中华糖尿病杂志
中華糖尿病雜誌
중화당뇨병잡지
CHINES JOURNAL OF DLABETES MELLITUS
2012年
10期
630-634
,共5页
胰岛β细胞%钙通道%胰岛素
胰島β細胞%鈣通道%胰島素
이도β세포%개통도%이도소
Islet β cell%Calcium channel%Insulin
目的 通过胰腺原位灌注实验结合钙通道阻断剂的应用探讨L型和P/Q型钙通道在调控大鼠胰岛β细胞胰岛素分泌中的作用.方法 健康雄性SD大鼠24只,采用随机数字表法分为对照组(n＝8)、L型通道阻断组(n=8)和P/Q型通道阻断组(=8)用戊巴比妥钠对大鼠进行腹腔注射麻醉,沿腹正中线做一切口,将胰腺与脾、胃、结肠等组织器官分离,分别在腹主动脉和门静脉上适当的位置插入硅胶导管.各组大鼠均先用37℃改良三羧酸循环4-羟乙基哌嗪乙磺酸缓冲液(modified Krebs-Ringer HEPES)通过腹主动脉插管以1ml/min流速预灌注40 min,随后开始正式灌注实验并收集门静脉插管的流岀液.对照组先用含3.3 mmol/L葡萄糖的Modified Krebs-Ringer HEPES低糖缓冲液对完整的大鼠胰腺灌注10 min,再用含16.7 mmol/L葡萄糖的Modified Krebs-RingerHEPES高糖缓冲液灌注25min；L型通道阻断组先用低糖灌注液灌注10 min,再用50 nmol/L依拉地平-高糖灌注液灌注25 min; P/Q型通道阻断组用低糖灌注液灌注10 min,再用10 nmol/L漏斗网蛛毒素-高糖灌注液灌注25 min各组均从门静脉收集每分钟的流岀液,用放射免疫分析法检测其胰岛素水平.以第1～10分钟为低糖灌注液灌注期,第11～15分钟为第1时相,第16 ～35分钟为第2时相,计算胰岛素分泌率.多组数据比较采用单因素方差分析,进步两两比较采用SNK-q检验.结果 对照组、L型通道阻断组和P/Q型通道阻断组大鼠的体重、空腹血糖值比较差异均无统计学意义(F值分别为1.224、0.377,均P＞0.05).大鼠基础胰岛素分泌率各组间比较差异均无统计学意义(F=0.095,P＞0.05).高糖刺激下,L型通道阻断组的胰岛素1相和2相分泌率及分泌峰值均显著低于对照组和P/Q型通道阻断组[分别为(402 ±24)、(744±32)、(728 ±42)μU/min,F=163.879,P ＜0.01;(323 ±29)、(568±40)、(563 ± 19) μU/min,F=95.043,P＜0.01;(521±43)、(1134±146)、(1083±199) μU/min,F =27.713,P＜0.01];P/Q型通道阻断组的胰岛素1相和2相分泌率及分泌峰值与对照纽比较差异均无统计学意义(t值分别为163.879、95.043、27.713,均P＞0.05).结论 阻断L型钙通道能显著抑制高糖刺激的大鼠胰岛β细胞胰岛素双相分泌,阻断P/Q型钙通道对大鼠胰岛β细胞胰岛素的分泌未见显著影响,表明正常大鼠胰岛β细胞L型钙通道在调控胰岛素双相分泌中起主要作用,而P/Q型钙通道对胰岛素双相分泌的调控并无显著作用.
目的 通過胰腺原位灌註實驗結閤鈣通道阻斷劑的應用探討L型和P/Q型鈣通道在調控大鼠胰島β細胞胰島素分泌中的作用.方法 健康雄性SD大鼠24隻,採用隨機數字錶法分為對照組(n＝8)、L型通道阻斷組(n=8)和P/Q型通道阻斷組(=8)用戊巴比妥鈉對大鼠進行腹腔註射痳醉,沿腹正中線做一切口,將胰腺與脾、胃、結腸等組織器官分離,分彆在腹主動脈和門靜脈上適噹的位置插入硅膠導管.各組大鼠均先用37℃改良三羧痠循環4-羥乙基哌嗪乙磺痠緩遲液(modified Krebs-Ringer HEPES)通過腹主動脈插管以1ml/min流速預灌註40 min,隨後開始正式灌註實驗併收集門靜脈插管的流岀液.對照組先用含3.3 mmol/L葡萄糖的Modified Krebs-Ringer HEPES低糖緩遲液對完整的大鼠胰腺灌註10 min,再用含16.7 mmol/L葡萄糖的Modified Krebs-RingerHEPES高糖緩遲液灌註25min；L型通道阻斷組先用低糖灌註液灌註10 min,再用50 nmol/L依拉地平-高糖灌註液灌註25 min; P/Q型通道阻斷組用低糖灌註液灌註10 min,再用10 nmol/L漏鬥網蛛毒素-高糖灌註液灌註25 min各組均從門靜脈收集每分鐘的流岀液,用放射免疫分析法檢測其胰島素水平.以第1～10分鐘為低糖灌註液灌註期,第11～15分鐘為第1時相,第16 ～35分鐘為第2時相,計算胰島素分泌率.多組數據比較採用單因素方差分析,進步兩兩比較採用SNK-q檢驗.結果 對照組、L型通道阻斷組和P/Q型通道阻斷組大鼠的體重、空腹血糖值比較差異均無統計學意義(F值分彆為1.224、0.377,均P＞0.05).大鼠基礎胰島素分泌率各組間比較差異均無統計學意義(F=0.095,P＞0.05).高糖刺激下,L型通道阻斷組的胰島素1相和2相分泌率及分泌峰值均顯著低于對照組和P/Q型通道阻斷組[分彆為(402 ±24)、(744±32)、(728 ±42)μU/min,F=163.879,P ＜0.01;(323 ±29)、(568±40)、(563 ± 19) μU/min,F=95.043,P＜0.01;(521±43)、(1134±146)、(1083±199) μU/min,F =27.713,P＜0.01];P/Q型通道阻斷組的胰島素1相和2相分泌率及分泌峰值與對照紐比較差異均無統計學意義(t值分彆為163.879、95.043、27.713,均P＞0.05).結論 阻斷L型鈣通道能顯著抑製高糖刺激的大鼠胰島β細胞胰島素雙相分泌,阻斷P/Q型鈣通道對大鼠胰島β細胞胰島素的分泌未見顯著影響,錶明正常大鼠胰島β細胞L型鈣通道在調控胰島素雙相分泌中起主要作用,而P/Q型鈣通道對胰島素雙相分泌的調控併無顯著作用.
목적 통과이선원위관주실험결합개통도조단제적응용탐토L형화P/Q형개통도재조공대서이도β세포이도소분비중적작용.방법 건강웅성SD대서24지,채용수궤수자표법분위대조조(n＝8)、L형통도조단조(n=8)화P/Q형통도조단조(=8)용무파비타납대대서진행복강주사마취,연복정중선주일절구,장이선여비、위、결장등조직기관분리,분별재복주동맥화문정맥상괄당적위치삽입규효도관.각조대서균선용37℃개량삼최산순배4-간을기고진을광산완충액(modified Krebs-Ringer HEPES)통과복주동맥삽관이1ml/min류속예관주40 min,수후개시정식관주실험병수집문정맥삽관적류출액.대조조선용함3.3 mmol/L포도당적Modified Krebs-Ringer HEPES저당완충액대완정적대서이선관주10 min,재용함16.7 mmol/L포도당적Modified Krebs-RingerHEPES고당완충액관주25min；L형통도조단조선용저당관주액관주10 min,재용50 nmol/L의랍지평-고당관주액관주25 min; P/Q형통도조단조용저당관주액관주10 min,재용10 nmol/L루두망주독소-고당관주액관주25 min각조균종문정맥수집매분종적류출액,용방사면역분석법검측기이도소수평.이제1～10분종위저당관주액관주기,제11～15분종위제1시상,제16 ～35분종위제2시상,계산이도소분비솔.다조수거비교채용단인소방차분석,진보량량비교채용SNK-q검험.결과 대조조、L형통도조단조화P/Q형통도조단조대서적체중、공복혈당치비교차이균무통계학의의(F치분별위1.224、0.377,균P＞0.05).대서기출이도소분비솔각조간비교차이균무통계학의의(F=0.095,P＞0.05).고당자격하,L형통도조단조적이도소1상화2상분비솔급분비봉치균현저저우대조조화P/Q형통도조단조[분별위(402 ±24)、(744±32)、(728 ±42)μU/min,F=163.879,P ＜0.01;(323 ±29)、(568±40)、(563 ± 19) μU/min,F=95.043,P＜0.01;(521±43)、(1134±146)、(1083±199) μU/min,F =27.713,P＜0.01];P/Q형통도조단조적이도소1상화2상분비솔급분비봉치여대조뉴비교차이균무통계학의의(t치분별위163.879、95.043、27.713,균P＞0.05).결론 조단L형개통도능현저억제고당자격적대서이도β세포이도소쌍상분비,조단P/Q형개통도대대서이도β세포이도소적분비미견현저영향,표명정상대서이도β세포L형개통도재조공이도소쌍상분비중기주요작용,이P/Q형개통도대이도소쌍상분비적조공병무현저작용.
Objective To study the role of L-type and P/Q-type calcium channels in insulin secretion of rat islet β cell.Methods Healthy SD rats were randomly divided into control group (n =8),L-type channel blocking group (n =8) and P/Q-type channel blocking group (n =8).Sodium pentobarbital was injected into abdominal cavity of the rat.Then a slice was made in its abdomen and separated the pancreas from spleen,stomach,colon and other organs.Finally the silicone catheters were inserted into the abdominal aorta and portal vein on the proper position.The pancreas of rats of the three groups were first perfused with 3.3 nmol/L glucose solution for 10 minutes and then with 16.7 mmol/L glucose solution for 25 minutes.The glucose solution for L-type channel blocking group and P/Q-type channel blocking group contains 50 nmol/L isradipine and 10 nmol/Lω-Agatoxin IVA respectively.Collect the fluid from the portal vein per minute to detect the insulin level by radioimmunoassay.Using SPSS-statistical software package,the experimental data were analyzed through the method of Student Newman Keuls of One-Way-ANOVA.Results The fasting blood glucose and body weight were no significant difference among three groups (F=0.377,1.224,P ＞ 0.05).The basal insulin secretion rates were no significant difference among three groups (F =0.095,P ＞ 0.05).Clucose-evoked insulin secretion rate of the first phase and the second phase,secretion peak value of insulin in L-type channel blocking group were significantly lower than those in the control group respectively [(402 ± 24) vs (744 ± 32) vs (728 ± 42) μU/min,F =163.879,P ＜ 0.01 ; (323±29) vs(568 ±40) vs (563 ± 19) μL/min,F=95.043,P ＜0.01 ;(521 ±43)vs(1134 ± 146)vs (1083 ±199) μU/min,F =27.713,P ＜ 0.01].Clucose-evoked insulin secretion rate of the first phase and the second phase,secretion peak value of insulin were no significantly differences between the P/Q-type channel blocking group and the control group (P＞ 0.05).Conclusions Blocking L-type calcium channel significantly inhibited the biphasic insulin secretion of rat islets β cells.Blocking P/Q-type calcium channel doesn't affect the function of the insulin secretion of the rat islet β cells.It showed that L-type calcium channels of rat islets cells play an important role in biphasic secretion of insulin,but P/Q-type calcium channels are not found to play a role in insulin secretion at any stage.
