Effects Of Cistanche Deserticola On Acute Lung Injury in Rats With Sepsis

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

[Abstract] Objective: To study the effect of Cistanche deserticola on acute lung injury (ALI) in rats with sepsis. Methods: The model of sepsis was made by cecal ligation and puncture. The histopathological examinations were performed and the biological markers of ALI were measured. Results: Under a light microscope in the saline group, there was edema of lung interspace, a large number of erythrocytes and neutrophils, and plasma protein in the alveolar cavities. The lung wet /dry weight, the rate of neutrophils, and the protein content in the pulmonary alveolar lavage fluid pulmonary vascular permeability and pulmonary alveolar permeability and MDA, SOD, as well as MPO, were correspondingly significantly increased. In the group with Cistanche deserticola treatment, these indices of ALI were significantly lower than those in the group with saline.

Conclusion: Cistanche deserticola has great effects on ALI in rats with sepsis. The therapeutic mechanism is possibly through the inhibition of neutrophils and oxidative free radicals.

【Key words】Sepsis; Acute lung injury; Cistanche deserticola

Sepsis is a clinical emergency and critical illness, and seeking effective treatment drugs has always been a major problem in clinical and basic research. The traditional Chinese medicine pathogenesis of sepsis is the deficiency of righteousness and the exuberance of evil. The attacking therapy for the body's evil prosperity is one of the effective treatment measures. Cistanche is a traditional tonic Chinese medicine. Pharmacological studies have confirmed that it has protective functions such as enhancing the rate of cellular DNA synthesis and resisting oxygen-free radicals. The author experimentally observes the effect of Cistanche on lung problem damage in sepsis, in order to provide effective exploration for the treatment of sepsis.

Cistanche Echinacoside treat lung disease

1 Material and methods

1. 1 Animal Health 40 male 5D rats, weighing 200~250g, provided by the Experimental Animal Center of Huazhong University of Science and Technology, quality inspection number: TJLA—2008—1670 were randomly divided into 3 groups: 8 normal saline group, Ig/ There were 12 rats in the mL eggplant paste group and 12 in the 5g/mL Cistanche group. The experimental animals were given the corresponding drugs by routine gavage for 15 days, twice a day (the Cistanche deserticola extract was provided by the traditional Chinese medicine preparation room of this hospital), and the dosage of 200mg/kg was used in the experimental animals according to the method of literature U] 12 minutes after cecal ligation and perforation. Collection of specimens.

1.2 Rat lung histological examination. Take a small amount of the left lobe of the animal’s lung from each group, fix it with 10% formaldehyde, paraffin-embed the section, and HE stains, observes with an ordinary optical microscope, and on this basis, take the lung problem to make 1mm3 for 25 It was fixed in% glutaraldehyde solution, then fixed in 1% tetroxide solution, dehydrated with ethanol and acetone, embedded in Epon 81, prepared ultra-thin sections, stained with uranium-lead, and observed by transmission electron microscope.

1.3 Determination of wet/dry weight ratio of rat lung problem, cell count ratio in alveolar lavage fluid, and protein content Weight ratio. Another group of rats underwent alveolar lavage after thoracotomy, and the lavage fluid underwent cell count and cell classification under a microscope. Coomassie brilliant blue method is used to determine protein content and calculate lung permeability index (alveolar lavage fluid protein/plasma protein, LPI).

1.4 Changes in Pulmonary Vascular Permeability (PVP) in Rats According to the method introduced by Zhou et al. (2), after the administration, animals in each group were undergoing cecal ligation and perforation, and femoral vein injection of 50 mg/kg of lanthanum was used to open the chest and remove the lungs. Remove the surrounding problems, soak the lungs in the formamide solution (the amount of formamide is 20mg/100g body weight), incubated in a 45-50T incubator for 72h, wait until all the pigments in the problems are leached, take out the problems, centrifuge, and take the supernatant, Use an ultraviolet spectrophotometer to perform colorimetry at 620nm, and calculate the ethanol content according to the standard curve to determine the change of PVP.

1.5 Determination of malondialdehyde (MDA) in rat lung problem The thiobarbituric acid (TBA) method was used to determine MDA activity. Add 4mL 1.67mol/L sulfuric acid. 0.541% phosphonic acid to 50mL test specimen, mix well, centrifuge at 3000r/min for 10min, and process the precipitation once again as above, then add 1mL double-distilled water, 1mL TBA solution, 95% Anhydrous bath for 60 minutes, extraction with 5 mL n-butanol, tetra ethoxy propane as a standard, the final concentration of lnmol/L, fluorescence spectrophotometer at 532nm to measure the absorbance.

1.6 Determination of Superoxide Dismutase (SOD) in Rat Lung problems According to the method in literature [4], the SOD activity was determined by the Huangpuyin oxidase method (kit purchased from Nanjing Jiancheng Institute of Biological Engineering).

1.7 Determination of myeloperoxidase (MPO) in rat lung problem. Take lung problem to wash the residual blood as much as possible in isotonic saline, and place it in phosphate buffer containing 0.5% cetyl trimethyl bromide (PH6. 0) Make 5% of the homogenate, and operate according to the kit instructions.

1.8 Statistical methods Use SPSS statistical software, and measure data in (x±s) form, using t-test and X2 test.

Cistanche

2 results

2.1 Comparison of wet/dry weight ratio of lung problems, cell count ratio, and protein content in alveolar lavage fluid in each group are shown in Table 1. The lung wet/dry weight ratio in the saline group was significantly higher than that in the Cistanche group (P <0.05 or 0.01). The cell count ratio and protein content in the alveolar lavage fluid of the Cistanche group were significantly lower than those in the saline group (P <0.05 or 0.01). 0. 01), and with the increase of the dose concentration, it shows a significant decrease.

Table 1 Comparison of the wet/dry weight ratio of lung problem and the cell count ratio and protein content in the alveolar lavage fluid of rats in each group (X ± S)

Compared with the normal saline group, P <0. O1}}P<001. below is the same

2.2 Comparison of the morphological changes of the lung problems of the rats in each group. General observation: The lungs in the saline group were significantly enlarged and dark red, with varying degrees of congestion and edema on the surface, and a large number of pulmonary infarctions at the edge; the lungs in the Cistanche group were light red with a surface Mild congestion and edema, no obvious pulmonary infarction. Light microscope observation showed that in the saline group, there were alveolar and pulmonary interstitial congestion and edema, a large number of diffuse neutrophil infiltration in the alveolar cavity, some abscess formation, and varying degrees of atelectasis, necrosis, and compensatory emphysema. , Thrombus and similar transparent membrane formation. The above-mentioned lesions in the Cistanche group were significantly reduced. In the saline group, alveolar epithelial cells swelled, lamellar corpuscles emptied obviously, large vacuoles were formed, capillary endothelial cells were swollen, endothelial cells were continuously injured, the intercellular space widened, capillaries and alveolar basement membranes were loose and widened. Irregular thickening, uneven thickness. In the Cistanche group, alveolar epithelial cells were not swollen, lamellar corpuscle emptying was reduced, the number of vacuoles formed was small, the connection between capillary endothelial cells was close to normal, and the basement membrane was slightly loose and irregularly thickened. Compared with the Ig/niL cistanche group, the 5g/mL cistanche group had a smaller range of lung problem morphological changes, and the degree of lung injury was lighter.

2.3 Comparison of LPI and PVP of rats in each group is shown in Table 2. The LPI and PVP of the saline group were significantly higher than those of the Cistanche group (PV0.05 or 0.01). The LPI and PVP of the Cistanche group were lower, which had a significant dose-effect relationship.

2.4 The comparison of the activity of MDA.SOD.MPO in the lung problems of rats in each group is shown in Table 3. The MDA content in the lung problem of the saline group was significantly higher than that of the Cistanche group, while the SOD activity and MPO activity in the lung problem of the Cistanche group were significantly increased (F <0.05 or 0.01).

3 Discussion

Multiple organ failure is the root cause of the irreversible condition and high mortality rate in the late stage of sepsis (5). Protecting organ function is of great significance in reducing the high clinical mortality rate of sepsis. The lung is the first organ of multiple organ failure. 1/3 of patients with sepsis die from acute lung injury (ALI) and acute respiratory distress syndrome (ARDS). The main pathophysiological mechanism of ALI is the accumulation of a large number of neutrophils in the lung problem and It activates and adheres to vascular endothelial cells. At the same time, the concentration of reduced coenzyme II oxidase that transfers single electrons increases, causing respiratory bursts, generating a large amount of oxygen free radicals, and causing damage and function of cell membranes and membrane structures through strong oxidation. Obstacles⑹. Lipid peroxide reflects the change of oxygen free radicals in the problem and is directly related to the degree of lung injury (7). In the body's enzymatic and non-enzymatic anti-radical oxidation system, the activity of SOD maintains a good correlation with the degree of problem damage ⑺. Compared with the physiological saline group, the MDA and SOD activities of the Cistanche group showed statistically significant differences. Cistanche has the functions of enhancing immune function, anti-aging, and enhancing cell DNA synthesis rate. The experimental results showed that the lung coefficient, PLT, alveolar lavage fluid neutrophil ratio, and pulmonary vascular permeability of the Cistanche group were significantly decreased, MDA activity of lung problem decreased, SOD increased significantly, and MPO activity decreased. At the same time, morphological observation showed that the degree of lung problem damage was significantly reduced, the infiltration of neutrophils was reduced, and the 5g/mL Cistanche group had less lung injury than the Ig/mL Cistanche group, and there was a good dose-effect relationship. It suggests that Cistanche has a significant protective effect on ALI. The mechanism may be: (1) Inhibit the migration of inflammatory cells in the lungs, reduce the oxygen free radicals produced by neutrophils and other activated cells so that the biofilm is protected; (2) Promote nucleic acid and protein synthesis, and protect against lung injury Effect; (3) Enhance the immune function of phagocytes and reduce lung problem damage caused by endotoxin. The experimental results provide an experimental basis for the treatment of acute lung injury in clinical sepsis by strengthening the body and strengthening the deficiency.

references

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