Background:Neuropathic pain(NP)has a long course and is difficult to treat,causing great physical suffering and psychological burden to patients.Unfortunately,the pathogenesis of NP is still poorly understood.The occu...Background:Neuropathic pain(NP)has a long course and is difficult to treat,causing great physical suffering and psychological burden to patients.Unfortunately,the pathogenesis of NP is still poorly understood.The occurrence and progression of NP are influenced by microRNA(miRNA).It has been reported that Tuina may effectively alleviate NP,however,the mechanisms related to miRNA-mediated Tuina for NP are still unexplored.Methods:To generate NP,a rat model of chronic constriction injury(CCI)was developed.Rats’pain thresholds are studied through pain behaviour tests.The effect of Tuina on the structure and morphology of neurones in the spinal dorsal horn(SDH)of rats with neuropathological pain was examined using HE staining.The levels of miRNA in the SDH of rats with a CCI model were studied using RNA sequencing(RNA-seq)and bioinformatics analysis.The top three miRNAs with the highest fold changes were chosen for qRT-PCR detection to confirm the validity of the RNA-seq results.Results:On the 4th day after CCI surgery,the paw withdrawal threshold(PWT)of the CCI+Tuina group was considerably higher than that of the CCI group(P<0.001).Furthermore,there was a significant increase in the paw withdrawal latency(PWL)of the CCI+Tuina group from day 4 to day 17 after the CCI surgery(P<0.001).Differentially expressed miRNAs in the SDH of three groups of rats were examined using RNA-seq technology.Seven miRNAs were found to intersect in total.The Kyoto Encyclopaedia of Genes and Genomes(KEGG)and Gene Ontology(GO)both anticipated the possible roles of the miRNAs.The RNA-seq results matched the changes in the levels of miR-383-3p(P<0.01),miR-183-3p(P<0.01),and miR-184(P<0.01).Conclusion:CCI-induced NP in rats leads to changes in miRNA expression in the SDH,and Tuina may alleviate NP by modulating the expression pattern of miRNAs and their underlying mechanisms.These findings may provide new targets for future NP treatment.展开更多
Objective:Preventing the transition from acute to chronic pain(pain transition)is a new strategy for treating chronic pain.The present study aimed to investigate the role of K+-Cl−Cotransporter Isoform 2(KCC2)andγ-am...Objective:Preventing the transition from acute to chronic pain(pain transition)is a new strategy for treating chronic pain.The present study aimed to investigate the role of K+-Cl−Cotransporter Isoform 2(KCC2)andγ-aminobutyric acid receptor type A(GABAAR)in the spinal cord dorsal horn(SCDH)in pain transition and the intervention effect of electroacupuncture(EA),and to understand the mechanism of EA in preventing acute and chronic pain transition in the spinal center.Methods:A rat model of hyperalgesic priming(HP)was established by injecting carrageenan(Car)into the plantar area of rats,followed by the injection of prostaglandin E2(PGE2)into the dorsal foot 7 days later.The GABAAR agonist(muscimol)and KCC2 activator(CLP257)were intrathecally injected for three consecutive days after PGE2 injection.EA was applied at a frequency of 2/100 Hz to the bilateral foot Zusanli(ST36)and Kunlun(BL60).A von Frey filament was used to detect the pain threshold in each group of rats.Western blotting(WB)and immunofluorescence(IF)were used to detect GABAAR and KCC2 expression in each rats group.By combining EA intervention with a KCC2 inhibitor(VU0240551),we explored the mechanism of pain transition of EA regulation of GABAAR and KCC2 expression in SCDH.Results:The HP model was established by injecting mice with Car/PGE2.Compared to the normal saline(NS)+NS and NS+PGE2 groups,the pain threshold of the Car+PGE2 group decreased significantly 48 hours after PGE2 injection(P<0.01).The WB results indicated that intrathecal injection of a GABAAR agonist upregulated GABAAR expression in the SCDH of HP model rats(P<0.05).WB and IF results revealed that intrathecal injection of the KCC2 activator significantly increased GABAAR and KCC2 expression in the SCDH of HP model rats(P<0.01)and that GABAAR and KCC2 were co-expressed in the same SCDH cells.Compared to the Car+PGE2 group,EA intervention significantly increased MWTs from 48 to 72 hours after the first injection and 4,24,and 48 hours after the second injection(P<0.01).EA upregulated GABAAR and KCC2 expression in the SCDH of rats with HP(P<0.05).Intrathecal injection of the KCC2 inhibitor blocked the analgesic effect of EA in HP model rats(P<0.01).Conclusions:In SCDH,KCC2 expression was downregulated,causing downregulation of GABAAR expression and resulting in pain transition.EA upregulates KCC2 and GABAAR expression and prevents pain transition.展开更多
Highlights●CRISPR/Cas9 RNP complex-based strategy demonstrates robustness and accuracy in generating gene-edited sheep.●Sheep horn development remains unaffected by partial RXFP2 knockout.●Partial RXFP2 knockout re...Highlights●CRISPR/Cas9 RNP complex-based strategy demonstrates robustness and accuracy in generating gene-edited sheep.●Sheep horn development remains unaffected by partial RXFP2 knockout.●Partial RXFP2 knockout results in unilateral cryptorchidism in sheep.展开更多
Persistent postsurgical pain is a major clinical concern,especially in the aging population,who represent a growing proportion of surgical patients.Although age is a known pain risk factor,the mechanisms driving age-r...Persistent postsurgical pain is a major clinical concern,especially in the aging population,who represent a growing proportion of surgical patients.Although age is a known pain risk factor,the mechanisms driving age-related vulnerability to chronic postoperative pain remain poorly understood.This study aims to investigate how aging influences the resolution of postoperative pain and to elucidate the roles of microglial activation and synaptic remodeling in the spinal dorsal horn.A plantar incision model in young(3-month-old)and aged(18-month-old)male and female mice was used to mimic postoperative pain conditions.Mechanical and thermal hypersensitivity at various postoperative intervals were assessed by von Frey and Hargreaves tests.Microglial activation and inhibitory/excitatory synaptic densities in the spinal dorsal horn were evaluated using immunofluorescence and 3D reconstruction with Imaris software.On postoperative day(POD)3,both age groups exhibited reduced pain thresholds on the ipsilateral side,along with microglial activation in the dorsal horn.On POD 7,pain thresholds in young mice had returned to baseline with no significant microglial activation,while aged mice showed sustained reduction in pain thresholds,continuous microglial activation,and significant loss of inhibitory synapses without detectable changes in excitatory synapse density.These findings are consistent across both sexes,with no sex-related differences.Collectively,these results suggest that aging is associated with persistent postoperative pain,which correlates with microglial activation and inhibitory synapse loss.These insights advance our understanding of age-related pain vulnerability and may inform the development of more effective,targeted,and age-specific therapeutic strategies to prevent or alleviate persistent postoperative pain in elderly patients.展开更多
基金supported by the Project of National Natural Science Foundation of China(No.82174523,No.82205303)Fujian Provincial Health Commission Traditional Chinese Medicine Science and Technology Plan Project(No.2025YBB010)Natural Science Foundation of Fujian Province(No.2023J06037,No.2024J01141).
文摘Background:Neuropathic pain(NP)has a long course and is difficult to treat,causing great physical suffering and psychological burden to patients.Unfortunately,the pathogenesis of NP is still poorly understood.The occurrence and progression of NP are influenced by microRNA(miRNA).It has been reported that Tuina may effectively alleviate NP,however,the mechanisms related to miRNA-mediated Tuina for NP are still unexplored.Methods:To generate NP,a rat model of chronic constriction injury(CCI)was developed.Rats’pain thresholds are studied through pain behaviour tests.The effect of Tuina on the structure and morphology of neurones in the spinal dorsal horn(SDH)of rats with neuropathological pain was examined using HE staining.The levels of miRNA in the SDH of rats with a CCI model were studied using RNA sequencing(RNA-seq)and bioinformatics analysis.The top three miRNAs with the highest fold changes were chosen for qRT-PCR detection to confirm the validity of the RNA-seq results.Results:On the 4th day after CCI surgery,the paw withdrawal threshold(PWT)of the CCI+Tuina group was considerably higher than that of the CCI group(P<0.001).Furthermore,there was a significant increase in the paw withdrawal latency(PWL)of the CCI+Tuina group from day 4 to day 17 after the CCI surgery(P<0.001).Differentially expressed miRNAs in the SDH of three groups of rats were examined using RNA-seq technology.Seven miRNAs were found to intersect in total.The Kyoto Encyclopaedia of Genes and Genomes(KEGG)and Gene Ontology(GO)both anticipated the possible roles of the miRNAs.The RNA-seq results matched the changes in the levels of miR-383-3p(P<0.01),miR-183-3p(P<0.01),and miR-184(P<0.01).Conclusion:CCI-induced NP in rats leads to changes in miRNA expression in the SDH,and Tuina may alleviate NP by modulating the expression pattern of miRNAs and their underlying mechanisms.These findings may provide new targets for future NP treatment.
基金supported by the Natural Science Foundation of Zhejiang Province(No.LY23H270007)。
文摘Objective:Preventing the transition from acute to chronic pain(pain transition)is a new strategy for treating chronic pain.The present study aimed to investigate the role of K+-Cl−Cotransporter Isoform 2(KCC2)andγ-aminobutyric acid receptor type A(GABAAR)in the spinal cord dorsal horn(SCDH)in pain transition and the intervention effect of electroacupuncture(EA),and to understand the mechanism of EA in preventing acute and chronic pain transition in the spinal center.Methods:A rat model of hyperalgesic priming(HP)was established by injecting carrageenan(Car)into the plantar area of rats,followed by the injection of prostaglandin E2(PGE2)into the dorsal foot 7 days later.The GABAAR agonist(muscimol)and KCC2 activator(CLP257)were intrathecally injected for three consecutive days after PGE2 injection.EA was applied at a frequency of 2/100 Hz to the bilateral foot Zusanli(ST36)and Kunlun(BL60).A von Frey filament was used to detect the pain threshold in each group of rats.Western blotting(WB)and immunofluorescence(IF)were used to detect GABAAR and KCC2 expression in each rats group.By combining EA intervention with a KCC2 inhibitor(VU0240551),we explored the mechanism of pain transition of EA regulation of GABAAR and KCC2 expression in SCDH.Results:The HP model was established by injecting mice with Car/PGE2.Compared to the normal saline(NS)+NS and NS+PGE2 groups,the pain threshold of the Car+PGE2 group decreased significantly 48 hours after PGE2 injection(P<0.01).The WB results indicated that intrathecal injection of a GABAAR agonist upregulated GABAAR expression in the SCDH of HP model rats(P<0.05).WB and IF results revealed that intrathecal injection of the KCC2 activator significantly increased GABAAR and KCC2 expression in the SCDH of HP model rats(P<0.01)and that GABAAR and KCC2 were co-expressed in the same SCDH cells.Compared to the Car+PGE2 group,EA intervention significantly increased MWTs from 48 to 72 hours after the first injection and 4,24,and 48 hours after the second injection(P<0.01).EA upregulated GABAAR and KCC2 expression in the SCDH of rats with HP(P<0.05).Intrathecal injection of the KCC2 inhibitor blocked the analgesic effect of EA in HP model rats(P<0.01).Conclusions:In SCDH,KCC2 expression was downregulated,causing downregulation of GABAAR expression and resulting in pain transition.EA upregulates KCC2 and GABAAR expression and prevents pain transition.
基金supported by the National Key Research and Development Program of China(2022YFD1300200)the National Natural Science Foundation of China(32161143010,32202646,and 32272848)+2 种基金the China Agriculture Research System(CARS-39)the Key Special Project of Ningxia Science and Technology Department,China(2021BEF02024)the local grants,China(NXTS2021-001,2022GD-TSLD-46,NK2022010207,and NXTS2022-001)。
文摘Highlights●CRISPR/Cas9 RNP complex-based strategy demonstrates robustness and accuracy in generating gene-edited sheep.●Sheep horn development remains unaffected by partial RXFP2 knockout.●Partial RXFP2 knockout results in unilateral cryptorchidism in sheep.
基金supported by the National Natural Science Foundation of China(No.82401445 and 82271249)the China Postdoctoral Science Foundation(No.2024M752251)+3 种基金the Postdoctoral Fellowship Program of CPSF(No.GZC20241141)the Sichuan Science and Technology Program(No.2024NSFSC1636 and 2025ZNSFSC1645)the Postdoctoral Research Fund of West China Hospital of Sichuan University(No.2024HXBH013)1-3-5 Project for Disciplines of Excellence of West China Hospital of Sichuan University(No.ZYYC23002)。
文摘Persistent postsurgical pain is a major clinical concern,especially in the aging population,who represent a growing proportion of surgical patients.Although age is a known pain risk factor,the mechanisms driving age-related vulnerability to chronic postoperative pain remain poorly understood.This study aims to investigate how aging influences the resolution of postoperative pain and to elucidate the roles of microglial activation and synaptic remodeling in the spinal dorsal horn.A plantar incision model in young(3-month-old)and aged(18-month-old)male and female mice was used to mimic postoperative pain conditions.Mechanical and thermal hypersensitivity at various postoperative intervals were assessed by von Frey and Hargreaves tests.Microglial activation and inhibitory/excitatory synaptic densities in the spinal dorsal horn were evaluated using immunofluorescence and 3D reconstruction with Imaris software.On postoperative day(POD)3,both age groups exhibited reduced pain thresholds on the ipsilateral side,along with microglial activation in the dorsal horn.On POD 7,pain thresholds in young mice had returned to baseline with no significant microglial activation,while aged mice showed sustained reduction in pain thresholds,continuous microglial activation,and significant loss of inhibitory synapses without detectable changes in excitatory synapse density.These findings are consistent across both sexes,with no sex-related differences.Collectively,these results suggest that aging is associated with persistent postoperative pain,which correlates with microglial activation and inhibitory synapse loss.These insights advance our understanding of age-related pain vulnerability and may inform the development of more effective,targeted,and age-specific therapeutic strategies to prevent or alleviate persistent postoperative pain in elderly patients.
