首页|蠲痹通络口服液改善2型糖尿病模型小鼠坐骨神经细胞凋亡的效用评价研究

蠲痹通络口服液改善2型糖尿病模型小鼠坐骨神经细胞凋亡的效用评价研究

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目的 评价蠲痹通络口服液对 2 型糖尿病模型小鼠坐骨神经细胞凋亡的改善效应.方法 小鼠给予高脂高糖饲料联合链脲霉素(STZ)腹腔注射构建 2 型糖尿病小鼠模型,分别给予阳性药二甲双胍(200 mg·kg-1·d-1)、蠲痹通络口服液低剂量(3.9 g·kg-1·d-1)与高剂量(7.8 g·kg-1·d-1),连续给药 35 d.采用热板法检测动物疼痛潜伏期;测定受试动物空腹血糖值、胰岛素值、糖化血红蛋白值;采用生化试剂盒检测小鼠血清总胆固醇(T-CHO)、三酰甘油(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)与氧化应激标志物超氧化物歧化酶(SOD)、过氧化氢酶(CAT)与谷胱甘肽过氧化物酶(GSH-Px)活力以及氧化产物丙二醛(MDA)表达;采用ELISA法检测小鼠血清中肿瘤坏死因子α(TNF-α)、白介素(IL)-1β、IL-6 与IL-10 水平;采用HE染色考察模型小鼠坐骨神经损伤情况;采用TUNEL法检测小鼠坐骨神经凋亡情况;采用Western blot法检测小鼠坐骨神经凋亡信号通路蛋白cleaved Caspase-3、Caspase-3,神经丝蛋白NF-L、NF-H与氧化应激信号通路蛋白Nrf2、HO-1 表达.结果 高剂量蠲痹通络口服液缩短模型小鼠热痛反应潜伏期(P<0.01);下调模型小鼠空腹血糖与糖化血红蛋白(P<0.01),上调空腹血浆胰岛素(P<0.01);下调血清T-CHO、TG、LDL-C水平(P<0.01),上调HDL-C水平(P<0.01);下调血清中促炎因子TNF-α、IL-1β与IL-6 水平(P<0.05),上调抗炎细胞因子IL-10 水平(P<0.01);抑制坐骨神经结构破坏与凋亡(P<0.05);下调凋亡通路蛋白cleaved Caspase-3 与Caspase-3 的比值(P<0.01);上调坐骨神经组织神经丝蛋白NF-L与NF-H表达(P<0.05,P<0.01);上调抗氧化应激蛋白Nrf2 与HO-1 的表达(P<0.01).结论 蠲痹通络口服液具有改善 2 型糖尿病模型小鼠坐骨神经细胞凋亡的作用,可能与其改善氧化应激与炎性应激作用相关.
Evaluation of Juanbi Tongluo Oral Liquid on Improving Sciatic Neuronal Apoptosis in Type 2 Diabetic Model Mice
OBJECTIVE To evaluate the ameliorative effect of Juanbi Tongluo Oral Liquid on sciatic neuronal apoptosis in Type 2 diabetic model mice.METHODS The Type 2 diabetes mouse model was established by feeding with high-fat and high-sugar diet combined with intraperitoneal injection of streptozotocin(STZ).The mice were treated with metformin(200 mg·kg-1·d-1),low dose(3.9 g·kg-1·d-1)and high dose(7.8 g·kg-1·d-1)Juanbi Tongluo Oral Liquid for 35 days.The latency of response to thermal stimu-lation was detected by hot plate,and the values of blood glucose insulin and glycosylated hemoglobin were determined.Biochemical kits were used to detect the expression of serum total cholesterol(T-CHO),triglyceride(TG),high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol(LDL-C),superoxide dismutase(SOD),catalase(CAT),glutathione peroxidase(GSH-Px)and oxidation product malondialdehyde(MDA).The expression of tumor cytokine α(TNF-α),interleukin(IL)-1β,IL-6 and IL-10 in serum of mice were detected by ELISA method;the injury of sciatic nerve of model mice was detected by HE staining;the apoptosis of sciatic nerve was detected by TUNEL method;and the expression of neurofilament protein NF-L,apoptosis(cleaved Caspase-3,Caspase-3)and oxidative stress(Nrf2,HO-1)signal pathway proteins in sciatic nerve of model mice were detected by Western blot method.RESULTS High-dose Juanbi Tongluo Oral Liquid shortened the latent period of heat pain response in model mice(P<0.01);downregulated fasting blood glucose and glycated hemoglobin(P<0.01),and upregulated fasting plasma insulin in model mice(P<0.01);downregulated serum T-CHO,TG,and LDL-C levels(P<0.01),upregulated HDL-C levels(P<0.01);downregulated serum pro-inflammatory factors TNF-α,IL-1β and IL-6 levels(P<0.05),upregulated the anti-inflammatory cyto-kine IL-10 level(P<0.01);inhibited sciatic nerve structural damage and apoptosis(P<0.05);downregulated the ratio of cleaved Caspase-3 to Caspase-3 in the apoptosis pathway(P<0.01);upregulated the expression of neurofilament proteins NF-L and NF-H in sciatic nerve tissue(P<0.05,P<0.01);and upregulated the expression of antioxidant stress proteins Nrf2 and HO-1(P<0.01).CONCLUSION Juanbi Tongluo Oral Liquid can improve sciatic neuronal apoptosis of Type 2 diabetic mice,which may be related to its effect on improving oxidative stress and inflammatory stress.

Juanbi Tongluo Oral Liquiddiabetic peripheral neuropathysciatic nerveapoptosis

黄菲、仝佳祥、刘伟、刘梦秋、陈婷、刘元君、张明英、吴雨辰、魏微、赵雨婷、田列、朱悦

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南京中医药大学附属苏州市中医医院,江苏 苏州 215007

南京中医药大学药学院,江苏省方剂研究重点实验室/江苏省方剂高技术研究重点实验室/中药资源产业化与方剂创新药物国家地方联合工程研究中心/江苏省中药资源产业化过程协同创新中心,江苏 南京 210023

蠲痹通络口服液 糖尿病周围神经病变 坐骨神经 凋亡

江苏省干部保健科研项目苏州市民生科技示范工程项目姑苏卫生人才培训项目南京中医药大学自然科学基金重点项目南京中医药大学自然科学基金重点项目&&苏州市卫生健康委员会青年科技项目

BJ19009SS202006GSWS2020079XZR2023021XZR2021043SKYD2023041KJXW2023049

2024

10.14148/j.issn.1672-0482.2024.0932

南京中医药大学学报
南京中医药大学

南京中医药大学学报

CSTPCD北大核心
影响因子：1.658
ISSN：1672-0482
年,卷(期)：2024.40(9)
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