Anti-lipid peroxidation and anti-radiation effects of Total Glycoside of Cistanche
Mar 15, 2022
Contact: joanna.jia@wecistanche.com / WhatsApp: 008618081934791
Li Linlin, Wang Xiaowen, Wang Xuefei, Mu Hu Yati, Du Niansheng
[Abstract]: Total Glycoside of Cistanche (GCs) can significantly enhance the SOD activity of mouse red blood cells, reduce the serum MDA content, increase the DNA and RNA content in liver and kidney tissues, and can significantly promote the damage caused by 60Co irradiation. The restoration of SOD activity in mouse red blood cells and the content of nucleic acid in the spleen reduces liver MDA content, suggesting that the protective effect of GCs (Total Glycoside of Cistanche) on nucleic acids and radioprotection is closely related to its anti-lipid peroxidation effect.
Keywords: Total Glycoside of Cistanche, DNA, RNA, anti-lipid peroxidation, radiation protection
Cistanche has the effects of nourishing the vital energy, nourishing essence, and strengthening yang. In the anti-aging and prolonging ancient prescriptions of the past, the probability of appearance is second only to ginseng. Recent literature reports that Cistanche can enhance the immune function of the body (1); promote memory and improve intelligence; regulate the nervous system, improve sexual function and anti-aging (2). The crude preparation of Cistanche can enhance the SOD activity of mouse red blood cells; significantly reduce the content of myocardial lipofuscin (3); improve the DNA synthesis ability of animals with yang deficiency, and significantly extend the lifespan of fruit flies (4). However, there is no report on the pharmacological effects of GCs (Total Glycoside of Cistanche) at home and abroad. This study aims to explore the protective effect of GCs (Total Glycoside of Cistanche) on nucleic acids, radiation protection, and anti-lipid peroxidation.
Total Glycoside of Cistanche anti-lipid peroxidation
1 Material and methods
1.1 Drugs and reagents
Total Glycoside of Cistanche, extracted from Cistanche Salsa (Cistanche salsa C.A.Mey) from Northern Xinjiang, the main ingredient is Phnylethanoild Glycoside of Cistanche, and the extraction method is based on Japanese literature (Patented Publication (A) Sho 63-198627) Proceeding, provided by Professor Du Niansheng, Department of Plant Chemistry, Department of Pharmacy. Thiobarbituric acid, produced by Shanghai Reagent No. 1 Factory; DNA and RNA standard products were provided by Shanghai Milk Company and Institute of Biochemistry, Chinese Academy of Sciences, and both were tested for purity according to the phosphorus determination method [5].
1.2 Anti-lipid peroxidation effect of GCs (Total Glycoside of Cistanche)
110 NIH mice, weighing (22.5 ± 2.1) g, both male and female, were randomly divided into 4 groups: the control group was given normal saline (NS) 20ml·kg -1·d-1 gavage (ig); GCs (Total Glycoside of Cistanche) group, GCs (Total Glycoside of Cistanche) 62.5, 125, 250 mg·kg -1 day -1, ig were given for 18 days each, after the last administration for 2 hrs, the eyeballs were taken out and 50 μl of blood was taken to test the SOD in the red blood cells Vitality; the remaining blood to prepare serum, measure the serum MDA content; quickly take the heart, liver, brain, kidney and other organs, rinse with ice normal saline, prepare each tissue homogenate, red blood cell SOD extract is prepared according to literature (6). DNA and RNA extract preparation: take the heart, weigh the liver, brain, and kidney organs, cut them into a grinder, add 2ml, 15% trichloroacetic acid, grind it into a homogenate in an ice water bath, and transfer to centrifugation Tube, rinse the grinder bowl with 2ml 15% trichloroacetic acid for a total of 3 times, transfer all the washing liquid into the centrifuge tube, centrifuge 4000r·min-1 for 10min, discard the supernatant. After adding 2ml of absolute ethanol to the precipitation and mixing, place it in a water bath at 90 ℃ and heat it for 2 min. After cooling, it is centrifuged at 4000 r·min-1 for 10 min, and the supernatant is discarded. Add 8ml of 5% trichloroacetic acid to the precipitation and mix well, put it in a water bath at 90 ℃ for 15min, centrifuge after cooling, 4000r·min-1 for 10min, pour the supernatant into a 10ml volumetric flask and add 1ml to the precipitation 5% trichloroacetic acid, mix well, centrifuge, 4000r·min-1, 5min, and then pour the supernatant into a 10ml volumetric flask, dilute the volume with distilled water, and mix well. The DNA and RNA extracts have been prepared. Determination of biochemical indicators: red blood cell SOD activity was measured by pyrogallol autoxidation method [7]; serum M DA content was measured by TBA coloration method [8]; DNA and RNA extracts were measured by diphenylamine coloration method and orcinol respectively The content of DNA and RNA in each tissue was measured by colorimetry (5).
Protective effect of GCs (Total Glycoside of Cistanche) on radiation protection
1.3 The anti-radiation effect of GCs (Total Glycoside of Cistanche)
120 NIH mice weighing (21.0 ± 2.2) g, both male and female, were randomly divided into 4 groups: normal group, NS 20ml kg -1 day-1 for 18 days; the remaining 90 mice received 60Co gamma rays once Whole-body irradiation, the exposure dose is 6 Gy, and then divided into injury groups: NS 20ml·kg -1·day-1 immediately after irradiation; GCs (Total Glycoside of Cistanche) treatment group: 125 and 250mg·kg respectively after irradiation -1·d-1, 18 days each, 2 hours after the last administration of each group, the SOD activity in red blood cells, liver MDA content, DNA and RNA content in liver and spleen tissues, and liver and spleen were determined according to the aforementioned experimental methods. Observation of tissue morphology. All data are expressed as x ±s, and t-test processing is performed between groups.
2 Results
2.1 Anti-lipid peroxidation effect of GCs (Total Glycoside of Cistanche)
2.1.1 The effect of GCs (Total Glycoside of Cistanche) on SOD activity in erythrocytes and serum MDA content in normal mice: As can be seen from Table 1, GCs (Total Glycoside of Cistanche) 62.5 mg·kg -1 group is compared with the control group There was no difference (P> 0.05). GCs (Total Glycoside of Cistanche) 125, 250mg·kg -1 group can significantly increase the SOD activity in red blood cells (P <0.01) and reduce the serum MDA content (P <0.05).
Table 1: The effect of GCs (Total Glycoside of Cistanche) on SOD activity in erythrocytes and serum M DA content in normal mice
2.1.2 The effect of GCs (Total Glycoside of Cistanche) on the DNA content of various organs in normal mice: As can be seen from Table 2, compared with the control group, there was no significant difference in the DNA content of the heart and brain in each administration group (P>0.05) ). GCs (Total Glycoside of Cistanche) 62.5mg·kg -1 group had no effect on the DNA content of liver and kidney; GCs (Total Glycoside of Cistanche) 125mg·kg -1 group and 250mg·kg -1 group had liver DNA content compared with the control group increased by 9.6% (P <0.05), 14.2% (P <0.01); the content of DNA in the kidney increased by 15.4%, 16.0%, respectively, compared with the control group (P <0.01).
Table 2 Effects of GCs (Total Glycoside of Cistanche) on the DNA content of various organs in normal mice
2.1.3 The effect of GCs (Total Glycoside of Cistanche) on the RNA content of various organs in normal mice: As can be seen from Table 3, compared with the control group, there was no significant difference in the RNA content of the heart and brain in each administration group (P>0.05) ). The GCs (Total Glycoside of Cistanche) 62.5 mg·kg -1 group had no effect on the RNA content of liver and kidney; the liver RNA content of the GCs (Total Glycoside of Cistanche) 125 and 250 mg·kg -1 group increased by 18.1% and 26.2 respectively compared with the control group. % (All P<0.01); compared with the control group, the RNA content of the kidney increased by 20.9% and 25.4% respectively (all P<0.01).
Table 3 Effects of GCs (Total Glycoside of Cistanche) on the RNA content of various organs in normal mice
2.2 Anti-radiation effect of GCs (Total Glycoside of Cistanche)
2.2.1 The effect of GCs (Total Glycoside of Cistanche) on SOD activity in erythrocytes and M DA content in liver of irradiated mice: As can be seen from Table 4, the SOD activity in erythrocytes in the injured group was 46.8% lower than that in the normal group (P <0.01), Compared with the injury group, the GCs (Total Glycoside of Cistanche) 125 and 250 mg·kg -1 treatment group increased by 52.1% and 52.4% respectively (P<0.01). Compared with the normal group, the liver MDA content of the injury group was significantly increased by 24.3% (P <0.01), and the liver MDA content of each treatment group was significantly lower than that of the injury group by 18.4%, 17.9% (P<0.01), slightly higher than that of the normal group (P> 0.05).
2.2.2 The effect of GCs (Total Glycoside of Cistanche) on the liver DNA and RNA content of irradiated mice: The liver DNA and RNA content of the injured group was lower than that of the normal group, but there was no significant difference (P>0.05); the liver DNA of the treatment group Although the RNA content increased slightly compared with the injury group, the difference was not significant (P>0.05).
Table 4 Effect of GCs (Total Glycoside of Cistanche) on SOD activity in red blood cells and MDA content in liver of irradiated mice
2.2.3 The effect of GCs (Total Glycoside of Cistanche) on the DNA and RNA content of the spleen of irradiated mice: Table 5 shows that the DNA content of the spleen of the injured group was lower than that of the normal group by 15.5% (P <0.01), and each treatment group was more damaged The group increased by 14.9% and 15.2% respectively (P<0.01), which were close to the normal group (P>0.05). The RNA content of the spleen in the injured group was 18.6% lower than that of the normal group (P <0.05). Compared with the injured group, each treatment group increased by 15.0% and 20.3% (P <0.01), which were close to the normal group (P> 0.05).
Table 5 Effects of GCs (Total Glycoside of Cistanche) on DNA and RNA content of spleen of irradiated mice
2.2.4 The effect of GCs (Total Glycoside of Cistanche) on the morphology of the spleen of irradiated mice: Figure 1 below shows that in the injured group and the normal group, the nucleus staining in the splenic white pulp and spleen corpuscles is lighter, the DNA content is reduced, the cytoplasm staining is lighter, and the RNA content is significantly lower. Compared with the GCs (Total Glycoside of Cistanche) 125mg·kg -1 group and the injured group, the overall structure of the spleen was close to normal. The nucleus of the splenic corpuscle was stained with deep purple, the DNA content was significantly increased, the cytoplasm was stained with deep red, and the RNA content was significantly increased.
Figure 1 The effect of GCs (Total Glycoside of Cistanche) on the morphology of the spleen of irradiated mice
2.2.5 The effect of GCs (Total Glycoside of Cistanche) on the morphology of the liver tissues of irradiated mice: Figure 2 shows that the injury group has lighter staining of hepatocyte nuclei around the hepatic lobule portal area than the normal group, the DNA content is significantly reduced, and the cytoplasm is stained The liver cells become lighter, and the RNA particles of liver cells are significantly reduced. The GCs (Total Glycoside of Cistanche) 125mg·kg -1 treatment group is darker purple in the hepatocyte nucleus around the liver lobular portal area of the injury group, and the DNA content is significantly increased, and the cytoplasm is stained dark red, RNA The content has increased significantly.
3 Discussion
The SOD activity of human erythrocytes decreases significantly with increasing age and serum peroxides: The content has gradually increased [9]. In this paper, it was observed that the GCs (Total Glycoside of Cistanche) group significantly enhanced SOD activity in mouse red blood cells, decreased serum MDA content, and increased liver and kidney tissue Band RNA content, indicating that GCs (Total Glycoside of Cistanche) can supplement SOD consumption Or reduce the decrease in SOD activity, thereby enhancing the ability to scavenge endogenous free radicals, slowing down the rate of lipid peroxidation in the body, enhancing the antioxidant capacity, and reducing the damage of free radicals to nucleic acid and other biological macromolecules, prompting the protective effect of CGS (Total Glycoside of Cistanche) on nucleic acids is related to its anti-lipid peroxidation effect. It also suggests that GCs (Total Glycoside of Cistanche) may have the effect of promoting nucleic acid synthesis and slowing down the decomposition and destruction of nucleic acids, and further promote protein synthesis, thereby helping For the production of immune substances, accelerate the repair and regeneration of tissue cells, and improve the body's defense capabilities [6], its mechanism of action needs to be studied in depth. It has been reported in the literature that enhancing the immune function of the body can delay premature aging, and the nature of aging is closely related to nucleic acid metabolism[10]. Therefore, combined with the anti-lipid peroxidation and immune enhancement effects of GCs (Total Glycoside of Cistanche), it suggests that GCs (Total Glycoside of Cistanche) have anti-lipid peroxidation and immune enhancement effects. Total Glycoside of Cistanche has anti-aging effects.
Figure 2 The effect of GCs (Total Glycoside of Cistanche) on the liver tissue morphology of irradiated mice
After the body is irradiated with 60Co, a large number of toxic oxygen free radicals can be produced, causing a strong lipid peroxidation reaction, destroying nucleic acids, proteins, enzymes and other biological macromolecules, and causing cell and tissue damage [11]. It can be seen that the damaging effect of radiation is closely related to lipid peroxidation. The GCs (Total Glycoside of Cistanche) treatment group in this experiment can significantly promote the recovery of SOD activity in the red blood cells of the irradiated mice and the recovery of spleen nucleic acid content, and inhibit liver lipid peroxidation. Formation. It shows that the radiation protection effect of GCs (Total Glycoside of Cistanche) is important to prevent free radical damage. Histomorphology also shows that GCs (Total Glycoside of Cistanche) can restore the structure of liver and spleen damaged after irradiation, and can promote the recovery of nucleic acids, suggesting that the radioprotective effect of GCs (Total Glycoside of Cistanche) is also anti-lipid Peroxidation is related, and its mechanism needs to be further explored.
4 References
Gao Haiqian. Research on Chinese Patent Medicine, 1984;11(11):34
Sato Tiao et al. Foreign Medicine, Chinese Medicine and Chinese Medicine Section, 1987; 9(6): 33
Li Qiaoru et al. Shanghai Journal of Traditional Chinese Medicine, 1990; (11): 22
Yu Nancai et al. Chinese Journal of Traditional Chinese Medicine, 1990; 15(6): 22
Zhang Longxiang et al. Beijing: People's Education Press, 1984: 217
Ding Kexiang et al. Journal of Gerontology, 1987; 7(2): 42
Zhou G L et al. Biochemi Biophys, 1986; 4:71
Chen Shunzhong et al. J C lin Lab Tech, 1984; 2(4): 7
Wu Li et al. Journal of Gerontology, 1990; 10(6):353
Liu Junda. Journal of Gerontology, 1993; 13(6): 375
Kergonon J F et al. Biochimi, 1981;63:555
