Polysaccharides From Cistanche Can Significantly Delaying Aging Of Lung Function

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

Sun Yun Wang Dejun Liu Xiaomei Shi Le Zhang Hongquan

(Institute of Medicine, Yangzhou University, Yangzhou 225001)

Aging is a process in which tissues and organs of the body undergo functional and morphological degeneration and even aging in the course of life. The lung is directly connected to the external environment and accepts blood circulation from the whole body. It is most vulnerable to various harmful factors in the environment and in the body. It is one of the earliest organs in the body where aging occurs. The role of free radicals in the aging process has been confirmed by the research of many scholars. This experiment uses ozone to create an experimental aging model to observe the effects of polysaccharides from Cistanche on the hypoxia tolerance, antioxidant function, and ultrastructure of the lungs of aging mice. Influence. It is expected to lay the research foundation for polysaccharides from Cistanche in delaying pulmonary degenerative changes and health care of the elderly respiratory system.

The purpose of this study is to observe the degenerative changes in the ultrastructure of lung tissue cells in experimental aging mice and the influence of polysaccharides from Cistanche on it. Using ozone (Ozone, Ob) continuous inhalation for 4 weeks to induce lung aging model method, using Polysaccharide from Cistanche 50 and 100 grams to administer model mice for 6 weeks to detect hypoxia tolerance, lung, and serum superoxide in aging mice Dismutase (SOD) activity and malondialdehyde (MDA) content. The changes of lung tissue organelles and ultrastructure were observed under an electron microscope. Results The experimental aging mice caused by ozone significantly reduced the hypoxia tolerance; the lung and serum SOD decreased and MDA increased. Compared with the model group, Polysaccharide from Cistanche can significantly prolong the hypoxia tolerance time of aging mice (P<0.01), Improve the activity of SOD in the serum, reduce the content of MDA (P<0.01), and delay the degeneration of mitochondria and lamellar bodies in the lung cells of aging mice (P<0.01). Conclusion Polysaccharides from Cistanche can significantly delay the experiment Physiological degeneration and cell morphology of the lungs of sexually aging mice.

1 material

1.1 Animals

3 mon-age clean mice of ICR strain, 18~22g, ¥^ each half, provided by the Experimental Animal Center of Yangzhou University, certificate number: Su Dongzhi 98002^

1.2 Drugs and reagents

Polysaccharides from Cistanche: Provided by Nanjing Wild Plant Research Institute, prepared with distilled water to 2.5 g·LT, 5.0 g-L-1; SOD, MDA test kits are provided by Nanjing Juli Biomedical Engineering Research Institute.

1.3 Instrument

UV755B ultraviolet spectrophotometer, a product of Shanghai Analytical Instrument Factory; ozone generator, a product of Wuxi Medical Instrument Factory; H-300 transmission electron microscope, a product of Hitachi, Japan.

2 Method

2 1 Preparation of an experimental lung aging model with odor turn injury ICR mice were taken from the day and experimental design, and the mice in the model group and the treatment group were randomly grouped.

The experimental aging model was created for 4 weeks continuously. At the same time, the control group (take the normal mice purchased in the same batch) and the model group were given 20 ml-kg ^d'1 with normal saline, Polysaccharide from Cistanche treatment group I and II Polysaccharide from Cistanche 50, 100 mg-kg-1-d-' was given by intragastric administration, and the administration period was 6 weeks. After the last administration, various index tests were performed.

2.2 Hypoxia tolerance test method (2)

Each group of mice was put into a 250 ml grinding bottle containing 20 g of soda lime. Vaseline was sealed, and the time from putting the mice in the bottle to suffocation and death was recorded under normal temperature and pressure.

2.3 Lungs and blood

SOD^MDA content determination of each group of mice 1 h after the last administration, remove the eyeballs and collect blood to separate the serum; then the mice were sacrificed, the lung tissues were taken out, and saline was added to the glass homogenizer to prepare 100 g -f1 lung tissue homogenate, the SOD activity of lung and serum was determined by Huangsuyin oxidase method, and MDA content was determined by thiobarbituric acid condensation method.

2.4 Electron microscopy specimen preparation method (3)

The experiment is divided into control group, model group, and high-dose group. The modeling and administration methods are the same as above. After the last administration, 3 mice from each group are randomly selected, and they are put to death by necking, and the right sides are taken respectively. Lung apex tissue, 0.4 mmol, L-1 glutaraldehyde, and 0.04 mmol, double fixation with L'1 OfeO4, acetone gradient dehydration, Epon 812 embedding, lead-uranium electron staining, Hitachi H-300 transmission electron microscope with 10,000 to 20,000 magnification Observe the changes and structural changes of the main organelles under multiples.

2.5 The results of statistical processing are expressed in terms of tens of soil s, and the comparison of the two sample means uses the t-test.

3 Results

3.1 The effect of Polysaccharide from Cistanche on hypoxia tolerance in aging mice

One hour after the last administration of mice in each group, the hypoxia tolerance time was tested. The results showed that the time that aging mice could tolerate hypoxia was significantly shorter than that of normal mice. The time is significantly prolonged compared with the model group, but there is no significant difference compared with the control group, indicating that Polysaccharide from Cistanche can significantly improve the hypoxia tolerance of aging mice, see Table 1

3.2 The effect of Cistanche polysaccharide on the antioxidant function of aging mice

The blood (serum) and lung tissue (lung homogenate) of the mice in each group were collected after the last administration, and SOD and MDA were determined. Results The serum and lung SOD activity of aging mice decreased, and the MDA content increased. Compared with the control group, the difference was significant. Polysaccharides from Cistanche can increase the activity of serum and lung SOD in aging mice, and reduce the MDA content. Compared with the control group, there is no significant difference, and the difference with the aging model group is significant, see Tables 2 & 3.

3.3 The effect of Polysaccharide from Cistanche on the intracellular darkness of alveolar epithelial cells⑷

The manufacturing method of the electron microscope is the same as before. Morphometric analysis: Under the electron microscope, each specimen was observed in 10 fields of view to measure the organelles. The results showed that the mitochondria and lamellar bodies of the model group were reduced, the basement membrane was thickened, and the polysomes were increased. Compared with the control group, there were significant differences. Polysaccharides from Cistanche high-dose group improved it. Compared with the model group, the difference was significant, and it was basically close to the control group. See Tab 4.

3.4 Changes in the ultrastructure of lung epithelial cells in experimentally aging mice and the influence of Cistanche polysaccharides

Aging model mice can be seen under an electron microscope: alveolar type I epithelial cells are a single layer of flattened type with a thickness of about 0.2 pm. The nucleus is small and dense. There are a few mitochondria, Golgi complex, and scattered endoplasmic reticulum in the perinuclear cytoplasm. Relatively sparse, with fewer microfilaments and microtubules. The histological changes of the lungs of aging mice were observed under the electron microscope. The main changes were typed II alveolar epithelial cells. The microvilli on the free surface of the cells decreased or disappeared, the lamellar bodies were weakened, the lamellar structure was blurred, and a few cells had small lamellar cells. The body disappears, and it is a hollow or homogeneous amorphous strip-like substance with low electron density. Mitochondria and rough endoplasmic reticulum are reduced in the cell body. The basement membrane of alveolar epithelial cells is obviously thickened, and the proliferation of diaphragm cells and fibroblasts can be seen in the alveolar interstitium (Fig 1,2). Although the inner nuclear membrane and mitochondrial swelling were seen in the Cistanche polysaccharide group, the degree was significantly reduced compared with the model group (Fig 3.4). Observation under the electron microscope of the medication group showed that the type epithelial cells were flat, with a smooth surface, no nucleus, and some were about 0.1 thick. There were a few organelles in the perinucleus, and radium particles were visible. Type II cells have short villi on the surface, a large number of mitochondria, rough endoplasmic reticulum, and Golgi complex in the cytoplasm (Fig 5,6)

The ozone damage aging model is based on the free radical theory of aging. Oxygen-free radical damage plays an important role in the process of lung aging. The lungs are the only organ that communicates with the external environment. In a certain period of time, animals are continuously inhaled with ozone. The lungs are the first to be exposed to ozone and are most vulnerable to damage. After ozone enters the body, it quickly generates highly active superoxide anion free radicals. Excess free radicals exceed the body's scavenging ability, destroy and consume the free radical scavenging enzymes (such as SOD) in the body, and then cause the organs and even the body to age. In this experiment, the anti-hypoxia ability of aging mice caused by ozone decreased significantly, the activity of antioxidant substance SOD in serum and lung tissue decreased, and the lipid peroxidation metabolite MDA increased significantly. With the use of Rongrong Duojing 50,100 mg-kg1 continuous infusion of experimental aging mice, the aging mice have obvious hypoxia tolerance Increased, the activity of superoxide dismutase for scavenging free radicals is enhanced. The content of lipid peroxidation metabolites is reduced, indicating that polysaccharides from Cistanche can enhance the antioxidant function of aging mice.

Lamellar bodies in the cytoplasm of alveolar n-type epithelial cells are often used to measure the function of epithelial cells. The secretions of starvation lamellar bodies are called alveolar surfactants, which exist at the air-liquid interface of alveoli and can reduce The surface tension of the alveoli. The aging changes of the lamellar bodies can lead to the reduction of alveolar surface-active substances. The surface tension of the alveoli is relatively high, and the retraction force of the alveoli is increased, which results in the shrinkage of the alveoli. Electron microscopy observations revealed that the n-type alveolar epithelial cells of aging mice have changed significantly. It can be seen that the lamellar corpuscles are weakened, the structure is blurred, the mitochondria are swollen and reduced, the rough endoplasmic reticulum is also reduced, and the basement membrane is significantly thickened. Although similar structural changes can be seen after application of Polysaccharide from Cistanche, the degree is significantly reduced, indicating that Polysaccharide from Cistanche can significantly improve or delay the morphological degenerative changes in lung epithelial cell lamellar bodies and mitochondrial degeneration in aging mice. Most of the cell structure is basically intact, delaying the changes of lung ultrastructure. The results of this study suggest that polysaccharides from Cistanche can reduce the structural and functional changes caused by ozone-induced lung oxidative damage, delay lung aging, and provide experimental evidence for applied research.