Why Do Glycosides Of Cistanche Have A Protective Effect On Mitochondria Against Adriamycin?
Mar 12, 2022
Contact: joanna.jia@wecistanche.com / WhatsApp: 008618081934791
Hai Liman, Wuliya, Wang Xiaowen
(Department of pathology, X Xinjiang Medical University, Urumqi 830054, China)
Abstract:
Objective: To determine the stereological parameters change of the cardiac muscle mitochondria in mice on glycosides of Citanche, which protected the cardiac muscle mitochondria against adriamycin ( ADR).
Methods: Every group of cardiac muscle Mitochondria was observed by Transmissionelec- trone microscope author, the volume density ( Vvm), surface density ( SVM ), Qm, numerical density ( Num) of mitochondria of the cells were measured, calculated, and compared with each other.
Results: The cardiac muscle and the mitochondria were swollen, with hyperplasia, smooth endoplasmic reticulum expanded. Secondary lysosome increased, the volume of the nuclei expanded Vvm, Svm, Qm, Num of the cardiac muscle were increased (P < 0. 01). Vvm, Svm, Qm, Num of the GCS cardiac muscle were de-creased to compare with the ADR group (P < 0. 01) the result indicate that were the protective effects of GCs on mitochondria of cardiac muscle against ADR. Conclusion: mitochondria of the cardiac muscle were damaged by the ADR, GCs have some protective role.
Keywords: glycosides of Cistanche; adriamycin; mitochondria of the cardiac muscle; stereology
Adriamycin (ADR) is a broad-spectrum anti-tumor drug, but it has a certain toxic effect on the heart muscle. Cistanche total glycosides are the active ingredients extracted from Cistanche, which have the functions of scavenging free radicals and anti-lipid peroxidation [1, 2]. To explore the toxic effect of doxorubicin on the myocardium and find effective drugs to prevent and treat ADR-induced myocardial toxicity, we conducted electron microscopic observations on the myocardium of mice in the ADR group and the GCs group and carried out a quantitative study on the mitochondrial changes of myocardial cells. From the perspective of ultrastructure, explore the toxic effect of ADR on cardiomyocytes and the protective effect of Cistanche glycosides on its injury.
1 Material and methods
1. 1 Laboratory animals and reagents
N IH mice were provided by the Animal Experiment Center of Xinjiang Institute of Endemic Diseases, and the medical laboratory animal certificate number: No. 16-068. The total glycosides of Cistanche (GCs) are extracted from the salt-producing Cistanche in northern Xinjiang. The main ingredient is phenethyl alcohol glycosides, which are provided by the Department of Phytochemistry, School of Pharmacy, Xinjiang Medical University. Adriamycin (ADR) is Meiji Pharmaceutical Co., Ltd.
1.2 Method
A total of 115 NIH mice were selected, weighing (24±2) g, half male and half female. Randomly divided into 5 groups (23 animals in each group): (1) control group: normal saline (NS) 20 ml/kg; (2) ADR injury group: NS 20 ml/kg; (3) GCsⅠ group: GCs 62. 5 mg/kg; (4) GCsⅡ group: GCs 125.0 mg/kg; (5) GCsⅢ group: GCs 25.0 mg/kg, all the above groups were administered intragastrically, once a day. The ADR injury group, GCsⅠ group, GCsⅡ group, and GCsⅢ group were intraperitoneally injected with ADR (17.5 mg/kg) on the 4th day after the administration of NS and GCs. After 48 hours, the animals were sacrificed and the apical myocardial tissue was quickly taken, fixed with 4% glutaraldehyde and 1% acid, acetone shaved and dehydrated, embedded in Epon 812, double stained with lead and uranium, and observed under JEM-100CXⅡ transmission electron microscope.
1. 3 Quantitative analysis of stereology [3]
A quantitative study of cardiomyocyte mitochondria of animals in each experimental group was carried out, that is, in the ultra-thin sections of the embedded blocks in each group, 10 different fields of view were taken at 13,000 times, and a total of 100 fields of view were taken in each group of animals, and then according to the body. The stereology grid measurement method and calculation formula respectively calculate the volume density (Vvm), surface area density (SVM), mitochondrial specific surface area (Qm), and mitochondrial area number density (Num) of cardiomyocyte mitochondria.
1. 4 Statistical processing
Use PEM S software to perform statistical analysis on this group of data, using analysis of variance, test level α = 0.05
2 results
2. 1 Electron microscopic observation results of cardiomyocytes
Control group: Myocardial cell membranes are intact, myofibrils are arranged in order, sarcomere structure is clear, mitochondria are arranged regularly under myofilaments, the structure is normal, and the nuclear structure is normal (Figure 1). ADR injury group: There are obvious changes in the ultrastructure of cardiomyocytes, swelling of cardiomyocytes, diffuse swelling and hyperplasia of mitochondria, sparse ridge arrangement, rupture, vacuolar degeneration, reduced matrix density, sarcoplasmic reticulum expansion, and secondary lysosomes Increase, the nuclear volume increases, and the perinuclear space widens (Figure 2). GCsⅠ group: Myocardial cell swelling and mitochondrial swelling were alleviated, the matrix density was increased, individual mitochondrial cristae still changed, and secondary lysosomes in the cytoplasm increased (Figure 3). GCsⅢ group: Myocardial cell structure returned to normal, myofibrils arranged neatly, sarcomere structure was normal, and mitochondrial structure returned to normal (Figure 4).
2.2 Stereology results of mitochondria in cardiomyocytes
The results of the analysis of variance showed that the Vvm, Svm, Qm, Num of the ADR injury group were significantly different from the control group (P <0.01); the Vvm, Svm, Qm, Num of the GCsⅡ and Ⅲ group were compared with the ADR injury group The difference is statistically significant (P <0.01), see Table 1.
Table 1 Stereological analysis of mitochondria in cardiomyocytes of each experimental group
Note: Comparison between ADR group and control group, * P <0.01; Comparison between GCs group and ADR group, # P <0.01
3 Discussion
Many research data show that doxorubicin has a toxic effect on the myocardium. This study found that after 48 hours of intraperitoneal injection of doxorubicin (17.5 mg/kg) in mice, the mouse cardiomyocytes have obvious ultrastructural changes, which are manifested by swelling of cardiomyocytes, diffuse swelling, and hyperplasia of mitochondria. Vacuole degeneration, sarcoplasmic reticulum expansion; secondary lysosomes increased. The swelling of myocardial cells in the GCsⅠ group was reduced, the swelling of mitochondria was also reduced, and the matrix density increased. The ultrastructure of cardiomyocytes in GCsⅡ and Ⅲ groups returned to normal, and the mitochondrial structure also returned to normal. Regarding the mechanism of ADR-induced myocardial toxicity, the current tendency is toward the "lipid peroxidation theory". The research results of Wu Liya et al. [4] showed that doxorubicin significantly reduced the activities of SOD and Se-GSH-Px, the content of lipid peroxidation product M DA, and the activity of serum CPK. Changes in the activity of these enzymes reduce the damage of cardiomyocytes against lipid peroxides and damage the cell biofilm. Each dose group of GCs can increase the activities of SOD and Se-GSH-Px in the myocardial free radical scavenging enzymes of ADR-injured mice, reduce the content of M DA, and reduce the release of CPK, thereby increasing the activity of myocardial free radical scavenging enzymes, and reducing the ADR-induced production. Lipid peroxide damage.
The results of this study showed that Vvm, Svm, QM, and Num in the ADR group were significantly higher than those in the control group (P <0.01), indicating that ADR can cause swelling and proliferation of myocardial mitochondria and impair mitochondrial function. Compared with the ADR group, Vvm, Svm, Qm, and Num in GCsⅡ and Ⅲ groups were significantly reduced (P <0.01), indicating that GCs have a certain effect on the mitochondrial damage of cardiomyocytes induced by ADR. This is because ADR is converted into semiquinone after entering the cardiomyocytes. ADR acts on oxygen molecules and generates excessive oxygen free radicals, which causes cardiomyocyte damage, and GCs can effectively scavenge oxygen free radicals [5], thereby resisting the effects of free radical damage.
glycosides of Cistanche
References
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[2] Li Linlin, Wang Xiaowen, Wang Xuefei, et al. Anti-lipid peroxidation and anti-radiation effects of total glycosides of Cistanche [J]. Chinese Journal of Chinese Herbal Medicine, 1997, 22(6): 364-367.
[3] Hang Zhenrong, Cai Wenqin. The application of electron microscopy in clinical medicine [M]. Chongqing Press, 1988. 74-101.
[4] Wu Liya Yiming, Wang Xiaowen, Wang Xuefei, et al. Protective effect of Cistanche glycosides on doxorubicin-induced myocardial toxicity in mice [J]. Journal of Xinjiang Medical University, 2003, 26 (1): 28-30.
[5] Wu Yuling, Xu Guangyuan. Experimental study of doxorubicin-induced acute cardiotoxicity in mice and the protective effect of Vito [J]. Journal of Dalian Medical University, 1991, 13(1): 22-27.
From： Journal of Xinjiang Medical University JOURNAL OF XIN JIANG MEDICAL UNIVERSITY 2004 Apr., 27( 2)
About the author: He Liman Yilahun (1963-), female (Uyghur), bachelor, lecturer, research direction: cell ultramicropathology.
