Desertliving Cistanche Anti- Aging Action：A Comparative study

Contact: Audrey Hu Whatsapp/hp: 0086 13880143964 Email: audrey.hu@wecistanche.com

XU Hui, WEI Xiao - dong, OU Qin Li Ying Xu Xiao - Xia, WANG Jing - tao ₉ TIAN Guo - Zhong
(Jiamusi University» Jiamusi 154007, China)Abstract

Objective: To investigate the age-related changes in anti-oxidation of rat liver mitochondria induced by D-gal and the anti-aging effects of desertliving cistanche and its water extract, ethanol solution, and polysaccharide components at the level of mitochondria. Methods We used the aging rat induced by D—gal as a research sample. We measured the Ca2+-ATPase activity in rat liver and determined the MDA content, membrane fluidity, and PLA2 activity in rat liver mitochondria in an attempt to study the effects of different components of desert living cistanche on the above—mentioned indexes. Results: The MDA content and PLA2 activity were markedly decreased, and the Ca2+-ATPase activity and membrane fluidity were obviously increased bygiving the water extract, ethanol solution, and polysaccharide components to the desert living cistanche groups (P<2 0. 05 <C 0. 01). In the water extract group» the water extract was able to cause the above indexes up to the level of the young group but the effects of ethanol solution and polysaccharide components were relatively inferior. Conclusion: Desertliving cistanche different components can postpone aging by raising the liver anti-oxidation function, reducing the damage by the peroxidation of the liver mitochondria, with the effect of desert living cistanche water extract better than that of other components.

Keywords： desertliving cistanche； anti-aging； mitochondrion

Cistanche deserticola has many effects, click here to know more

The traditional Chinese medicine Cistanche is the dry fleshy stem of Cistanche, which has the functions of nourishing kidney yang, nourishing essence and blood, and moisturizing the intestines. It has a certain pharmacological effect on various system functions of the body. It is a traditional Chinese medicine for tonic. Experiments have confirmed that it has anti-aging effects, but the comparison of the anti-aging effects of different components of Cistanche and the research on the anti-oxidative damage of mitochondria at the level of mitochondria and the mechanism of improving mitochondrial function and mechanism have not been reported at home or abroad. This experiment studies the anti-aging effects of the different components of Cistanche and provides the theoretical and experimental basis for studying the anti-aging mechanism of the aging mechanism and Chinese herbal medicine components.

Anti-aging effects of cistanche

1 Experimental material

1.1 Experimental animals

50 pure Wistar rats, bodyweight 180～210g, 3～4 months old, half male and half male, purchased from the Experimental Animal Center of Jiamusi University

1.2 Experimental drugs

Cistanche was purchased from Jiamusi Medicinal Material Company. Take 400g of crude drug, soak it in water for 30 minutes, and extract twice by ultrasound. After every 30 minutes, after suction filtration, the filtrate is combined and concentrated to equivalent to 0.5g of crude drug per milliliter, and then stored for use; then 400g of the crude drug is prepared according to the above method to obtain an aqueous extract. Repeated precipitation by adding ethanol according to the literature method, suction filtration to separate the precipitate, washing the precipitate with absolute ethanol, acetone, ether, drying, and constant weight to obtain Cistanche deserticola polysaccharide, storage at 4C for later use; the ethanol-dissolved part was concentrated under reduced pressure to alcohol-free Taste, store at 4℃, make it to the required concentration before use.

1.3 Main reagents and instrument reagents:

DPH was purchased from Sigma, HEPES was purchased from Amreseo, and NADH was purchased from Amresco. MDA test kit and Ca-ATPase test kit were purchased from Nanjing Jiancheng Bioengineering Research Institute, and D-galactose was purchased from Shanghai Second Reagent Factory. Apparatus: Shimadzu RF-5000 fluorescence spectrophotometer, Sartorius PP-25 high-sensitivity PH meter, GL-21M high-speed freezing centrifuge, RE-52A rotary evaporator, SHB-Ⅲ circulating water multi-purpose vacuum pump.

Cistanche deserticola extract

2 Experimental method

2.1 Animal medicine and treatment

Fifty rats were randomly divided into a young control group and a D-galactose model group. The model was given to the Cistanche water extract group, the model was given to Cistanche cistanche alcohol-soluble groups, and the model was given to the Cistanche polysaccharide group, each group was 1oN. All rats are kept in separate cages in the same room. The indoor temperature is 16-22C, and the humidity is 50-65%. They eat and water normally every day. Natural light. The young control group was injected with saline subcutaneously every morning, the dose was equivalent to the dose of a gal in other administration groups, and was given warm boiled water at the same time. In the aging model group, D-gal was injected subcutaneously at a dose of 50 mg/kg every morning, and at the same time filled with warm boiled water. The model gives the Cistanche water extract group (referred to as the water extract group) subcutaneously injects D-gal at a dose of 50mg/kg every morning, and at the same time administers the cistanche water extract at a crude drug amount of 2g/kg. The model gave Cistanche alcohol-soluble group (abbreviated as an alcohol-soluble group) every day morning and afternoon at a dose of 50mg/kg subcutaneous injection of D-gal, at the same time the crude drug amount 2g/kg water extract of cistanche water extract alcohol-soluble components. The model gave the Cistanche polysaccharide group (referred to as the polysaccharide group) subcutaneous injection of D-gal at a dose of 50 mg/kg every morning. At the same time, administer Cistanche Polysaccharides at a rate of 2g/kg of the crude drug. After continuous administration of 6N, the animals were put to death. Take material.

2.2 Index measurement

Take 1g of the liver. Rinse with cold salt water, wipe dry and weigh. Add 9 times the volume of pre-cooled homogenization medium (pH 7.4, 0.01mol/L Tris-HC1, 0.0001mol/LEDTA-2Na, 0.01mol/L sucrose, 0.8 sodium chloride solution), electric The homogenizer fully grinds to obtain liver tissue homogenate, and the liver tissue homogenate is centrifuged at 3000r/rain for 10 minutes. Take the supernatant and freeze it for testing. Prepare liver homogenate according to the above method, take 10% liver tissue homogenate and centrifuge at 2000r/min for 10min, discard the precipitate, take the supernatant 10000r/rain, and centrifuge for 15rain in a high-speed refrigerated centrifuge, the precipitate is mitochondria, and the mitochondria are suspended in Medium. For index determination. Measure the Ca～ATPase activity of liver tissue and the content of malondialdehyde (MDA) in liver mitochondria according to the instructions of the test kit. The lipase A activity and liver mitochondrial membrane fluidity was measured according to the literature.

2.3 Statistical analysis

The experimental data are expressed as mean ± standard deviation (± s), and SPSS11.5 statistical software is used for statistical analysis.

Effects of cistanche: Anti-aging

3. Results

In the aging model group, the activity of Ca-ATPase in the liver and the fluidity of the liver mitochondrial membrane decreased, and the content of MDA and PLA activity of liver mitochondria increased. The different components of Cistanche can significantly increase the Ca-ATPase activity of the liver in aging model rats, the fluidity of the liver mitochondrial membrane, and significantly reduce the MDA content and PLA activity of liver mitochondria. The effect of Cistanche water extract is more obvious. See Table 1.

4. Discussion

In recent years. Research has found that mitochondrial oxidative damage and functional changes play an important role in the aging process of cells, tissues, and organs]. The free radical theory of aging has always been an important theory among the many aging theories, and the "free radical-mitochondrial damage" hypothesis about aging is currently generally accepted [5]. It further enriches the theory of free radicals. This hypothesis believes that mitochondria are the main oxygen-consuming parts of cells and the main sites for generating free radicals. Therefore, they are also the target parts of free radicals. Therefore, the mitochondria, an organelle, is important foraging and traditional Chinese medicine. The study of the anti-aging mechanisms is of great significance to reveal the essence of aging. Cistanche is a tonic Chinese medicine. Its anti-aging effect has been confirmed, but it is related to its anti-aging mechanism at the mitochondrial level and the effect of different components of Cistanche on the anti-aging effect of mitochondria. Research, there is no report at home and abroad. In this experiment, Cistanche water extract, alcohol-soluble components, and polysaccharides were used as application factors to explore the anti-aging mechanism of different components of cistanche at the mitochondrial level. Many experimental pieces of evidence show that D-galactose aging model rats have a series of changes similar to natural aging L6]. In aging, the SOD activity in the cell, especially the mitochondria, decreases. MDA content increased.]. The results of this experiment show that the content of malondialdehyde (MDA) in liver mitochondria of aging rats has increased significantly, which indicates that the reduction of antioxidant scavenging rate during aging reduces the body’s antioxidant capacity, which in turn makes the production and elimination of reactive oxygen species and other free radicals out of balance. As a result, reactive oxygen species accumulate in the body and damage cells. In addition, the damage to cells caused by calcium overload mainly has two aspects: ① uncoupling of mitochondrial oxidative phosphorylation and inhibition of cell respiration; ② activation of phospholipase c and A. , A. This experiment verified that the activity of Ca-ATPase in the liver tissue of aging rats decreased, and the liver mitochondria showed PLA. The increase in activity indicates that the decrease in Ca-ATPase activity may increase the concentration of cytosolic Ca, leading to an increase in mitochondrial PLAz activity. PLA: It is a key enzyme in many biochemical processes. It can hydrolyze its own phospholipids to increase the fragility of the mitochondrial membrane framework, increase the release of arachidonic acid, and generate a large number of free radicals, which leads to an increase in membrane permeability and a large influx of calcium ions. Activate PLA L8], thus creating a vicious circle. In addition, the increase in PLA activity can change the composition of membrane phospholipids and affect membrane fluidity. Maintaining proper membrane fluidity is essential for electron transport, proton transport, and oxidative phosphorylation. In this study, the fluorescence polarization method was used to find that the mitochondrial membrane fluidity of the liver of aging rats was significantly lower than that of young rats, further confirming that mitochondrial membrane fluidity changes during aging.

This experiment showed that the water extract of Cistanche deserticola, the alcohol-soluble components of the water extract of Cistanche deserticola, and the polysaccharides of Cistanche deserticola all improved the activity of liver Ca-ATPase and the fluidity of mitochondrial membrane to varying degrees, and reduced the content of MDA and PLA: activity of liver mitochondria. It proves that the different components of Cistanche have anti-aging effects. The mechanism may be due to the improvement of the antioxidant capacity of each administration group, which ensures the activity of Ca and even ATPase, so that the calcium ion concentration inside and outside the liver cells maintains a normal level, avoiding the increase of mitochondrial PLA activity due to intracellular calcium overload. High thereby reduces the influence of various factors that reduce the fluidity of the mitochondrial membrane, keeps the fluidity of the mitochondrial membrane normal, and ensures the function of the mitochondria. The results also showed that the Cistanche aqueous extract can restore the indicators of the medication group to the level of the young group, and its effect on the aging model group is better than other components. The literature reported that Cistanche deserticola polysaccharide can increase lung tissue SOD activity and reduce MDA content [9]. Xue Dejun also proved that Cistanche deserticola polysaccharide can activate serum SoD activity and reduce cardiac muscle lipofuscin_]. This experiment shows that Cistanche deserticola polysaccharide has anti-aging effects. This may be because the cistanche deserticola polysaccharide can improve immune function, activate SOD to directly or indirectly scavenge free radicals, reduce the damage of free radicals to mitochondria and the whole cell, and thus play an anti-aging effect. The alcohol-soluble components of Cistanche water extract can improve immunity and androgenic hormone-like effects [1, this experiment found that it also has anti-aging effects, which may be due to the anti-aging effects of the flavonoids and glycosides contained in it. The anti-aging effect of Cistanche water extract is better than other ingredients, indicating that the anti-aging effect of Cistanche water extract is the result of the synergistic effect of its two components. At the same time, Cistanche contains inorganic elements such as Fe, Zn, Cu, which are all necessary for the body Trace elements, Fe is the active center or activator of enzymes, proteins, hormones, and vitamins; Zn can maintain various enzyme activities; Cu is the main component of various metal enzymes and is an effective catalyst for the body's redox system. It can be seen that the anti-aging effect of Cistanche depends on its organic active components (polysaccharides, etc.) and inorganic elements to work together.

Benefit of cistanche extract: anti-aging

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