Oxaliplatin(OXA) is a widely used chemotherapeutic agent whose clinical utility is limited by OXA-induced peripheral neuropathy(OIPN). Sarco/endoplasmic reticulum Ca^(2+)-ATPase(SERCA) transports Ca^(2+) from the cyto...Oxaliplatin(OXA) is a widely used chemotherapeutic agent whose clinical utility is limited by OXA-induced peripheral neuropathy(OIPN). Sarco/endoplasmic reticulum Ca^(2+)-ATPase(SERCA) transports Ca^(2+) from the cytoplasm into the endoplasmic reticulum(ER), thereby maintaining intracellular Ca^(2+) homeostasis. Schefflera kwangsiensis Merr. ex H.L. Li(SKM) is traditionally used to treat neuropathic pain conditions such as trigeminal neuralgia and sciatica, and its active component Schekwanglupaside C has been identified as a potent SERCA activator. In this study, an OIPN mouse model was established by intraperitoneal administration of OXA(4 mg·kg^(-1)) on days 1, 2, 8, 9, 15, and 16. SERCA2b mRNA and protein expression in dorsal root ganglia(DRG) were evaluated by quantitative polymerase chain reaction(qPCR) and immunofluorescence. Mechanical allodynia was assessed using the Von Frey test.DRG neuronal excitability was examined by whole-cell current-clamp recordings, whereas oxidative stress and neuronal apoptosis/necrosis were assessed using the reactive oxygen species(ROS)-sensitive probe 2',7'-dichlorofluorescin diacetate(H2 DCFDA) and fluorescein isothiocyanate(FITC)/propidium iodide(PI) dual staining. This study identifies SERCA2b as a novel therapeutic target for OIPN. We observed that SERCA2b mRNA and protein levels were significantly downregulated during OIPN progression. Treatment with the SERCA agonist CDN1163(CDN), the ethyl acetate extract of SKM(SKM.Ext), or duloxetine(DLX) attenuated neuronal pathology, restored DRG neuron soma diameter, and reduced the expression of proinflammatory cytokines interleukin-1β(IL-1β) and tumor necrosis factor α(TNF-α). Pre-incubation of DRG neurons with CDN1163 or SKM.Ext for 1 h significantly attenuated OXA-induced hyperexcitability and reduced the abnormal increase in voltage-gated sodium channel(VGSC) current density. Inhibition of oxidative stress with N-acetyl-L-cysteine(NAC) significantly restored SERCA expression in OIPN, indicating that oxidative stress downregulates SERCA2b in DRG. Collectively, these findings demonstrate that activation of SERCA2b by CDN1163 or Schefflera kwangsiensis extract enhances SERCA2b expression, reduces DRG neuronal sensitization, and alleviates OIPN. This work supports SERCA2b as a novel therapeutic target for OXA-induced neuropathy and expands the potential clinical analgesic indications of Schefflera kwangsiensis.展开更多
目的:研究Ca2+离子对非CaBPs蛋白类过敏原二级结构和抗原活性的影响。方法:以重组榛子主要过敏原Cor h 1为研究对象,用圆二色谱(CD)研究了系列浓度的Ca2+和EDTA各自存在情况下,不同蛋白浓度溶液中rCor h 1二级结构的变化及规律,并以榛...目的:研究Ca2+离子对非CaBPs蛋白类过敏原二级结构和抗原活性的影响。方法:以重组榛子主要过敏原Cor h 1为研究对象,用圆二色谱(CD)研究了系列浓度的Ca2+和EDTA各自存在情况下,不同蛋白浓度溶液中rCor h 1二级结构的变化及规律,并以榛子过敏患者血清为一抗,用ELISA实验分析了Ca2+和EDTA对rCor h 1抗原活性的影响。结果:在高浓度rCor h 1溶液中Ca2+的加入可以引起椭圆率的增加,而在低浓度的rCor h 1溶液中则引起椭圆率的降低;EDTA在高浓度和低浓度的rCor h 1溶液中均能引起rCor h 1二级结构相对含量的降低;椭圆率增加(或降低)的幅度与Ca2+、EDTA终浓度成正比。ELISA实验表明Ca2+和EDTA都能显著降低rCor h 1的抗原活性。结论:Ca2+离子可以显著影响rCor h 1的二级结构,降低其抗原活性,为非CaBPs蛋白类低致敏原的研究奠定了较好的前期研究基础。展开更多
基金supported by the Natural Science Foundation of Jiangsu Province of China (No. BK20221054)the National Natural Science Foundation of China (Nos. 82204653, 82373929, and82100585)+3 种基金the Major Program of Jiangsu Provincial Administration for Market Regulation (No. KJ2024014)“Double First-Class” University Project (No. CPU2022QZ30)the Open Fund Project of State Key Laboratory on Technologies for Chinese Medicine Pharmaceutical Process Control and Intelligent Manufacture (No.SKL2024Z0104)Xizang Autonomous Region Science and Technology Plan Project Key Project (No. XZ202301ZY0014G)。
文摘Oxaliplatin(OXA) is a widely used chemotherapeutic agent whose clinical utility is limited by OXA-induced peripheral neuropathy(OIPN). Sarco/endoplasmic reticulum Ca^(2+)-ATPase(SERCA) transports Ca^(2+) from the cytoplasm into the endoplasmic reticulum(ER), thereby maintaining intracellular Ca^(2+) homeostasis. Schefflera kwangsiensis Merr. ex H.L. Li(SKM) is traditionally used to treat neuropathic pain conditions such as trigeminal neuralgia and sciatica, and its active component Schekwanglupaside C has been identified as a potent SERCA activator. In this study, an OIPN mouse model was established by intraperitoneal administration of OXA(4 mg·kg^(-1)) on days 1, 2, 8, 9, 15, and 16. SERCA2b mRNA and protein expression in dorsal root ganglia(DRG) were evaluated by quantitative polymerase chain reaction(qPCR) and immunofluorescence. Mechanical allodynia was assessed using the Von Frey test.DRG neuronal excitability was examined by whole-cell current-clamp recordings, whereas oxidative stress and neuronal apoptosis/necrosis were assessed using the reactive oxygen species(ROS)-sensitive probe 2',7'-dichlorofluorescin diacetate(H2 DCFDA) and fluorescein isothiocyanate(FITC)/propidium iodide(PI) dual staining. This study identifies SERCA2b as a novel therapeutic target for OIPN. We observed that SERCA2b mRNA and protein levels were significantly downregulated during OIPN progression. Treatment with the SERCA agonist CDN1163(CDN), the ethyl acetate extract of SKM(SKM.Ext), or duloxetine(DLX) attenuated neuronal pathology, restored DRG neuron soma diameter, and reduced the expression of proinflammatory cytokines interleukin-1β(IL-1β) and tumor necrosis factor α(TNF-α). Pre-incubation of DRG neurons with CDN1163 or SKM.Ext for 1 h significantly attenuated OXA-induced hyperexcitability and reduced the abnormal increase in voltage-gated sodium channel(VGSC) current density. Inhibition of oxidative stress with N-acetyl-L-cysteine(NAC) significantly restored SERCA expression in OIPN, indicating that oxidative stress downregulates SERCA2b in DRG. Collectively, these findings demonstrate that activation of SERCA2b by CDN1163 or Schefflera kwangsiensis extract enhances SERCA2b expression, reduces DRG neuronal sensitization, and alleviates OIPN. This work supports SERCA2b as a novel therapeutic target for OXA-induced neuropathy and expands the potential clinical analgesic indications of Schefflera kwangsiensis.
文摘目的:研究Ca2+离子对非CaBPs蛋白类过敏原二级结构和抗原活性的影响。方法:以重组榛子主要过敏原Cor h 1为研究对象,用圆二色谱(CD)研究了系列浓度的Ca2+和EDTA各自存在情况下,不同蛋白浓度溶液中rCor h 1二级结构的变化及规律,并以榛子过敏患者血清为一抗,用ELISA实验分析了Ca2+和EDTA对rCor h 1抗原活性的影响。结果:在高浓度rCor h 1溶液中Ca2+的加入可以引起椭圆率的增加,而在低浓度的rCor h 1溶液中则引起椭圆率的降低;EDTA在高浓度和低浓度的rCor h 1溶液中均能引起rCor h 1二级结构相对含量的降低;椭圆率增加(或降低)的幅度与Ca2+、EDTA终浓度成正比。ELISA实验表明Ca2+和EDTA都能显著降低rCor h 1的抗原活性。结论:Ca2+离子可以显著影响rCor h 1的二级结构,降低其抗原活性,为非CaBPs蛋白类低致敏原的研究奠定了较好的前期研究基础。
