| Purpose:Under the guidance of the theory of "preventing disease",through the establishment of BALB / c mouse model of influenza virus(IV)infection under haze conditions,to explore the prevention and treatment effect of volatile oil of Ganlu Xiaodu Micropill(GXM)on influenza,with a view to wearing sachets provide objective basis for influenza prevention;study the mechanism of GXM volatile oil intervention in mucosal immunity of mice infected with IV under haze conditions at the level of histology,molecular biology,etc.,explore its anti-IV infection target,for clinical application and development of traditional Chinese medicine preparations provide experimental basis.Material and method:72 SPF grade BALB / c mice were randomly divided into normal control group,model control group,preventive control group,and experimental observation group,with 18 mice in each group.4 groups of adaptive feeding,prevent the control group from nasal administration of volatile oil 7 days in advance,and start modeling on the 8th day,in addition to the normal control group,put each group in an artificial climate incubator for 2 hours,take out PM2.5the suspension was instilled into the nasal cavity once a day for 7 consecutive days,and was lightly anesthetized with ether on the 8th day of modeling.The FM1 strain of A type IV(0.1ml / piece)was inoculated through the nasal cavity.After that,the normal control group,the model observation group and the prevention control group were instilled with 0.9% saline in the same amount through the nose.The experimental observation group was administered GXM nasally once a day for 10 consecutive days.The mice in each group were collected on the 3rd,7th and 10 th days after exposure.Combined with the appearance of lung tissue,the results of lung tissue pathological light microscope and electron microscope,the degree of pathological inflammation of mouse lung tissue was scored and the mouse lung index was calculated.The gene content of p Ig R in mouse lung tissue was determined by q PCR method;ELISA method The expression levels of IL-6 and IL-17 in mouse serum were detected,and the expression levels of s Ig A in Bronchoalveolar lavage fluid(BALF)and Nasopharynxlavage fluid(NALF)were detected by the same method.Results:1.After PM2.5 combined with IV infection,in addition to the normal control group,the body weight of the remaining groups of mice after exposure was accompanied by different degrees of weight loss.The weight loss of the model control group mice was significant(P <0.001).The day began to decrease and reached the lowest value on the 7th day,which proved that the modeling was successful;compared with the model control group,the average weight change of the mice in the prevention control group and the experimental observation group was significantly different(P <0.05).Compared with the experimental observation group,there was no significant difference in weight change(P> 0.05).2.Visually observing the degree of lung lesions in mice after 7 days of co-infection,the lung tissues of normal control mice were pale pink,normal in size,no volume increase,no exudation,hyperemia,good elasticity,and soft texture without substantial changes.Lung tissues of model control mice showed obvious hyperemia and edema,dark red color,accompanied by dark red petechiae,increased volume,hard quality,and poor elasticity.In the control group,the lung tissue of the control group was smooth,light red,and had mild edema,no congestion,soft quality,and good elasticity.In the experimental observation group,mice with mild blood stasis and edema did not increase significantly after 7 days of IV infection,and were soft and poor in elasticity.3.Under the light microscope,the normal control group showed that the alveoli and alveolar septa of mice were intact,the size of alveoli was the same,the alveolar wall was thin,the interval was not widened,and there was no edema and congestion in the surrounding blood vessels.The alveolar,alveolar tube,alveolar sac and other structures of the model control group disappeared,the alveolar interval was significantly thickened,and a large number of inflammatory cell infiltrations were seen,and the blood vessels and bronchi were obviously around.The morphological structure of the alveoli and alveolar sacs in the control group of mice is relatively complete,with a small amount of inflammatory cell infiltration,accompanied by a small amount of edema and congestion,which is smaller and less severe than the model control group.In the experimental observation group,a small amount ofinflammatory cells infiltrated in the alveoli,and the alveolar interval thickened.Compared with the model control group,the edema and congestion were milder.4.The lung tissue of normal control mice has a complete cell structure and tight intercellular connections.The mitochondria,endoplasmic reticulum,Golgi complex,lamellar bodies and other organelles are abundant,and the nuclear membrane is clearly visible.In the model control group,the ultrastructure of the lung tissue of the mice was damaged,which showed that the nuclear membrane and the plasma membrane were blurred,the cilia in the alveolar cavity thickened and shed,the nuclear chromatin aggregated thick,the interstitial collagen fibers were accumulated in large numbers,and the arrangement was criss-cross and cells Necrosis,reduction of organelles,reduction or disappearance of mitochondrial crest,vacuole-like lesions in the endoplasmic reticulum,and a large number of virus particles in the cytoplasm.The number of type ? alveolar cells in the control group was increased,the morphology was basically normal,the microvilli were abundant,the alveolar septum thickening was not obvious,and a small amount of collagen fibers was seen in the interstitial.In the experimental observation group,the number of type?cells increased,lamellar bodies increased,fibroblasts and collagen fibers increased,but significantly less than the model control group,with swollen blood vessels and rich microvilli.5.Compared with the normal control group,the lung index and pathological score of mice in each infected group were increased at each time point(P<0.05).The model control group had higher lung index and pathological score at each time point after infection than the other groups,the difference was statistically significant(P<0.05),and reached the peak on the 7th day.After drug intervention,compared with the model control group,the lung index and pathological score of the control group and experimental observation group were reduced at each time point,and the difference was statistically significant(P<0.05);the lung index and pathological score of mice in the experimental observation group It was higher than the prevention control group on the 3rd day after infection,the difference was statistically significant(P<0.05);the lung index and pathological score of the experimental observation group on the 7th and 10 th days after infection were not statistically significant compared with the prevention control group P>0.05).6.After PM2.5 + IV co-infection,the expression of s Ig A in BALF and NALF of mice ineach group decreased at each time point,and the difference was statistically significant compared with the normal control group(P<0.05).After drug intervention,compared with the model control group,the levels of s Ig A in BALF and NALF in the prevention control group and experimental observation group were increased at each time point,and the difference was statistically significant(P<0.05).Compared with the experimental observation group,on the third day after infection,there was a statistically significant difference in the expression of s Ig A in BALF and NALF in the control group(P<0.05),and the expression of s Ig A in NALF in the two groups was the highest on day 3.On days 7 and 10,the expression of s Ig A in BALF and NALF in the experimental observation group was not statistically different from that in the control group(P>0.05).The expression of s Ig A in BALF was the highest on day10.7.Compared with the normal group,the serum levels of IL-6 and IL-17 in the mice at each time point were all increased and reached the peak on the third day,with statistically significant differences(P<0.05).After drug intervention,compared with the model group,both the nasal drip group and the prevention group decreased,and the difference was statistically significant(P<0.05).On the 3rd day,the expression of the prevention group was lower than that of the nasal drip group,and the difference was statistically significant(P<0.05).On the7 th and 10 th days,the expression of the nasal drip group decreased,but the difference was not statistically significant(P< 0.05).8.After PM2.5 + IV co-infection,the expression of p Ig R in mice decreased at each time point(P<0.05).After drug intervention,compared with the model control group,the expression of the prevention control group and the experimental observation group increased significantly on the third day(P<0.05);the expressions of the prevention control group and the experimental observation group continued to increase on the 7th and 10 th days.Statistical significance(P<0.05);compared with the control group,the expression of the experimental observation group was significantly lower on the third day after infection(P<0.05),and there was no significant difference between the two groups on the 7th and 10 th days(P> 0.05).Conclusion:1.IV infection has the characteristic of "wet toxicity" in the haze environment.In the earlystage of infection,mucosal tissue damage,inflammatory cell infiltration,tissue congestion and edema,and mucosal immune dysfunction are the key links of lung degeneration and IV infection.2.GXM volatile oil down-regulates the expression of IL-6,IL-17 and other inflammatory factors and up-regulates the expression of p Ig R,promotes the synthesis and secretion of s Ig A,regulates the mucosal immune function of mice,achieves its heat-clearing and detoxification,aromatization and humidity prevention and treatment of IV infection in the haze environment. |
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