建立了反相高效液相色谱法测定多不饱和脂肪酸油脂中VE(VE-α、VE-β、VE-γ、VE-δ)的方法,具体如下:样品经甲醇多次提取,冷冻离心净化后高效液相色谱测定,外标法定量。最佳提取条件:称样质量为0.1 g,甲醇使用量为5 m L、提取次数为2...建立了反相高效液相色谱法测定多不饱和脂肪酸油脂中VE(VE-α、VE-β、VE-γ、VE-δ)的方法,具体如下:样品经甲醇多次提取,冷冻离心净化后高效液相色谱测定,外标法定量。最佳提取条件:称样质量为0.1 g,甲醇使用量为5 m L、提取次数为2次。检出限(LODs)为(0.01~0.06)mg/kg,定量限(LOQs)为(0.04~0.2)mg/kg。4种目标物质在25、50、100 mg/kg的加标水平下的平均回收率在81.4%~107.0%,相对标准偏差2.6%~4.6%(N=6)。该方法结果准确可靠,可用于多不饱和脂肪酸油脂中VE的分析测定。展开更多
A large number of protease inhibitors have been found from leeches,which are essential in various physiological and biological processes.In the curret study,a novel elastase inhibitor was purified and characterized fr...A large number of protease inhibitors have been found from leeches,which are essential in various physiological and biological processes.In the curret study,a novel elastase inhibitor was purified and characterized from the leech of Hirudinaria manillensis,which was named HMEI-A.Primary structure analysis showed that HMEI-A belonged to a new family of proteins.HMEI-A exerted inhibitory effects on elastase and showed potent abilities to inhibit elastase with an inhibition constant(Ki)of 1.69×10^(−8) mol·L^(−1).Further study showed that HMEI-A inhibited the formation of neutrophil extracellular trap(NET).These results suggested that HMEIA from the leech of H.manillensis is a novel elastase inhibitor which can suppress NET formation.It may play a significant role in blood-sucking of leeches and is a potential candidate as an anti-inflammatory agent.展开更多
It is hypothesized that protease inhibitors play an essential role in survival of venomous animals through protecting peptide/protein toxins from degradation by proteases in their prey or predators. However, the biolo...It is hypothesized that protease inhibitors play an essential role in survival of venomous animals through protecting peptide/protein toxins from degradation by proteases in their prey or predators. However, the biological function of protease inhibitors in scorpion venoms remains unknown. In the present study, a trypsin inhibitor was purified and characterized from the venom of scorpion Mesobuthus eupeus, which enhanced the biological activities of crude venom components in mice when injected in combination with crude venom. This protease inhibitor, named Me KTT-1, belonged to Kunitz-type toxins subfamily. Native Me KTT-1 selectively inhibited trypsin with a K_i value of 130 nmol×L^(–1). Furthermore, Me KTT-1 was shown to be a thermo-stable peptide. In animal behavioral tests, Me KTT-1 prolonged the pain behavior induced by scorpion crude venom, suggesting that protease inhibitors in scorpion venom inhibited proteases and protect the functionally important peptide/protein toxins from degradation, consequently keeping them active longer. In conclusion, this was the first experimental evidence about the natural existence of serine protease inhibitor in the venom of scorpion Mesobuthus eupeus, which preserved the activity of venom components, suggests that scorpions may use protease inhibitors for survival.展开更多
文摘建立了反相高效液相色谱法测定多不饱和脂肪酸油脂中VE(VE-α、VE-β、VE-γ、VE-δ)的方法,具体如下:样品经甲醇多次提取,冷冻离心净化后高效液相色谱测定,外标法定量。最佳提取条件:称样质量为0.1 g,甲醇使用量为5 m L、提取次数为2次。检出限(LODs)为(0.01~0.06)mg/kg,定量限(LOQs)为(0.04~0.2)mg/kg。4种目标物质在25、50、100 mg/kg的加标水平下的平均回收率在81.4%~107.0%,相对标准偏差2.6%~4.6%(N=6)。该方法结果准确可靠,可用于多不饱和脂肪酸油脂中VE的分析测定。
基金supported from the National Science Foundation of China(Nos.31630075,21761142002,and 31801975)the Strategic Leading Project and Biological Resource Platform Project from Chinese Academy of Sciences(Nos.KFZDSW-219-2,ZSTH-034 and SAJC201606)+2 种基金the Natural Science Foundation of Yunnan Province(Nos.2019-YT-053 and 202001AT070116)the Social Development Foundation of Yunnan Province(Nos.2019ZF003 and 2018ZF001)Key Research and Development Plan from the Ministry of Science and Technology of China(No.2018YFA0801403).
文摘A large number of protease inhibitors have been found from leeches,which are essential in various physiological and biological processes.In the curret study,a novel elastase inhibitor was purified and characterized from the leech of Hirudinaria manillensis,which was named HMEI-A.Primary structure analysis showed that HMEI-A belonged to a new family of proteins.HMEI-A exerted inhibitory effects on elastase and showed potent abilities to inhibit elastase with an inhibition constant(Ki)of 1.69×10^(−8) mol·L^(−1).Further study showed that HMEI-A inhibited the formation of neutrophil extracellular trap(NET).These results suggested that HMEIA from the leech of H.manillensis is a novel elastase inhibitor which can suppress NET formation.It may play a significant role in blood-sucking of leeches and is a potential candidate as an anti-inflammatory agent.
基金supported by MOST(2013CB911300,2010CB529800)NSFC(Nos.31025025,31025025,U1132601,31200590)+1 种基金CAS(SAJC201308)Yunnan Province(2011CI139,2012BC009)
文摘It is hypothesized that protease inhibitors play an essential role in survival of venomous animals through protecting peptide/protein toxins from degradation by proteases in their prey or predators. However, the biological function of protease inhibitors in scorpion venoms remains unknown. In the present study, a trypsin inhibitor was purified and characterized from the venom of scorpion Mesobuthus eupeus, which enhanced the biological activities of crude venom components in mice when injected in combination with crude venom. This protease inhibitor, named Me KTT-1, belonged to Kunitz-type toxins subfamily. Native Me KTT-1 selectively inhibited trypsin with a K_i value of 130 nmol×L^(–1). Furthermore, Me KTT-1 was shown to be a thermo-stable peptide. In animal behavioral tests, Me KTT-1 prolonged the pain behavior induced by scorpion crude venom, suggesting that protease inhibitors in scorpion venom inhibited proteases and protect the functionally important peptide/protein toxins from degradation, consequently keeping them active longer. In conclusion, this was the first experimental evidence about the natural existence of serine protease inhibitor in the venom of scorpion Mesobuthus eupeus, which preserved the activity of venom components, suggests that scorpions may use protease inhibitors for survival.
