotic index was calculated using image analysis system under the optical microscope. The expressions of the Nrf2/HO-1/NQO1 pathway and neuronal apoptosis-related proteins in brain tissue were measured using Western blot.
The PD content in three batches of Huhuang Shaoshang Liniment was detected by HPLC, and the average result showed that the product contained 0.73 mg PD per 1 mL. The PD 50 mg/kg group and 25 mg/kg group showed lower neurological function scores at 24 h, 48 h, and 72 h postoperatively, lower percentage of cerebral infarction area on the ischemic side and apoptotic index, lower interleukin (IL)-1β, IL-6, malondialdehyde (MDA), and lactate dehydrogenase (LDH) levels, and lower Bax protein expression (P < 0.05), and showed higher IL-4, IL-10, superoxide dismutase (SOD), glutathione peroxidase (GPx), Nrf2, HO-1, NQO1, and c-Myc protein expression than the CI/R group (P < 0.05).
PD from Huhuang Shaoshang Liniment can alleviate neurological damage, improve neuronal morphology and structure, reduce cerebral infarction area, alleviate oxidative damage and inflammatory response, and inhibit neuronal cell apoptosis in CI/R model rats.
分析虎黄烧伤酊中虎杖苷（PD）对脑缺血再灌注（CI/R）大鼠氧化损伤和炎症反应的影响，以及PD对核因子E2相关因子2（Nrf2）/血红素加氧酶-1（HO-1）/烟酰胺腺嘌呤二核苷酸磷酸（NAD(P)H）醌氧化还原酶1（NQO1）通路的影响。
采用线栓法建立局灶性CI/R大鼠模型，腹腔注射PD（25mg/kg和50mg/kg）进行治疗。采用高效液相色谱法（HPLC）测定三批虎黄烧伤酊中PD的含量。术后24h、48h和72h观察各组神经功能评分。进行苏木精-伊红（HE）染色观察脑组织形态结构，采用氯化三苯基四氮唑（TTC）染色测定脑梗死面积，在光学显微镜下利用图像分析系统计算神经元凋亡指数。采用蛋白质免疫印迹法检测脑组织中Nrf2/HO-1/NQO1通路及神经元凋亡相关蛋白的表达。
通过HPLC检测三批虎黄烧伤酊中PD的含量，平均结果显示该产品每1mL含0.73mg PD。PD 50mg/kg组和25mg/kg组术后24h、48h和72h神经功能评分较低，缺血侧脑梗死面积百分比和凋亡指数较低，白细胞介素（IL）-1β、IL-6、丙二醛（MDA）和乳酸脱氢酶（LDH）水平较低，Bax蛋白表达较低（P<0.05），且与CI/R组相比，IL-4、IL-10、超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GPx）、Nrf2、HO-1、NQO1和c-Myc蛋白表达较高（P<0.05）。
虎黄烧伤酊中的PD可减轻CI/R模型大鼠的神经损伤，改善神经元形态结构，缩小脑梗死面积，减轻氧化损伤和炎症反应，抑制神经元细胞凋亡。
