| PurposesWe established a intermittent hypoxic rats model,to simulate the sleep apnea hypopnea syndrom(OSAHS)patients of intermittent hypoxia state.We used the Xiaohanliqi Pill which is a exact clinical curative effect drug,and evaluated the drug for this model the influence of PI3K/Akt signal pathway in rats and the inhibition of lung inflammation,to explore the possible mechanism of Xiaohanliqi Pill in treating OSAHS.Methods:1.Our group manufacture intermittent hypoxic animal cabins2.Thirty male SD rats with SPF grade were randomly divided into blank group(NC),mild model group(IH-1),moderate model group(IH-2),mild Chinese medicine group(IH-3)and moderate Chinese medicine group(IH-4).After one week of adaptive feeding,except for the blank group,the other groups were placed in hypoxia chamber for 8 hours every day,and the rats were exposed to free food and activities for 5 weeks in total3.After 5 weeks of modeling,mild and moderate traditional Chinese medicine groups were given Xiaohanliqi Pill,and the other groups were given equal dose of normal saline.After 2 weeks of administration,the animals were sacrificed,and the right pulmonary alveolar lavage fluid(BALF)was extracted to detect the total number of white blood cells and cell classification.The left lung was fixed and embedded in paraffin section,and he staining was performed to observe the pathological changes of lung tissue in each group;the right upper lobe was taken for RT-qPCR detection,and the relative expression of PI3K and Akt mRNA in each group was compared;the average light of PI3K,Akt and PDK1 was measured by immunohistochemistry Density value.Results:1.Effects of IH on general conditions:Compared with the blank group,rats in each CIH group showed dim hair color,decreased activity,slow response,faster respiratory rate,and contradictory chest and abdomen movements.The above situation was alleviated in each traditional Chinese medicine CIH group after treatment.2.Effect of IH on the total number of white blood cells and cell classification count in BALF:The total number of white blood cells in the IH group increased,which was statistically significant compared with the blank group(P<0.05).There was a significant difference between the two different degrees of IH(P<0.01).Xiaohao Li Qi Granules significantly inhibited the growth of leukocytes in the 1H group(P<0.05),but the total number of leukocytes in the Chinese medicine hypoxia group was still higher than that in the blank group.Compared with the blank group,the proportion of neutrophils increased(P<0.05)and the proportion of monocytes decreased(P<0.05)in each 1H group.Compared with the IH model group,the proportion of neutrophils decreased and the proportion of monocytes increased(P<0.05)in the IH traditional Chinese medicine group,but the proportion of neutrophils was still higher than that in the blank group.The proportion of lymphocyte in each IH group was significantly increased(P<0.05).Comparison between groups showed that,except for IH-3 and IH-4,IH-2 and IH-3,pairwise comparison of other groups showed significant statistical differences(P<0.05)3.Pathological changes of bronchial and lung tissues:HE staining showed that inflammatory cell infiltration occurred to different degrees in lung tissues of all IH model groups,and the degree of inflammatory cell infiltration was significantly reduced in IH traditional Chinese medicine group compared with model group.4.Detection of protein expression level by immunohistochemistry:Compared with the blank group,PI3K,Akt protein expression in lung tissue of IH model group was significantly increased(P<0.05),compared with IH model group the AOD of IH traditional Chinese medicine group was significantly reduced(P<0.01).5.RT-qPCR detection:compared with the blank group,the expression of PI3K mRNA and Akt mRNA in lung tissue of IH model group was significantly enhanced(P<0.01),compared with the IH model group,the expression of PI3K mRNA and Akt mRNA in IH traditional Chinese medicine group was significantly decreased(P<0.01)but still higher than that in the blank group(P<0.05).Conclusion:Activation of PI3K/Akt signaling pathway is a key link in the initiation of pulmonary inflammation by intermittent hypoxia.By analyzing the paraffin section of lung tissue after he staining and the cell count and classification of BALF,it can be found that intermittent hypoxia can lead to inflammatory changes in lung tissue of experimental rats,and the inflammatory changes can be significantly inhibited by intragastric administration of xiaonao Liqi Pill.The results of IHC and RT qPCR showed that the expression of PI3K and Akt in lung tissue of intermittent hypoxia group was significantly increased,while the expression of PI3K and Akt in Chinese medicine group was significantly decreased.Xiaohanliqi Pill can reduce the infiltration of inflammatory cells and alleviate pulmonary inflammation by inhibiting the transmission of PI3k/Akt signal in cells. |
PDF Full Text Request