CAJ | 학술논문

目的探讨活血化瘀中药通脉降糖胶囊(TM)对雪旺细胞(SC)凋亡的影响及机制.方法采用血清药理学方法制备TM含药血清培养液.切取Wistar大鼠双侧坐骨神经,并经消化、离心、培养获得SC.(1)将SC分成正常对照组、SC+TM共培养组(TM组)、SC+晚期糖基化终末产物(AGEs)共培养组(AGEs组)、SC+TM+AGEs共培养组(TM+AGEs组),培养24 h后进行Hoechst 33258和S100免疫荧光染色并于荧光显微镜下计数SC.(2)将SC分成正常血清对照Ⅰ组和TM血清Ⅰ组,培养24 h后应用逆转录聚合酶链反应(RT-PCR)检测肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)、脑源性神经营养因子(BDNF)、神经生长因子(NGF) mRNA表达.(3)将SC分成正常血清对照Ⅱ组和TM血清Ⅱ组,培养48 h后应用ELISA法检测TNF-α、IL-6蛋白表达. 结果 (1)与正常对照组(60.1±3.7)相比,TM组SC数量(82.8±3.8)明显增加,AGEs组SC数量(17.1±1.7)明显减少;TM+AGEs组SC数量(42.3±1.6)较AGEs组明显增加,差异均有统计学意义(P＜0.05).(2)与正常血清对照Ⅰ组相比,TM血清Ⅰ组TNF-α、IL-6 mRNA表达量明显降低,BDNF、NGF mRNA表达无明显变化.(3)与正常血清对照Ⅱ组[TNF-α:(54.7±0.2);IL-6:(955.8±6.8)]相比,TM血清Ⅱ组TNF-α、IL-6蛋白表达[TNF-α:(37.9±0.9);IL-6:(861.3±10.2)]明显降低,差异均有统计学意义(P＜0.05). 结论 TM可能是通过抑制TNF-α、IL-6表达从而明显抑制AGEs诱发的SC凋亡.
목적 탐토활혈화어중약통맥강당효낭(TM)대설왕세포(SC)조망적영향급궤제.방법 채용혈청약이학방법제비TM함약혈청배양액.절취Wistar대서쌍측좌골신경,병경소화、리심、배양획득SC.(1)장SC분성정상대조조、SC+TM공배양조(TM조)、SC+만기당기화종말산물(AGEs)공배양조(AGEs조)、SC+TM+AGEs공배양조(TM+AGEs조),배양24 h후진행Hoechst 33258화S100면역형광염색병우형광현미경하계수SC.(2)장SC분성정상혈청대조Ⅰ조화TM혈청Ⅰ조,배양24 h후응용역전록취합매련반응(RT-PCR)검측종류배사인자-α(TNF-α)、백개소-6(IL-6)、뇌원성신경영양인자(BDNF)、신경생장인자(NGF) mRNA표체.(3)장SC분성정상혈청대조Ⅱ조화TM혈청Ⅱ조,배양48 h후응용ELISA법검측TNF-α、IL-6단백표체. 결과 (1)여정상대조조(60.1±3.7)상비,TM조SC수량(82.8±3.8)명현증가,AGEs조SC수량(17.1±1.7)명현감소;TM+AGEs조SC수량(42.3±1.6)교AGEs조명현증가,차이균유통계학의의(P＜0.05).(2)여정상혈청대조Ⅰ조상비,TM혈청Ⅰ조TNF-α、IL-6 mRNA표체량명현강저,BDNF、NGF mRNA표체무명현변화.(3)여정상혈청대조Ⅱ조[TNF-α:(54.7±0.2);IL-6:(955.8±6.8)]상비,TM혈청Ⅱ조TNF-α、IL-6단백표체[TNF-α:(37.9±0.9);IL-6:(861.3±10.2)]명현강저,차이균유통계학의의(P＜0.05). 결론 TM가능시통과억제TNF-α、IL-6표체종이명현억제AGEs유발적SC조망.
Objective To explore the effect oftongmaijiangtang capsule (TM) on apoptosis and its mechanism in Schwann cells (SCs).Methods The TM contained serum was prepared by using blood serum pharmacological method.Bilateral ischiadic nerves from Wistar rats were sliced,digested and centrifuged to acquire the SCs.(1) These SCs were divided into normal control group,AGEs treatment group,TM treatment group and TM+AGEs treatment group;24 h after each treatment,the SCs were detected by immunofluorescent staining using S100 and DNA-specific fluorescent regent Hoechst 33258,and then,the SCs number was compared under fluorescent microscope.(2) SCs were divided into normal control group Ⅰ and TM treatment group Ⅰ;24 h after treatment,reverse transcription-polymerase chain reaction (RT-PCR) was used to measure the mRNA expressions of tumor necrosis factor-α (TNF-α),interleukin-6 (IL-6),brain derived neurotrophic factor (BDNF) and nerve growth factor (NGF).(3) SCs were divided into normal control group Ⅱ and TM treatment group Ⅱ;48 h after treatment,ELISA was employed to detect the protein expressions of TNF-c and IL-6 in SCs.Results (1) As compared with that in the control group (60.1±3.7),the number of SCs in TM treatment group (82.8±3.8) was statistically increased,and that in AGEs treatment group was significantly decreased (17.1± 1.7,P＜0.05);the number of SCs in TM+AGEs treatment group (42.3±1.6) increased significantly as compared with that in AGEs group (P＜0.05).(2) The mRNA expressions of TNF-o and IL-6 in the TM treatment group Ⅰ obviously decreased as compared with that in the control group Ⅰ;but those of BDNF and NGF in the TM treatment group Ⅰ were similar with those in the control group.(3) The protein expressions of TNF-α and IL-6 in the TM treatment group Ⅱ (54.7±0.2,955.8±6.8) were statistically decreased as compared with those in the control group Ⅱ (37.9±0.9,861.3±10.2,P＜0.05).Conclusion TM could effectively inhibit SCs apoptosis induced by AGEs and significantly decrease the mRNA and protein expressions of TNF-α and IL-6 to reduce the apoptosis of SCs.