| Depression is a kind of psychosomatic disease characterized by significant and lasting depression,lack of interest,slow thinking and cognitive function.It has four characteristics:high morbidity,high disability rate,high suicide rate and high recurrence rate.It seriously endangers people’s physical and mental health and has become the focus of common concern all over the world.The symptoms of depression are complex,not only with central mental and psychological symptoms,but also with peripheral somatic symptoms.Its pathological mechanism is quite complex,but the monoamine transmitter hypothesis is still the mainstream academic viewpoint.The application of antidepressants based on the monoamine transmitter hypothesis has brought about a historic change in the treatment of depression.It can effectively alleviate depressive psychiatric symptoms such as depression and loss of interest.However,there are also objective problems such as slow onset of drugs(2-4 weeks)and more residual somatic symptoms.Depression is a bottleneck in clinical treatment.Mitochondria are the "power plant" of eukaryotic cell life activities.Mitochondrial damage can cause cell dysfunction and lead to symptoms of multi-system disorders,which is very consistent with the depression manifestation of systemic multi-system dysfunction.In recent years,more and more studies have shown the relationship between mitochondria and depression.The clinical patients or animal models of depression have deficiencies in the function and structure of mitochondria in many parts of the body.The condition(behavior)and somatic symptoms of post-mitochondrial depression are also improved.Depression can be either the cause or the result of mitochondrial damage.Traditional Chinese medicine treatment of depression mostly starts with the liver and spleen,and the theory of liver and spleen co-governance is also a classic theory of traditional Chinese medicine for the treatment of depression by physicians of past dynasties.In recent years,many scholars have also found that liver depression and spleen deficiency syndrome are the most common symptoms of depression through a large sample of epidemiological research on traditional Chinese medicine syndromes of depression.The theory of "treating depression from the liver" is still dominant,but the psycho-psychological symptoms such as low mood,lack of interest and physical symptoms such as extreme fatigue,slow thinking and loose stool can be regarded as multi-system and low motility symptoms,which are highly consistent with the symptoms of functional damage of spleen in traditional Chinese medicine.Therefore,"Treating depression from spleen" has become another important direction of treatment for depression.Academician Wang Yongyan put forward a new method of treating depression from the spleen and resuscitating the spleen and relieving depression.The "resuscitating the spleen and relieving depression prescription" created by the research group based on this theory has also shown remarkable effect on depression in a large number of clinical practice and previous animal experiments.The role of the spleen as the "acquired basis" is highly consistent with the understanding that mitochondria is the "hub of human life activities".In animal models of spleen deficiency,the mitochondrial content of heart,liver,stomach,muscle and other cells decreases.Therefore,the mitochondrial mechanism may be the effective mechanism of treating depression from the spleen theory.SIRT1/PGC-1 alpha pathway is the key pathway of mitochondrial biosynthesis and respiration,and plays an important role in maintaining the ability of mitochondrial oxidative phosphorylation to synthesize ATP.Therefore,we hypothesize that the abnormal mitochondrial biosynthesis mediated by SIRT1/PGC-1a pathway may be the biological essence of the pathogenesis of liver depression and spleen deficiency in depression,or the mechanism of Xingpi Jieyu recipe in treating depression of liver depression and spleen deficiency.This study intends to observe whether there is abnormality of SIRT1/PGC-1a pathway in depression patients with liver depression and spleen deficiency through clinical trials,and further explore the clinical efficacy of Xingpi Jieyu Formula on depression patients with liver depression and spleen deficiency and whether it has regulatory effect on SIRT1/PGC-1a pathway;if Xingpi Jieyu Formula can regulate SIRT1/PGC-1a pathway,cell experiments will further clarify the awakening effect.Whether Pijieyu Recipe plays an anti-depressive(protective)role through SIRT1/PGC-1alpha pathway.Clinical Research:Study on the Biological Essence of Liver Depression and Spleen Deficiency Syndrome in Depression and the Effect Mechanism of Xingpi Jieyu Prescription Based on SIRT1/PGC-1a PathwayObjective:To explore the correlation between liver depression and spleen deficiency syndrome of depression and SIRT1/PGC-1alpha pathway,to explore the clinical efficacy of Xingpi Jieyu recipe and whether there is a regulatory effect on SIRT1/PGC-1alpha pathway.Methods:Firstly,through clinical trial 1,10 patients with calm health,depression of Liver-qi Depression and depression of Liver-qi Depression and spleen deficiency were included.The levels of SIRT1,PGC-1a,NRF-1,NRF-2 and TFAM in peripheral blood were detected by Western Blot method.Then a randomized,double-blind,placebo-controlled clinical trial was conducted.61 patients with depression due to liver depression and spleen deficiency were randomly divided into control group(30 cases)and experimental group(31 cases).The control group was given placebo(Xingpi Jieyu Granule Simulator),one dose a day;the experimental group was given Xingpi Jieyu Granule,one dose a day.Both groups were treated for 6 weeks.The patients were assessed by HAMD,HAMA and TCM syndrome factors scale of depression on the day of admission and 6 weeks after treatment.SDS and SAS were assessed on the day of admission and at the 2nd,4th and 6th weeks after admission.Blood samples were collected before and after treatment.ATP,ADP and AMP were detected by high performance liquid chromatography.SIRT1,PGC-1a,NRF-1,NRF-2,TFAM protein and RNA were detected by Western Blot and RT-PCR.Safety tests were performed before and after treatment,including ALT,AST,CK,BUN,blood routine,urine routine and ADR scale.Results:In terms of clinical efficacy indicators,The total scores of HAMD,anxiety/somatization,cognitive impairment,day-night change,block and despair in the control group were significantly lower than those before treatment(p<0.01 or P<0.05).The total scores of HAMD and anxiety/somatization in the experimental group were significantly lower than those before treatment(p<0.01 or P<0.05)."Sleep disorder","cognitive impairment","day and night change","blockade","despair","gastrointestinal symptoms","systemic symptoms" 2 single factors,HAMA total score and "somatic anxiety","mental anxiety" 2 factors,SDS total score,SAS total score,TCM syndrome "Qi deficiency","Qi stagnation" and "phlegm dampness" scores were significantly decreased(p)<0.01),the total effective rate of HAMD score was 93.55%(marked efficiency was 70.97%),which was significantly better than 66.67%(16.67%)of the control group.In addition,compared with the control group,the total scores of HAMD,"gastrointestinal symptoms","systemic symptoms","anxiety/somatization","sleep disorder","block","HAMA total score","somatic anxiety","Qi deficiency" and "Qi deficiency" of TCM syndromes in the experimental group were higher than those in the control group.The total scores of stagnation,phlegm-dampness,SDS and SAS in 2-6 weeks decreased significantly(p<0.01 or P<0.05).In terms of mechanism indexes,SIRT1 protein content in peripheral blood of depression patients with Liver-qi Depression was higher than that of healthy people(p<0.05),while SIRT1,PGC-1a and TFAM protein content in peripheral blood of depression patients with Liver-qi Depression and spleen deficiency were lower than those of healthy people(p<0.05).Compared with before treatment,the AMP content in peripheral blood increased after treatment in the control group(p<0.05);the ATP content in peripheral blood increased after treatment in the experimental group(p<0.05);the ATP content in peripheral blood increased after treatment in the experimental group compared with the control group(p<0.05),while the ADP and AMP content decreased after treatment(p<0.05).There was a significant difference(p>0.05).After treatment,the levels of SIRT1,PGC-1a,NRF-1,NRF-2 and TFAM protein in peripheral blood of the experimental group were significantly higher than those of the control group(p<0.01).RT-PCR results showed that there was no significant difference before and after treatment in the control group(p>0.05).After treatment,the levels of NRF-1,NRF-2 and TFAM in peripheral blood of the experimental group were significantly higher than those before treatment and after treatment in the control group(p>0.05).The contents were significantly increased(p<0.01).In terms of safety indicators,one patient in the control group had nausea,and two patients in the experimental group had diarrhea.They all recovered spontaneously in a short period of time(2-3 days).There were no cases of liver and kidney damage.Conclusion:Compared with healthy people and patients with depression of Liver-qi stagnation,patients with depression of liver-qi stagnation and spleen deficiency have low expression of SIRT1/PGC-1alpha pathway in peripheral blood.Compared with placebo,Xingpi Jieyu Formula can significantly improve depression and somatic symptoms in patients with depression,increase ATP content in peripheral blood,decrease ADP and AMP content,and up-regulate SIRT1/PGC-1alpha pathway expression.Pathway-mediated abnormal mitochondrial biosynthesis is one of the biological essence of depression due to liver depression and spleen deficiency.Basic Research:Regulatory Mechanism of Xingpi Jieyu Recipe on SIRT1/PGC-1a Pathway in PC 12 Cells Injured by CorticosteroneObjective:To explore whether Xingpi Jieyu Recipe has antagonistic effect on PC 12 cells injured by corticosterone,and to clarify whether SIRT1/PGC-1alpha signaling pathway plays a mediating role in the anti-corticosterone neurotoxicity of Xingpi Jieyu Recipe.Methods:Eight-week-old SPF Wistar male rats were given 27g(crude drug)/kg body weight by intragastric administration for 7 consecutive days.The serum containing Xingpi Jieyu Formula was prepared by taking blood from abdominal aorta one hour after the last administration.PC 12 cells in logarithmic phase were inoculated on 96-well plate.After 24 hours of adherence growth,the culture medium was replaced with Wistar rat serum containing 5%,10%and 20%of drug-containing serum,respectively,and then cultured for 24 hours.CCK-8 solution was tested.PC 12 cells were inoculated on 6-well plate and divided into blank control group,model group,model group+SIRT1 inhibitor EX527 group,Xingpi Jieyu recipe+SIRT1 inhibitor EX527 group,CCK-8 method was used to determine the cell survival rate,Western blot was used to detect the expression of SIRT1,PGC-1a,NRF-1,NRF-2,TFAM protein,and PGC-1a siRNA was transfected into PC 12 cells.The transfected cells were divided into blank control group,model group,model group+PGC-1 alpha silencing group,Xingpi Jieyu recipe+PGC-1alpha silencing group,Xingpi Jieyu recipe+PGC-1alpha silencing group.The cell survival rate was determined by CCK-8 method.The expression of SIRT1,PGC-1a,NRF-1,NRF-2 and TFAM protein was detected by Western blot.Results:Compared with the blank control group,the OD value of PC12 cells in the model group decreased significantly(p<0.01),the survival rate decreased,and the protein expressions of SIRT1,PGC-1a,NRF-1,NRF-2 and TFAM decreased significantly(p<0.05 or P<0.01);compared with the model group,the OD value of model+EX527 group and model+PGC-1a silent group decreased significantly(p<0.01),the cell survival rate decreased,and the OD value of traditional Chinese medicine group increased significantly.(p<0.01),cell viability increased,SIRT1,PGC-1a,NRF-1,NRF-2,TFAM protein expression increased significantly(p<0.05);compared with the traditional Chinese medicine group,OD value decreased significantly(p<0.01),cell viability decreased,SIRT1,PGC-1a,NRF-1,TFAM protein content decreased(P<0.05),OD value decreased significantly in the traditional Chinese medicine+PGC-1a silence group(p<0.01).The cell viability and the contents of PGC-1a,NRF-1 and TFAM decreased(P<0.05).Conclusion:The serum containing Xingpi Jieyu Formula exerts its reversal effect on CORT-damaged PC 12 cells through SIRT1/PGC-1alpha signaling pathway. |
