Objectives:To investigate the effect on essential hypertension of the topical application of TAT-Cu,Zn-superoxide dismutase(TAT-SOD)at left acupoint Zusanli(ST 36),and to observe whether the change of electrical poten...Objectives:To investigate the effect on essential hypertension of the topical application of TAT-Cu,Zn-superoxide dismutase(TAT-SOD)at left acupoint Zusanli(ST 36),and to observe whether the change of electrical potential difference(EPD)can be related to the change of blood pressure.Methods:Sixteen patients with essential hypertension and 16 healthy subjects were included in the study.EPD between the left acupoints of Yanglingquan(GB 34)and Qiuxu(GB 40)was firstly screened out for the EPD detection.An intracellular superoxide quenching enzyme,TAT-SOD,was topically applied to the acupoint ST 36 within an area of 1 cm^2 once a day,and the influence on EPD was investigated.The dosage applied to TAT-SOD group(n=8)was 0.2 mL of 3000 U/mL TAT-SOD cream prepared by adding purified TAT-SOD to a vehicle cream,while placebo group(n=8)used the vehicle cream instead.The left acupoints of Yanglingquan(GB 34)and Qiuxu(GB 40)were selected for EPD measurement after comparing EPD readings between 5 acupoints on each of all 12 meridians.Results:EPDs between the left acupoints of GB 34 and GB 40 for 16 patients of essential hypertension and 16 healthy subjects were 44.9±6.4 and 5.6±0.9 mV,respectively.Daily application of TAT-SOD for 15 days at ST 36 of essential hypertension patients significantly decreased systolic blood pressure(SBP)and diastolic blood pressure(DBP)of 179.6 and 81.5 mm Hg to 153.1 and 74.1 mm Hg,respectively.Responding to the change in blood pressure,EPD between the left acupoints of GB 34 and GB 40 also declined from 44.4 to 22.8 mV with the same trend.No change was observed with SBP,DBP and EPD between the left acupoints of GB 34 and GB 40 with the daily application of the placebo cream.Conclusion:Enzymatic scavenging of the intracellular superoxide at ST 36 proved to be effective in decreasing SBP and DBP.The results reconfirm the involvement of superoxide anions and its transportation along the meridians,and demonstrate that EPD between acupoints may be an indicator to reflect its functioning status.Moreover,prellmlnary results suggest a close correlatlon between EPD and blood pressure readings,implying a possibility of using EPD as a sensitive parameter for blood pressure and to monitor the effect of antihypertensive treatment.展开更多
目的以重组毕赤酵母发酵液冻干粉为原料,分离纯化超氧化物歧化酶(superoxide dismutase,SOD)与人类Ⅰ型免疫缺陷病毒表达的反式转录激活因子(trans-activator of transcription,TAT)蛋白转导结构域的融合蛋白TATSOD。方法利用乙醇沉淀...目的以重组毕赤酵母发酵液冻干粉为原料,分离纯化超氧化物歧化酶(superoxide dismutase,SOD)与人类Ⅰ型免疫缺陷病毒表达的反式转录激活因子(trans-activator of transcription,TAT)蛋白转导结构域的融合蛋白TATSOD。方法利用乙醇沉淀、大孔树脂柱层析、SP-650离子交换柱层析、TritonX-114相分离法去除TAT-SOD中的色素和细菌内毒素。结果70%乙醇沉降TAT-SOD的酶活力回收率达(98.50±1.71)%,纯化倍数为(1.04±0.02)倍。大孔树脂分离纯化TAT-SOD的效果总体优于SP-650;纯化倍数为(3.07±0.06)倍,略高于SP-650[(2.78±0.05)倍];色素去除率为(95.16±1.65)%,远高于SP-650[(44.73±0.77)%];酶活力回收率为(61.17±1.06)%,略低于SP-650[(69.93±1.21)%]。TritonX-114相分离法纯化TAT-SOD,细菌内毒素去除率为(99.89±1.73)%,酶活力回收率为(95.02±1.66)%。最后获得的TAT-SOD的比活力为(10110±175)U/mg。结论乙醇沉降-大孔树脂柱层析-TritonX-114相分离法对TAT-SOD有良好的分离纯化效果,有望为TAT-SOD工业化分离纯化提供新的选择。展开更多
基金National Key R&D Program of China(No.2016YFD0400202)National Natural Science Foundation of China(No.31500685,30973720)+1 种基金Zhejiang Provincial Natural Science Foundation of China(No.LY16C050002)Start-Up Fund of Research in Quanzhou Normal University(No.G16062)。
文摘Objectives:To investigate the effect on essential hypertension of the topical application of TAT-Cu,Zn-superoxide dismutase(TAT-SOD)at left acupoint Zusanli(ST 36),and to observe whether the change of electrical potential difference(EPD)can be related to the change of blood pressure.Methods:Sixteen patients with essential hypertension and 16 healthy subjects were included in the study.EPD between the left acupoints of Yanglingquan(GB 34)and Qiuxu(GB 40)was firstly screened out for the EPD detection.An intracellular superoxide quenching enzyme,TAT-SOD,was topically applied to the acupoint ST 36 within an area of 1 cm^2 once a day,and the influence on EPD was investigated.The dosage applied to TAT-SOD group(n=8)was 0.2 mL of 3000 U/mL TAT-SOD cream prepared by adding purified TAT-SOD to a vehicle cream,while placebo group(n=8)used the vehicle cream instead.The left acupoints of Yanglingquan(GB 34)and Qiuxu(GB 40)were selected for EPD measurement after comparing EPD readings between 5 acupoints on each of all 12 meridians.Results:EPDs between the left acupoints of GB 34 and GB 40 for 16 patients of essential hypertension and 16 healthy subjects were 44.9±6.4 and 5.6±0.9 mV,respectively.Daily application of TAT-SOD for 15 days at ST 36 of essential hypertension patients significantly decreased systolic blood pressure(SBP)and diastolic blood pressure(DBP)of 179.6 and 81.5 mm Hg to 153.1 and 74.1 mm Hg,respectively.Responding to the change in blood pressure,EPD between the left acupoints of GB 34 and GB 40 also declined from 44.4 to 22.8 mV with the same trend.No change was observed with SBP,DBP and EPD between the left acupoints of GB 34 and GB 40 with the daily application of the placebo cream.Conclusion:Enzymatic scavenging of the intracellular superoxide at ST 36 proved to be effective in decreasing SBP and DBP.The results reconfirm the involvement of superoxide anions and its transportation along the meridians,and demonstrate that EPD between acupoints may be an indicator to reflect its functioning status.Moreover,prellmlnary results suggest a close correlatlon between EPD and blood pressure readings,implying a possibility of using EPD as a sensitive parameter for blood pressure and to monitor the effect of antihypertensive treatment.
文摘目的以重组毕赤酵母发酵液冻干粉为原料,分离纯化超氧化物歧化酶(superoxide dismutase,SOD)与人类Ⅰ型免疫缺陷病毒表达的反式转录激活因子(trans-activator of transcription,TAT)蛋白转导结构域的融合蛋白TATSOD。方法利用乙醇沉淀、大孔树脂柱层析、SP-650离子交换柱层析、TritonX-114相分离法去除TAT-SOD中的色素和细菌内毒素。结果70%乙醇沉降TAT-SOD的酶活力回收率达(98.50±1.71)%,纯化倍数为(1.04±0.02)倍。大孔树脂分离纯化TAT-SOD的效果总体优于SP-650;纯化倍数为(3.07±0.06)倍,略高于SP-650[(2.78±0.05)倍];色素去除率为(95.16±1.65)%,远高于SP-650[(44.73±0.77)%];酶活力回收率为(61.17±1.06)%,略低于SP-650[(69.93±1.21)%]。TritonX-114相分离法纯化TAT-SOD,细菌内毒素去除率为(99.89±1.73)%,酶活力回收率为(95.02±1.66)%。最后获得的TAT-SOD的比活力为(10110±175)U/mg。结论乙醇沉降-大孔树脂柱层析-TritonX-114相分离法对TAT-SOD有良好的分离纯化效果,有望为TAT-SOD工业化分离纯化提供新的选择。
