【摘要】：目的： 1、明確參麥注射液改善心肌缺血再灌注損傷是否通過調節(jié)鈣調蛋白Phospholamban(PLB)的表達和磷酸化水平來改善細胞內鈣濃度的機理；2、探討人參皂苷對缺血心臟的心功能的改善作用及可能的鈣相關的機理；3、比較參麥注射液復方制劑是否優(yōu)于人參皂苷單純制劑。 方法： 利用新生大鼠原代心肌細胞建立缺氧/復氧(I/R)模型,實驗分為正常培養(yǎng)組(N)、正常培養(yǎng)+參麥組(N+SM)、正常培養(yǎng)+人參總皂苷組(N+TSPG)、正常培養(yǎng)+人參皂苷Rgl組(N+Rgl)、缺氧/復氧組(I/R)、缺氧/復氧組+參麥組(I/R+SM)、缺氧/復氧組+人參總皂苷組(I/R+TSPG)、缺氧/復氧+人參皂苷Rgl組(I/R+Rg1),采用流式細胞儀測定凋亡細胞百分率、細胞內鈣的平均熒光值,實時熒光定量聚合酶式反應(RT-PCR)法測定心肌細胞肌漿網(wǎng)鈣泵(SERCA)和PLB的mRNA表達量,Western Blot法測定心肌細胞SERCA和PLB、磷酸化PLB (p-PLB)的蛋白表達量。 結果： 心肌細胞缺氧／復氧后,PLBmRNA及蛋白表達無顯著變化,p-PLB蛋白表達和SERCA的mRNA及蛋白表達均有不同程度的下降,而I/R+Rgl組較I/R組上述變化無明顯差異(P0.05),I/R+SM組p-PLB蛋白表達、p-PLB/PLB、SERCA mRNA及蛋白表達均明顯升高(P0.01),PLB/SERCA明顯下降(P0.01),I/R+TSPG組SERCA mRNA及蛋白表達均明顯升高(P0.01),PLB/SERCA明顯下降(P0.05),p-PLB蛋白表達、p-PLB/PLB無明顯差異(P0.05)。I/R+SM組較I/R+TSPG組SERCA mRNA及蛋白表達明顯升高(P0.05)、PLB/SERCA明顯降低(P0.05)。I/R+SM組較I／R組凋亡細胞百分率均有不同程度下降(P0.05),I/R+SM組較I/R組細胞內鈣的平均熒光值均有不同程度的降低(P0.01),I/R+TSPG組、I/R+Rgl組細胞凋亡率變化無明顯差異(P0.05),I/R+TSPG組細胞內鈣離子濃度較I/R升高(P0.05),I/R+Rgl無明顯變化。 結論： 1、參麥注射液能通過調節(jié)鈣調蛋白PLB的磷酸化水平來改善細胞內鈣濃度的機理來改善心肌缺血再灌注損傷；2、人參總皂苷可抑制缺血再灌注后心肌細胞內鈣離子濃度,進而改善心功能,此過程可能跟促進SERCA表達有關,但不通過鈣調蛋白PLB磷酸化途徑；3、參麥注射液復方制劑可能優(yōu)于人參皂苷單純制劑。
[Abstract]:Objective: 1. To determine whether Shenmai injection can improve the intracellular calcium concentration by regulating the expression and phosphorylation of calmodulin Phospholamban (PLB) in myocardial ischemia-reperfusion injury. (2) to investigate the effects of ginsenoside on cardiac function of ischemic heart and the possible calcium-related mechanism; (3) to compare whether the compound preparation of Shenmai injection is superior to the simple preparation of ginsenoside. Methods: neonatal rat cardiomyocytes were used to establish hypoxia / reoxygenation (I / R) model. The model was divided into normal culture group (N), normal culture group (N SM), normal cultured ginseng total saponin group (N TSPG),). Normal cultured ginsenoside Rgl group (N Rgl), hypoxia / reoxygenation group, hypoxia / reoxygenation group ginseng wheat group (I / R SM), hypoxia / reoxygenation group Ginseng total saponin group (I / R TSPG),) In hypoxia / reoxygenation ginsenoside Rgl group (I / R Rg1), the percentage of apoptotic cells and the average fluorescent value of intracellular calcium were measured by flow cytometry. The mRNA expression of sarcoplasmic reticulum calcium pump (SERCA) and PLB in cardiomyocytes was measured by real-time fluorescence quantitative polymerase chain reaction (RT-PCR). The protein expressions of SERCA and PLB, phosphorylated PLB (p-PLB) in cardiomyocytes were measured by, Western Blot method. Results: after hypoxia / reoxygenation, the expression of PLBmRNA and protein did not change significantly, but the expression of p-PLB protein and mRNA and protein of SERCA decreased to some extent. However, there was no significant difference between the two groups (P0.05). The expression of p-PLB protein, the expression of p-PLB, mRNA and protein in SM group were significantly increased (P0.01), and the expression of PLB/SERCA was significantly decreased (P0.01), but the expression of p-PLB protein in group I / R Rgl was significantly higher than that in group I / R (P < 0.01). The expression of SERCA mRNA and protein in TSPG group was significantly higher than that in control group (P0.01), while the expression of PLB/SERCA was significantly decreased (P0.05), and the expression of p-PLB protein was significantly decreased. There was no significant difference in p-PLB/PLB (P0.05). The expression of SERCA mRNA and protein in I / R SM group was significantly higher than that in I / R TSPG group (P 0.05). The percentage of apoptotic cells in PLB/SERCA SM group was significantly lower than that in I R group (P0.05), and the percentage of apoptotic cells was significantly decreased (P0.05). The average intracellular Ca ~ (2 +) fluorescence value in I / R SM group was significantly lower than that in I / R group (P0.01). There was no significant difference in apoptosis rate between I / R TSPG group and I / R Rgl group (P 0.05). The intracellular Ca ~ (2 +) concentration in I / R TSPG group was higher than that in I / R group (P 0.05), but there was no significant change in I / R Rgl. Conclusion: 1 Shenmai injection can improve the myocardial ischemia-reperfusion injury by regulating the phosphorylation level of calmodulin PLB to improve the intracellular calcium concentration. 2. Ginsenoside could inhibit intracellular calcium concentration and improve cardiac function after ischemia-reperfusion, which might be related to the promotion of SERCA expression, but not through calmodulin PLB phosphorylation pathway. 3. The compound preparation of Shenmai injection may be superior to the simple preparation of ginsenoside.
【學位授予單位】：浙江大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R542.2

【參考文獻】

相關期刊論文 前10條

1 李穎,崔新明,潘力,栗振寶,汲崇德,杜葵琴;人參果皂甙對失血性休克犬心肌保護作用的電鏡觀察和Ca~(2+)分析[J];白求恩醫(yī)科大學學報;1998年05期

2 張令珍,張鳳和,張鳳海;參麥注射液佐治急性心肌梗死并心力衰竭[J];哈爾濱醫(yī)藥;2003年03期

3 孫文娟,劉潔,呂文偉,葉金梅,李龍云;人參皂苷Rg_2對犬戊巴比妥鈉心力衰竭的影響[J];中國藥理學通報;2003年06期

4 陳彥靜,郝然,黃啟福;參麥注射液對Ang Ⅱ誘導培養(yǎng)心肌細胞凋亡的影響[J];中國病理生理雜志;2004年09期

5 孫益蘭,胡申江,王利宏,周建英;急性心肌梗死大鼠鈣調蛋白的改變及卡維地洛的干預作用[J];中國病理生理雜志;2005年06期

6 郝然;婁金麗;張允嶺;鄭宏;黃啟福;;參麥注射液對缺氧心肌細胞凋亡的影響[J];中國病理生理雜志;2007年04期

7 徐德生,馮怡,周躍華,張曉晨,林曉,鄧海林;麥冬多糖中抗急性心肌缺血活性部位研究[J];中成藥;2004年10期

8 劉潔,孫文娟,呂文偉,葉金梅,李龍云;人參皂苷Rg_2與毒毛旋花子苷K的強心作用及毒性比較[J];中草藥;2001年09期

9 趙昕,李智,曹云鵬,耿慶妍,安麗,湯浩;人參皂甙Rg1對缺氧豚鼠心肌細胞游離鈣濃度降低作用的研究[J];中國醫(yī)科大學學報;2001年06期

10 陳閩軍,吳永江,程翼宇;高效液相色譜用于參麥注射液人參皂苷的含量測定和批次一致性考察[J];中國藥學雜志;2003年08期

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