摘要
目的基于Piezo1/ROS/SERCA1a信号轴探讨针刺抑制膝骨关节炎(KOA)模型大鼠股四头肌肌细胞凋亡的机制。方法选择SPF级Wistar大鼠36只,采用随机数字表法分为假手术组、模型组、塞来昔布组与针刺组,每组9只。模型组、塞来昔布组与针刺组采用改良Hulth法造模,假手术组仅切开关节腔后缝合。造模后假手术组、模型组给予5 mL生理盐水灌胃,塞来昔布组给予5 mL浓度为1.8 mg/100 g的塞来昔布溶液灌胃,针刺组给予针刺髌下、鹤顶次、血海次。每日干预1次,连续干预14 d。测量各组大腿围度、股四头肌湿重并计算湿重维持率及湿重体质量比值;采用HE染色法观察各组股四头肌形态;采用TUNEL法检测各组股四头肌肌细胞凋亡率;采用免疫荧光染色法检测股四头肌肌细胞内活性氧(ROS)表达水平以及Piezo1与SERCA1a共表达情况;采用Western blot法检测各组Piezo1、SERCA1a及凋亡相关蛋白Bcl-2、BCL-XL、Bax,Cyt C,Caspase-3蛋白相对表达量。结果与假手术组比较,模型组大腿围度较小(P<0.05),股四头肌湿重较轻(P<0.05),湿重维持率及湿重体质量比值均较低(P<0.05);与模型组比较,塞来昔布组和针刺组大腿围度均较大(P<0.05),股四头肌湿重均较重(P<0.05),湿重维持率及湿重体质量比值均较高(P<0.05);与塞来昔布组比较,针刺组大腿围度较大(P<0.05),股四头肌湿重较重(P<0.05),湿重维持率及湿重体质量比值均较高(P<0.05)。与假手术组比较,模型组股四头肌肌细胞凋亡率较高(P<0.05);与模型组比较,塞来昔布组和针刺组股四头肌肌细胞凋亡率均较低(P<0.05);与塞来昔布组比较,针刺组股四头肌肌细胞凋亡率较低(P<0.05)。与假手术组比较,模型组股四头肌肌细胞内ROS表达水平较高(P<0.05);与模型组比较,塞来昔布组和针刺组股四头肌肌细胞内ROS表达水平均较低(P<0.05);与塞来昔布组比较,针刺组股四头肌肌细胞内ROS表达水平较低(P<0.05)。与假手术组比较,模型组Piezo1、SERCA1a蛋白表达水平较低(P<0.05);与模型组比较,塞来昔布组和针刺组Piezo1、SERCA1a蛋白表达水平均较高(P<0.05);与塞来昔布组比较,针刺组Piezo1、SERCA1a蛋白表达水平均较高(P<0.05)。与假手术组比较,模型组Piezo1、SERCA1a蛋白相对表达量较低(P<0.05);与模型组比较,塞来昔布组和针刺组Piezo1、SERCA1a蛋白相对表达量均较高(P<0.05);与塞来昔布组比较,针刺组Piezo1、SERCA1a蛋白相对表达量较高(P<0.05)。与假手术组比较,模型组BCL-XL蛋白相对表达量较低(P<0.05),Cyt C、Caspase-3蛋白相对表达量较高(P<0.05),Bax/Bcl-2比值较高(P<0.05);与模型组比较,塞来昔布组和针刺组BCLXL蛋白相对表达量均较高(P<0.05),Cyt C、Caspase-3蛋白相对表达量均较低(P<0.05),Bax/Bcl-2比值均较低(P<0.05);与塞来昔布组比较,针刺组BCL-XL蛋白相对表达量较高(P<0.05),Cyt C、Caspase-3蛋白相对表达量较低(P<0.05),Bax/Bcl-2比值较低(P<0.05)。结论针刺可抑制KOA模型大鼠股四头肌肌细胞凋亡,其机制可能与调节Piezo1/ROS/SERCA1a信号轴有关。
Objective To explore the mechanism by which acupuncture inhibits quadriceps femoris myocyte apoptosis in a rat model of knee osteoarthritis(KOA)based on the Piezo1/ROS/SERCA1a signaling axis.Methods A total of thirty-six SPF-grade Wistar rats were randomly divided into sham-operated group,model group,celecoxib group,and acupuncture group,with nine rats in each group.The KOA model was established in the model,celecoxib,and acupuncture groups using the modified Hulth method,while the sham-operated group only underwent arthrotomy followed by suture.After modeling,the sham-operated and model groups received 5 mL of normal saline by gavage,the celecoxib group received 1.8 mg/mL celecoxib solution 5 mL by gavage,and the acupuncture group received acupuncture at the infra-patellar,Hedingci,and Xuehaici acpoints.Interventions were performed once daily for 14 consecutive days.Thigh circumference and quadriceps wet weight were measured,and the wet weight maintenance rate and wet weight to body weight ratio were calculated.HE staining was used to observe quadriceps morphology;TUNEL assay was applied to detect quadriceps myocyte apoptosis rate;immunofluorescence staining was employed to assess intracellular ROS expression and co‑expression of Piezo1 and SERCA1a in quadriceps myocytes;and Western blot was performed to measure the relative expression levels of Piezo1,SERCA1a,and apoptosis-related proteins(Bcl-2,BCL-XL,Bax,Cyt C,Caspase-3).Results Compared with the sham-operated group,the model group showed a smaller thigh circumference(P<0.05),lighter wet weight of the quadriceps femoris(P<0.05),and lower wet weight maintenance rate and wet weight to body weight ratio(P<0.05).Compared with the model group,the thigh circumference was greater(P<0.05),quadriceps wet weight was heavier(P<0.05);and both the wet weight maintenance rate and the wet weight to body weight ratio were higher in the celecoxib and acupuncture groups(P<0.05).Compared with the celecoxib group,the thigh circumference was larger(P<0.05),quadriceps wet weight was heavier(P<0.05),and both the wet weight maintenance rate and the wet weight to body weight ratio were higher in the acupuncture group(P<0.05).Compared with the sham-operated group,the apoptosis rate of quadriceps myocytes was higher in the model group(P<0.05);compared with the model group,the apoptosis rate of quadriceps myocytes was lower in both the celecoxib group and the acupuncture group(P<0.05);and compared with the celecoxib group,the apoptosis rate of quadriceps myocytes was lower in acupuncture group(P<0.05).Compared with the sham-operated group,intracellular ROS expression level in quadriceps myocytes was higher in the model group(P<0.05);compared with the model group,intracellular ROS expression levels in quadriceps myocytes were lower in the celecoxib group and the acupuncture group(P<0.05);and compared with the celecoxib group,intracellular ROS expression level in quadriceps myocytes was lower in the acupuncture group(P<0.05).Compared with the sham-operated group,the protein expression levels of Piezo1 and SERCA1a were lower in the model group(P<0.05);compared with the model group,the protein expression levels of Piezo1 and SERCA1a were higher in the celecoxib group and the acupuncture group(P<0.05);and compared with the celecoxib group,the protein expression levels of Piezo1 and SERCA1a were higher in the acupuncture group(P<0.05).Compared with the sham-operated group,the relative expression levels of Piezo1 and SERCA1a proteins in the model group were lower(P<0.05);compared with the model group,the relative expression levels of Piezo1 and SERCA1a proteins were higher in the celecoxib group and the acupuncture group(P<0.05);compared with the celecoxib group,the relative expression levels of Piezo1 and SERCA1a proteins were higher in the acupuncture group(P<0.05).Compared with the sham-operated group,the relative expression level of BCL-XL protein in the model group was lower(P<0.05),the relative expression levels of Cyt C and Caspase-3 proteins were higher(P<0.05),and the Bax/Bcl-2 ratio was higher(P<0.05).Compared with the model group,the relative expression of BCL-XL protein was higher,the relative expression of Cyt C and Caspase3 proteins were lower,and the Bax/Bcl-2 ratio was lower in the celecoxib group and acupuncture group(P<0.05).Compared with the celecoxib group,the relative expression of BCL-XL protein was higher,Cyt C and Caspase-3 proteins were lower,and the Bax/Bcl-2 ratio was lower in the acupuncture group(P<0.05).Conclusion Acupuncture can inhibit quadriceps myocyte apoptosis in KOA model rats,and its mechanism may be related to the regulation of the Piezo1/ROS/SERCA1a signaling axis.
作者
郑曲
董宝强
林星星
张宇
关雪峰
韩易言
ZHENG Qu;DONG Baoqiang;LIN Xingxing;ZHANG Yu;GUAN Xuefeng;HAN Yiyan(Liaoning University of Traditional Chinese Medicine,Shenyang,Liaoning 110847,China;Liaoning Provincial Key Laboratory of Acupuncture Biology,Shenyang,Liaoning 110847,China;Liaoning Provincial Key Laboratory of Acupuncture and Moxibustion Health and Rehabilitation,Shenyang,Liaoning 110847,China;Shenyang Pharmaceutical University,Shenyang,Liaoning 110016,China)
出处
《康复学报》
2026年第1期56-65,共10页
Rehabilitation Medicine
基金
国家重点研发计划项目(2021YFC1712800)
辽宁省科技计划联合计划(自然科学基金-面上项目)(2025-MSLH-482)
辽宁中医药大学重大重点科研项目团队培育项目(2024-JYTCB-053)。
