| Sleep deprivation(SD)refers to the inability of sleep to meet normal physiological needs(generally less than 4 hours sleep for 24 hours)due to environmental changes(such as cold,high temperature,noise,etc)or other irresistible factors(such as high work pressure,combat status,etc),thus which can cause a series of changes in human physiology,psychology and behavior,such as the decline of motor ability,memory ability and immune ability.Hot and humid environment refers to the high temperature and humid environment where the temperature is above 29? and the relative humidity of air is over 60%.In the hot and humid environment,the human body bears both the heat transmitted from the outside and the heat generated inside the body,and the human body's evaporative and heat dissipation ability decline,the body accumulates too much heat,leading to organ damage and function decline,water and salt metabolism imbalance,internal homeostasis imbalance,resulting in physical and mental fatigue.At present,there are many reports about the effects of important organs by single condition of sleep deprivation or hot and humid environment.However,there is no report on the occurrence and development of fatigue and the corresponding protective measures against the damage of vital organs of the body under the dual conditions of the sleep deprivation and the hot and humid environment.Moreover,there is a lack of simple and effective drugs which can anti-fatigue and rapid recovery of fatigue.The research on the prevention and rapid recovery of military operations fatigue caused by sleep deprivation in hot and humid areas plays an important role in improving the combat effectiveness of troops,which is also an urgent problem to be solved in military operations fatigue.Therefore,we take Qi Huo decoction as our research object,which was an anti-fatigue formula previously studied,to explore its feasibility and effectiveness in solving the above problems.Qi Huo Decoction(QHD)is composed of Astragalus mongholicus,Rhodiola rosea,the fruit of Chinese wolfberry,Epimedium herb,Acanthopanax,Purslane and Gynostemma pentaphylla.It has the functions of invigorating qi and nourishing yin,nourishing essence and nourishing spirit,Clearing away heat and dampness,strengthening tendons and bones.Previous studies have shown that QHD has a certain role in general anti-fatigue.In this study,we observed the protective effect of QHD on important organs of fatigue rats under sleep deprivation and hot and humid environment,and explored its mechanism.so as to lay a foundation for QHD as a new drug selection preparation which preventing and rapidly restoring fatigue in military operations.?Research purposes?To observe the protective effect of QHD on liver,stomach,skeletal muscle of fatigue rats by sleep deprivation in hot and humid environment and ultrastructural observation,and to explore the feasibility of QHD as an anti-fatigue preparation.?Research methods?Forty male SD rats were divided into 4 groups by table of random number(10 each): blank control group(BC group),humid and hot control group(HC group),humid and hot sleep deprivation group(HS group)and humid and hot sleep deprived QHD group(HSQ group).Rats were allowed to live in the normal temperature and humidity chamber in BC group,and rats were allowed to live in the simulated hot and humid chamber in HC group,HS group and HSQ group.Rats were given distilled water to gavage for 15 days in BC group,HC group and HS group,and rats were given QHD to gavage for 15 days in HSQ group.Then rats were deprived of sleep for 72 hours in HS group and HSQ group.The liver,stomach,skeletal muscle damage of rats were detected,after 72 hours of experiment.The main detection indicators included:1.The serum concentrations of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)were detected.The liver tissues contents of hepatic glycogen,malondialdehyde(MDA)and reduced glutathione(GSH)were detected.Compared rats of each group hepatocytes expression of ?-galactosidase(?-gal).And the ultrastructure of rats hepatocytes was observed by transmission electron microscope.2.The pathological changes of gastric mucosa were observed by HE staining.The wall cell counts of the gastric mucosa were measured in the secretory phase by H+/K+-ATP enzyme fluorescence staining.3.The skeletal muscle tissues contents of creatine kinase(CK),lactate dehydrogenase(LDH),nitric oxide(NO),and inducible nitric oxide synthase(iNOS)were detected.And the ultrastructure of the gastrocnemius muscle tissue was observed by transmission electron microscopy.?Results?1.Observation of hepatoprotective effect:(1)Compared with the BC group,the data of the HC group,HS group and HSQ group about the serum concentrations of AST and ALT increased significantly(P<0.01).The data of the HS group and HSQ group about the serum concentrations of AST and ALT increased significantly than those in HC group(P<0.01).The serum concentrations of AST and ALT in HSQ group were significantly lower than those in HS group(P<0.01).The results showed that the high temperature and humidity environment could damage the liver function of experimental rats,while the continuous sleep deprivation aggravated the degree of injury,and QHD could effectively reduce the degree of injury.(2)Compared with BC group,the data of the HC group,HS group and HSQ group about liver tissues concentrations of hepatic glycogen and GSH decreased significantly(P<0.05),while liver tissues concentrations of MDA increased significantly(P<0.01).Compared with HC group,the data of the HS group and HSQ group about liver tissues concentrations of hepatic glycogen and GSH were significantly lower(P<0.01),while liver tissues concentrations of MDA increased significantly(P<0.01).Compared with HS group,the data of the HSQ group about liver tissues concentrations of hepatic glycogen and GSH were significantly higher(P<0.01),while liver tissues concentrations of MDA decreased significantly(P<0.01).These results showed that the high temperature and humidity environment could reduce the hepatic glycogen reserve of experimental rats,and cause the imbalance of oxidative stress in the body,leading to liver damage and slow recovery.Continuous sleep deprivation aggravated the process.QHD could increase hepatic glycogen reserve,maintain blood sugar stability,provide more energy,regulate oxidative stress balance,alleviate liver damage,and accelerate recovery,and maintain normal function of hepatocytes.(3)Microscopic observation showed that the hepatic tissue of had different degrees of inflammatory cell infiltration,hepatocyte swelling and necrosis on experimental rats of HC group,HS group and HSQ group.These results showed that high temperature and humidity environment and sleep deprivation resulted in inflammatory necrosis of liver tissue of experimental rats,and QHD could significantly reduce inflammatory necrosis.(4)Aging-related ?-gal expression was detected in hepatocytes of HC group,HS group and HSQ group.Compared with the BC group,the data of the HC group,HS group and HSQ group about the number of ?-gal positive cells increased significantly(P<0.01).Compared with the HC group,the data of the HS group and HSQ group about the number of ?-gal positive cells increased significantly(P<0.01).The number of ?-gal positive cells in HSQ group was lower than that in HS group(P<0.01).These results showed that high temperature and humidity environment could accelerate the aging process of hepatocytes,which could be accelerated by continuous sleep deprivation,while QHD could inhibit the aging of hepatocytes and reduce the damage of hepatocytes.(5)Transmission electron microscopy showed that the microvilli of hepatocytes decreased or even disappeared,and some hepatocytes vacuolated on rats of HC,HS and HSQ groups.The most obvious damage was rats of HS group,and the damage was alleviated to a certain extent on rats of HSQ group.These results showed that the ultrastructure of hepatocytes of experimental rats could be damaged in high temperature and humidity environment,and the degree of damage was aggravated by continuous sleep deprivation,and QHD could effectively reduce the damage.2.Observation of gastric protective effect:(1)HE staining showed that the gastric mucosa of rats were damaged to varying degrees in HC group,HS group and HSQ group.The gastric mucosa was scattered in a large number of punctate or linear erosion and congestion,with a small amount of blood scab on the surface,and inflammatory cell infiltration could be seen.The degree of gastric mucosal injury in HSQ group was significantly less than that in HS group.These results showed that high temperature and humidity environment could lead to gastric mucosal injury in experimental rats,and sleep deprivation could aggravate gastric mucosal injury,while QHD could significantly reduce the degree of gastric mucosal injury.(2)The rats gastric secretory phase parietal cells counts of HC group,HS group and HSQ group was significantly higher than that of the BC group(P<0.05).The rats gastric secretory phase parietal cells counts of HS group and HSQ group was significantly higher than that of the HC group(P<0.01).The rats gastric secretory phase parietal cells counts of HSQ group was significantly lower than that of the HS group(P<0.01).It was proved that gastric acid plays an important role in the formation of stress gastric mucosal damage which caused by sleep deprivation in hot and humid environment from the cytological level.QHD could reduce parietal cells into secretory phase,thus inhibited gastric acid secretion,thereby reduced the degree of gastric mucosal damage which caused by sleep deprivation in hot and humid environment.3.Observation of skeletal muscle protective effect :(1)Compared with BC group,the data of the HC group,HS group and HSQ group about gastrocnemius muscle tissues concentrations of CK and LDH increased significantly(P<0.01),while gastrocnemius muscle tissues concentrations of NO and iNOS decreased significantly(P<0.01).Compared with HC group,the data of the HS group and HSQ group aboutgastrocnemius muscle tissues concentrations of CK and LDH increased significantly(P<0.01),while gastrocnemius muscle tissues concentrations of NO and iNOS decreased significantly(P<0.01).There was no significant difference in the level of CK and LDH between HSQ group and HS group(P>0.05),The levels of NO and iNOS in HSQ group were significantly higher than those in HS group(P<0.01).These results showed that high temperature and humidity environment and sleep deprivation could cause energy metabolism disorder and free radical level imbalance of skeletal muscle.QHD could significantly improve the energy metabolism of muscle cells,regulate the balance of free radical levels,and enhance the antioxidant effect of the body.There was no significant difference in CK and LDH levels between HS group and HSQ group,which could be related to mild humid-heat environment,less damage to skeletal muscle cells caused by sleep deprivation and too few experimental samples.(2)Transmission electron microscopy showed: The gastrocnemius muscle fibrous sarcomere were clear,uniform in length,clear demarcation between light and dark zones,and regular in arrangement on rats of BC group and HC group.There were a series of changes in gastrocnemius muscle in HS group and HSQ group,such as slight dissolution of muscle fibers,unclear demarcation between light and dark zones,disordered arrangement,etc.Especially the ultrastructural changes were the most significant in HS group.These results showed that high temperature and humidity environment and sleep deprivation could lead to the destruction of skeletal muscle ultrastructure,while QHD could reduce the degree of skeletal muscle injury to a certain extent.?Conclusion and Significance?The above studies confirmed that:The liver,stomach and skeletal muscle of SD rats were damaged to some extent in the humid and hot environment.Continuous sleep deprivation could increase the severity of these injuries.Qi Huo Decoction could significantly improve the injury degree of liver,stomach and skeletal muscle of experimental rats.Qi Huo Decoction has protective effect on liver,stomach and skeletal muscle of fatigued rats induced by sleep deprivation in humid and hot environment.It could be used as a selection of traditional Chinese medicine preparations for fatigue prevention and rapid recovery. |
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