Study On The Anti-fatigue Effect And Mechanism Of Dendrobium Huoshanense By Nourishing Yin And Tonifying Deficiency

BackgroundDendrobium huoshanense is a perennial herb of Dendrobium in Orchidaceae,which is one of the traditional Yin tonic herbs.It has the characteristics of long-term chewing with no slag,solid body and best efficacy.Dendrobium huoshanense is the best kind of medicinal sarcophagus with the reputation of "the first of Chinese immortal herbs".However,due to its narrow geographical distribution,natural breeding difficulties,and excessive mining and ecological environment damage,the wild Dendrobium huoshanense resources are extremely scarce,leading to the study of its pharmacological effects has not yet fully developed,as a result,it has not been explicitly included in The Chinese Pharmacopoeia,and this situation is not conducive to the rational application and deep development of Dendrobium huoshanense.With the growing maturity of Dendrobium huoshanensis artificial cultivation technology and the increase of its annual yield,it is necessary and practical to study its pharmacological effects in depth.The medical Dendrobium plants have good tonic effect.According to the current studies,Dendrobium huoshanense has the effects of anti-aging,treating pneumonia,blood lipid lowering,liver protection,immunity,anti-oxidation,blood sugar lowering,etc,but the the anti-fatigue effect of Dendrobium huoshanense has not been reported.In addition,there are few studies to explore its efficacy at the basic of its medicinal properties,and the material basis and mechanism of pharmacological action are still unclear.Therefore,with the guidance of TCM theory,based on the historical application of Huoshan Dendrobium,this study uses modern pharmacological research methods,,to verify the cold and hot medicinal properties of Dendrobium huoshanense,and to explore the deficiency-invigorating effect based on the correlation of sexual effects.ObjectivesAt basic of knowing the application history and characteristics of Dendrobium huoshanense,Dendrobium candidum and Liuwei Dihuang pill were used as control drugs,and the effects of the body state and cold and heat index of yin-deficiency mice were tested to verify the cold and cool medicinal properties of Dendrobium huoshanense in this study.Secondly,on the basis of the above research,the American ginseng with the effects of Invigorating Qi and nourishing yin,clearing heat and promoting body fluid,and the Ginseng with the effects of reinforcing vital energy were used as control drugs,the material basis and mechanism of the effect of nourishing yin and tonifying deficiency and anti-fatigue of Dendrobium huoshanense were studied by comparing the effects of raw powder,alcohol extract and water extract of Dendrobium huoshanense on the biological effects,material metabolism and energy metabolism of mice with kidney-yin deficiency and kidney-yang deficiency,which aimed to provide a reference for clarifying the pharmacological properties and efficacy of Dendrobium huoshanense and promoting its rational application.Methods1.Study on drug property verification and Anti-fatigue Effect of Dendrobium huoshanense.After 7 days of adaptive feeding,SPF healthy male Kunming mice were randomly divided into normal group,Yin-deficiency model group,Liuwei Dihuang pill group,Dendrobium officinale group,Dendrobium huoshanense A(0.33g/kg),B(1g/kg),C(2g/kg)and D(3.0g/kg)dose groups.The yin-deficiency animal model was prepared by continuous infusion of thyroxine(160mg/kg)for 14 days.1.1 Effects of Dendrobium huoshanense on cyclic nucleotides and thyroid hormones in mice with kidney-yin deficiency.On the 13th day of experiment,the temperature tendency of mice in each group was observed.On the 15th day of experiment,mice were sacrificed for cervical spondylolisthesis.Serum samples were taken from eyeballs for centrifugation.Serum cAMP and cGMP were detected by enzyme-linked immunosorbent assay.The ratio of cAMP to cGMP was calculated.The levels of T3 and T4 in serum were tested by radioimmunoassay.1.2 Effects of Dendrobium huoshanense on anti-fatigue,material metabolism and energy metabolism in mice with kidney-yin deficiency syndrome.The exhaustive swimming time was recorded on the 14th day of experiment,and then the levels of BLA,BUN,FFA,TG and CK in serum were tested by colorimetry.The contents of LDH?Na+-K+-ATPase and Ca2+-ATPase were measured by ultraviolet spectrophotometry.The contents of hepatic glycogen and myogen were detected by kit method.2.Pharmacological Validation and Anti-fatigue Effect of Different Extracts of Dendrobium huoshanense.180 male Kunming mice were randomly divided into 15 groups after 7-day adaptive feeding:normal group,Yin-deficiency model group,Yin-deficiency American ginseng group,Yin-deficiency Dendrobium huoshanense group,low and high dose groups of Yin-deficiency Dendrobium huoshanense water extract,low and high dose groups of Yin-deficiency Dendrobium huoshanense alcohol extract,Yang-deficiency model group,ginseng group,Yang-deficiency Dendrobium huoshanensis group,low and high dose groups of Yang-deficiency Dendrobium huoshanense water extract,low and high dose groups of Yang-deficiency Dendrobium huoshanense alcohol extract.Kidney-yin deficiency syndrome and kidney-yang deficiency syndrome animal models were established separately by continuous intragastric administration of thyroxine(160 mg/kg)and hydrocortisone(25 mg/kg)for 14 days.,2.1 Effects of different extracts of Dendrobium huoshanense on cyclic nucleotides and thyroid hormones in mice with kidney-yin deficiency and kidney-yang deficiency.On the 13th day of experiment,the temperature tendency of mice in each group was observed.On the 15th day of experiment,mice were sacrificed for cervical spondylolisthesis.Serum samples were taken from eyeballs for centrifugation.Serum cAMP and cGMP were detected by enzyme-linked immunosorbent assay.The ratio of cAMP to cGMP was calculated.The levels of T3 and T4 in serum were tested by radioimmunoassay.2.2 Effects of different extracts of Dendrobium huoshanense on anti-fatigue,material metabolism and energy metabolism of kidney-yin deficiency and kidney-yang deficiency.The exhaustive swimming time was recorded on the 14th day of experiment,and then the levels of BLA,BUN,FFA,TG and CK in serum were tested by colorimetry.The contents of LDH?Na+-K+-ATPase and Ca2+-ATPase were measured by ultraviolet spectrophotometry.The contents of hepatic glycogen and myogen were detected by kit method.Results1.Effects of Dendrobium huoshanense on cyclic nucleotides and thyroid hormones in mice with kidney-yin deficiency.Compared with the model group,the Yin-deficiency status of mice in the B,C and D dose groups of Dendrobium huoshanensis were improved,the body weight was significantly increased(P<0.001,P<0.01),the retention ratio of hot zone was significantly increased(P<0.001),the level of cAMP and the ratio of cAMP/cGMP were significantly decreased(P<0.01,P<0.001),and the contents of T3 and T4 in serum were significantly decreased(P<0.05,P<0.01,P<0.001).2.Effects of Dendrobium huoshanense on anti-fatigue,material metabolism and energy metabolism in mice with kidney-yin deficiency.Compared with the model group,the exhaustive swimming time of mice in the B,C and D dose groups of Dendrobium huoshanensis were significantly increased(P<0.001),the contents of hepatic glycogen and muscle glycogen were significantly increased(P<0.001,P<0.01),the levels of BLA,FFA,BUN and TG in serum were significantly decreased(P<0.05,P<0.01,P<0.001),the activity of LDH was significantly increased(P<0.001),the activities of CK,Na+-K+-ATPase and C a2+-ATPase were significantly decreased(P<0.001).3.Effects of different extracts of Dendrobium huoshanense on cyclic nucleotides and thyroid hormones in mice with kidney-yin deficiency and kidney-yang deficiency.Compared with the Yin-deficiency model group,the body weight of mice in the four groups of high and low dose groups of Dendrobium huoshanense water extract and alcohol extract groups of Dendrobium huoshanense were increased significantly(P<0.01,P<0.001,P<0.05),the retention ratio of hot zone were increased significantly(P<0.01,P<0.001),the levesl of cAMP in serum were decreased significantly(P<0.05,P<0.001,P<0.01),and the levels of T4 in serum were decreased significantly(P<0.05,P<0.001,P<0.01).The activity and mental state of mice in the high-dose water extract group and alcohol extract group of Dendrobium huoshanense were significantly improved,the value of cAMP/cGMP was significantly decreased(P<0.01,P<0.001),and the levels of T3 in serum were significantly decreased(P<0.001,P<0.05).Compared with the model group of Yang deficiency,the high-dose group of Dendrobium huoshanense water extract was significantly improved the autonomous activity and mental state of mice,the body weight was significantly increased(P<0.01),the retention ratio of hot zone was decreased(P<0.05),the cAMP/cGMP value was decreased(P<0.05),the contents of T3 and T4 were increased(P<0.001,P<0.05),and the content of T3 in low-dose group of Dendrobium huoshanense water extract was increased(P<0.05).4.Effects of different extracts of Dendrobium huoshanense on anti-fatigue,material metabolism and energy metabolism of mice with kidney-yin deficiency and kidney-yang deficiency.Compared with the Yin-deficiency model group,the contents of myoglobin in the four groups of high and low dose groups of Dendrobium huoshanense water extract and alcohol extract groups of Dendrobium huoshanense were increased significantly(P<0.05,P<0.001,P<0.01),the levels of FFA,BUN and TG were decreased significantly(P<0.01,P<0.001),the activities of LDH were increased(P<0.01,P<0.001),and the activities of CK and Na+-K+-ATPase were decreased(P<0.001,P<0.01).Compared with the model group,the exhaustive swimming time,hepatic glycogen content,serum BLA level and Ca2+-ATPase activity of mice in the high-dose water extract and alcohol extract of Dendrobium huoshanense groups were significantly increased(P<0.001),while the level of BLA in serum was decreased(P<0.01)and Ca2+-ATPase activities were significantly decreased(P<0.001).Compared with the model of Yang-deficiency,the exhaustive swimming time of mice in the high-dose group of Dendrobium huoshanense water extract was increased significantly(P<0.001),the contents of hepatic glycogen and muscle glycogen were increased significantly(P<0.001,P<0.01),the level of BLA was decreased(P<0.001),the levels of FFA,BUN and TG were increased significantly(P<0.05,P<0.001,P<0.01),the activities of CK,LDH and Na+-K+-ATPase were increased significantly(P<0.05,P<0.01,P<0.001).Conclusions1.Dendrobium huoshanense can improve Yin-deficiency in mice,regulate the disordered levels of cyclic nucleosides and thyroid hormones in Yin-deficiency.Its action trend is consistent with Dendrobium candidum and Liuwei Dihuang pills,showing cold medicinal properties.Its mechanism may be related to reducing the excessive energy consumption rate in Yin-deficiency.2.Dendrobium huoshanense can improve the body's exercise ability under Yin deficiency by regulating the content of energy metabolic enzymes,increasing the clearance rate of metabolites and increasing glycogen reserve.It shows the characteristics of nourishing yin and tonifying deficiency.The therapeutic effect of Dendrobium huoshanense is consistent with Dendrobium candidum and Liuwei Dihuang Pill.Its mechanism may be related to reducing the excessive energy consumption rate and increasing the removal rate of metabolites in Yin-deficiency.3.The water extract of Dendrobium huoshanense can regulate the body state,cyclic nucleotide level and thyroid hormone level of Yin-deficiency and Yang-deficiency syndrome,showing the characteristics of "conditional dominance,bidirectional applicability" of plain medicine.The alcohol extract of Dendrobium huoshanense can only regulate the body state,cyclic nucleotide level and thyroid hormone level of Yin deficiency syndrome,showing cold medicinal properties,which is the material basis of cold medicinal properties of Dendrobium huoshanense.4.The water extract of Dendrobium huoshanense can improve the anti-fatigue effect of Yin-deficiency and Yang-deficiency syndrome in mice,and has two-way regulation effect.Its mechanism is to reduce the excessive energy consumption under Yin-deficiency condition and improve the elimination of metabolites,increase the excessive slow energy consumption rate under Yang-deficiency condition and increase the removal rate of metabolites.It is the main material base of Dendrobium huoshanense for tonifying deficiency and anti-fatigue effect.Foundation.Alcohol extract of Dendrobium huoshanense can only enhance the exercise ability of fatigue mice with Yin deficiency syndrome.Its mechanism is to reduce excessive energy consumption under Yin deficiency condition and improve the elimination of metabolites,which accords with the characteristics of cold medicine.It is the main material basis of Dendrobium huoshanense for nourishing yin,clearing heat and resisting fatigue.