Acupoint-injecting can Raise Cortisol and Treat Asthma
 
   Junxiong Li* ,  Lao Changhui,  Yu yu,  Wu Qinghe,  Liang Keyun,  Zhang Hanmin, Tang Zhijun, Zheng Shengjie et al.
   Asthma Center, Guangzhou University of Traditional Chinese Medicine.  Teaching Hospital of Guangzhou University of Traditional Chinese Medicine.
                             Abstract
 Objective   Through both clinical and experimental study to inform that acupoint-injecting can raise body cortisol and  treat asthma.  Methods 
Ⅰ. clinic: 62 moderate asthmatics had been divided into 2 groups : 1) self-blood mixture globulin gamma acupoint-injecting group; 2) conventional treatment group.  At the beginning and the end of treatment, cortisol had been examined (cortisol had been tak
en at 8 am).  Ⅱ.  Experimentation : 51 guinea-pigs had been divided into 7 groups : 1) normal; 2) model; 3) conventional treatment ; 4) gamma globulin acupoint-injecting; 5) self-blood acupoint-injecting; 6) self-blood mixture gamma globulin acupoint-injec
ting; 7) self-blood mixture gamma globulin intramuscle. Like the study on human being , at the beginning and the end of the study , cortisol ,  cAMP/cGMP and beta receptor had been examined.  Each guinea-pigs group lung tissues had been examined under optic and electric microscope.   Results  notable change of cortisol had been seen at the end of study on both humans and guinea-pigs.  At the same time asthmatic symptoms  improved notably. It was improved that some inflammatory  cells had disappeared by examining under both microscopes examined .  Conclusion   acupoint-injecting can surely treat asthma probably owing raise cortisol and some reason.
Key words   Asthma, Self-blood, Gamma Globulin, Acupoint-injecting, Cortisol, Guinea-pig, Microscope.

 ( *Junxiong Li :302.28th Building of Nanhai Yard . Guangdong. P.R.China. 528200
     Fax: 86-757-6338098.  E-mail:doctorjli@21cn.com )
Introduction
  For a long time medical workers argue for ?if traditional medicine  really works in treating asthma?.  WHO/NHLBI?s GINA points out that ?In some countries traditional methods of healing are a primary way of treatment; in many countries, there has been a move toward using various traditional methods of healing. The scientific basis of these modes of therapy needs to be studied in detail, especially for countries in which these forms of therapy are frequently used.?.  In China, there are a great number of  doctors treating asthma by using traditional Chinese medicine while  using western or conventional medicine.  Actually they could decrease asthmatics symptoms and raise up their life quality.  Before less people improved  the  effective of  the methods of traditional Chinese medicine in treating asthma  by modern laboratory methods. Last year ?Acupuncture forum? held in NIH, USA.  Though there were still some difference opinion in acupuncture effective, a number of medical workers from many countries agreed with that acupuncture can treat a lot of disease including asthma.  And they agreed with that more research and study should be carrying out.  Nowadays more and more states in USA allow doctors and acupuncturists treat patients with acupuncture . A lot of studies on acupuncture are carrying out among many universities in different countries.
  We asthma center of Teaching Hospital of Guangzhou University of Traditional Chinese Medicine treated asthma by the method of acupuncture  for many years.  In order to improve the effective of the method , we had 4 years study on this programme. We were to compare the effective of acupuncture methods treating group and conventional  method treating group; comparing some laboratory test ; comparing changing under microscope.
 
Materials and Methods       SUBJECTS
The present study consisted of 2 part of studies: clinic and experimentation.  Clinical study was conducted in 62 asthmatics. Experimental study involved blood test like cortisol , Eos and pathological examination under optics and electric microscope.  62 patients with  a clinical history  of moderate asthma were recruited .  All the patients had been undergoing treatment for asthmatic  complaint in our asthma center for 1 year.  Regulatory permission and ethical committee approval was obtained .  The study was  carry out  under the Declaration of Helsinki  with all patients providing informed written or oral consent.
   For clinical study, a group patients treated by the method of acupoint-injecting with self-blood mixture globulin gamma consisting of 30 moderate asthmatics(treating group ; mean age 35.87) and another group treated with conventional method consisting of 32 asthmatics  (comparing group; mean age 39.14) were formed.  Both groups were matched with respect to the severity of the asthma.  The degree of severity of the asthma was determined on the basis of respiratory  complaints and anomalies in pulmonary function(FEV1).  For experimental Study,  51 guinea-pigs had divided into 7 groups : 1) normal; 2) model; 3) conventional treatment; 4) globulin treating; 5) self-blood acupoint-injecting; 6) self-blood mixture globulin gamma acupoint-injecting; 7) self-blood mixture globulin gamma intramuscle.   

Clinical study
STUDY DISIGN
The study was single ?blind and randomized in design, comparing 2 treatment methods. Two 1-week run-in periods were followed by a treatment period of 2 months and follow-up period of 2 weeks.
At the first visit  demographic details were recorded  and blood samples taken for determination of baseline haematological and biochemical parameters.
Treating group: Of these patients, male 15 and female 15.  mean age 35.87 years old. 
The injection :2ml vein self-blood  mixed with 1ml globulin gamma.
Acupoints: Extra 14, B11, B12, B13, S16 etc .
Course: Every 5 times as 1 course . Each time choose a pair of same name acupoints , each side inject 1.5 injection.  Every 2 days a time.  10 days later there were 2nd  course . And  ten days later there were 3rd course .  there were together 3 course treatments.
Examination: at the beginning and after  all courses  examined patients level of  Ig E, cortisol.
Conventional treating group
  Of these patients ,male14 and female 18, mean age 39.14 years old.
  Treating asthmatics as conventional treating methods( inhaled steroids, beta 2 agonists, theophyllin, antiallergic agent etc.
  Examination : also examine Ig E, cortisol and EOS at the beginning and after all the treating courses.
PROTOCOL
  Patients  issued  with daily record cards recorded asthma symptom scores and the use of additional rescue medication.  They also noted the frequency of nocturnal waking due to reversible airways obstruction.   At clinic visits demographic, clinical, biochemical and haematological details were recorded .  At each of these visits, independent assessments of the efficacy of treatment were made by the physician and patient were recorded in the physicians record form.
  At all visits the physician recorded any change in medication;, intercurrent illnesses, adverse events and patient withdrawals and heart rate and blood pressure were measured. 12-lead ECGs were performed at the beginning and after 3 months treatment.
STATISTICAL METHODS
  Analysis of percentage very effective and effective and changing of cortisol, Eos were analysed by Wilcoxon rank sum analysis.

    Table 1. Asthma symptom scores
Score                             Symptom
__________________________________________________________________                                                                                      
day-time
No symptoms during the day
Symptoms for one short period during the day
Symptoms for two or more short periods during the day
Symptoms for most of the day which did not affect normal daily activities
Symptoms for most of the day which did affect normal daily activities
Symptoms so severe that the patient could not go to work or perform normal daily activities
Night-time
No symptoms during the night
Symptoms causing waking once or early waking
Symptoms causing waking twice or more (including waking early)
Symptoms causing the patient to be awake for most of the night
Symptoms so severe that the patient did not sleep at all
________________________________________________________________

Results
      Acupoint-injecting with self-blood mixture globulin gamma produced a significantly greater improvement in the median day-time and night-time asthma score than conventional treatment.  The physicians and patients considered the treatment to be efficacious .
 Table 2. The comparing effective between the treating group and the conventional  treating group

 Inside
 the table the number is case(%) 
 Rate of very effective and effective: Treating group comparing conventional treating group  X=4.218  P<0.05

Table 3.  The changing of cortisol of 2 groups  （x±S）

─────────────────────────────────
groups              
   at the            after               P        
                       beginning        
   
                        （x±S）         （x±S） 
___________________________________________________________________
treating group         11.24±7.77       18.
23±11.80          <0.005  
conventional group     21.24±6.50       20.96±10.34          >0.05
─────────────────────────────────

Table 4.  The changing of EOS of 2 groups

─────────────────────────────────
group                 at the beginning       aft
er              P
                         （x±S）         （x±S）
─────────────────────────────────
treating               322.06±205.37    262.22±133.77       <0.005
conventional treating  331.54±163.35    281.88±107.21       <0.01

─────────────────────────────────

Table 5. The changing of Ig E of 2 groups(x±S)   IU/ml
────────────────────────────────
 group                  at the beginning          after             P
─────────────────────────────────
treating                421.80±210.15    375.10±260.
72       <0.05
conventional treating   445.86±261.25    405.16±198.53       >0.05        
─────────────────────────────────

ADVERSE EVENTS
   Besides little pain had been felt while injecting, no adverse events had been found.  Because of medical workers
 pay much attention to acupoint-injecting , no wounded of lung happened .

 
Experimental trial
  51 guinea-pigs, each weight 250 g, 25 male and 26 female.  All of them had randomized been divided into 7 groups.  
  1) Normal group:  injected  0.9% NS into abdomen of guinea-pigs.
  2) Model group:  made this group guinea-pigs became model according Hotson?s method.
Conventional treating group:  Each time acupoint-injecting with triamcinolonum  (0.225mg/guinea-pig).  
Globulin gamma group: acupoint-injecting with globulin gamma.
Self-blood group:  acupoint-injecting with self-blood .
Self-blood mixture globulin gamma acupoint-injecting group(treating group):  acupoint-injecting with self-blood mixture globulin gamma.
Self-blood mixture globulin gamma intramuscle group:  intramuscle with self-blood mixture globulin gamma
 
Table 6. The changing of the cortisol of the guinea-pigs
Group                           n             cortisol(
μg/dl)
                                              (x±s)     
  Normal                          8            33.92±15.43             
  Model                           8            15.50±7.73
  Conventional                      7            15.90±0.34

  Globulin gamma                   7            42.86±38.32
  Self-blood                        7            54.12±25.15
  Treating                          7            69.79±44.29
  Self-blood mixture globulin gamma im  7            59.44±22.87
__________________________________________________________________
Model vs Normal  P<0.01
Model vs Treating  P<0.001
Model vs Conventional P>0.05
Model vs Globulin gamma acupoint-injecting P>0.05
Model vs Self-blood acupoint-injecting P<0.01
Model vs Self-blood m
ixture globulin gamma im P<0.01

   Table 7. The  changing of  cAMP、cGMP and cAMP/cGMP of testing guinea-pigs（x±S）
─────────────────────────────────
Group                  n    cAMP（pm/ml） cGMP（pm/ml ） cAMP/cGMP
───────────────────────────────
（1）Normal  
         8  52.20±21.68*   13.04±2.07*     4.20±1.70**
（2）Model             8   33.67±8.80    15.50±2.63      2.18±0.51
（3）Conventional        7  62.05±7.85***   15.05±3.46    4.17±0.44***
（4）Globulin           7   33.23±12.84    14.93±2.75     2.16±0.52
（
5）Self-blood         7   44.96±16.88     14.62±7.69      3.20±1.35
（6）Treating           7   48.75±16.68*   14.75±5.04     3.59±1.58*
（7）S.B mixture G.G im  7   42.95±33.02      12.85±7.82     3.32±1.91
───────────────────────────────────
Compare to Model
group:
*    P<0.05
* *  P<0.01
***  P<0.001

  Table 8. Determination of beta receptor of guinea-pigs lung tissues of 3 groups (x±S）

Group                N   Bmax(fmol/mg pro.)         kd(nmol/L)

Model                 6     312.45±61.75          1.57±0
.76

Treating              9     401.87±113.33          2.03±0.89

Normal                7     452.40±103.23          2.22±0.92

Model group compare normal group，t=2.896, P<0.05；Model group compare treating group，t=1.752, P>0.05.                       
   

Observation under optics and electric microscope :
1. Optics microscope
  Got the samples from right bronchial and lung tissues of guinea-pigs.
Normal group: Bronchial membrane complete .  Pulmonary alveoli wall complete.  There were not exudate inside the cavity.  No Eos had been seen.
Model group:  (1) There were Eos and neutrophil mucus blots inside the cavity . Bronchial wall  hyperemia and oedema .  Smooth muscles of bronchioli were spasmed  and thicker.  (2) Pulmonary emphysema changing.  (3) There were consolidation inside pulmonary tissues .  There were lots of inflammation cells and a great number of RBC in pulmonary alveoli.            
Conventional group:  Small mucus blots could be seen inside the cavity . There were Eos and neutrophil on the bronchial wall. 
Globulin group: Compare model group, there were lass Eos.  There were scatter consolidation and focal pulmonary emphysema. 
Self-blood group: (1) It was not notable that Eos infiltration.  (2) Changing of pulmonary emphysema was focal emphysema. (3) Consolidation of lung were scatter consolidation.
Treating group:  (1) No mucus blots had been seen inside the cavity .  There were a little Eos on the bronchial wall.  (2) There were still scatter consolidation. 
Self-blood mixture globulin gamma intramuscle group: There were not  mucus blots inside bronchial wall.  Bronchial wall did not thicker distinctly .  There were a little  Eos on the bronchial wall.  Bronchioli lightly consolidation.     
Electric microscope
       Took 1 cubic mm of  tissues at tight lower part of guinea-pigs of each group.   Observed after treating by a special process. 
Normal group: Basement membrane between capillaries were not thicker.   Fibrous tissues were not hyperplasia.
Model group: There were various empty bubbles inside ciliated columnar epithelial cells.  Mitochondria were swollen.  Goblet cells were hyperplasia . Mucous glands were larger.  The explosive degranulation of most  of Eos were prominent.  Basement membrane were thicker.  Collagenous fibers increase.  Type II alveipar cells increased.
Conventional group: I type alveolar cells and Eos increased.  Fibrous tissues were hyperplasia.
Globulin group: There were still some Eos.  Fibrous tissues were not hyperplasia.
Self-blood acupoint-injecting group: There were still some Eos and II type alveolar cells and it was not distinct for increasing.
Treating group: There were not Eos .  Basement membrane were not thicker.   
Collagenous fibers were not increase . II type alveolar cells were basically normal
Self-blood mixture globulin gamma intramuscle group: There were not Eos .  Basement membrane were basically normal.  I and II type alveolar cells were basically normal.
Discussion
  Cortisol is endogenous hormone of human body.  It  changes regularly during 24 hours . It is highest at 8 am and lowest at night .  It is similar with regular changing of  ?YinYang ? theory of traditional Chinese medicine. Determination of cortisol can show the function of asthmatic hypothalamus-hypophysis-adrenal cortex and review serious stage of asthmatics.  A lot of asthma attacking happen during at deep night.  It is agree with degree of cortisol.  Degree of cortisol of many asthmatics especially serious patients or steroids-depend patients were lower than normal .  Those asthmatics used a great number of steroids so that their function of hypothalamus-hypophysis-adrenal cortex had been restrained . Their lives were easy to be harm.  Some acupuncture treatment studies reported that it can raise degree of cortisol.  It maybe the reason for acupuncture to treat asthma.
  At conventional treatment group , patients symptoms had been controlled but their degree of cortisol seldom raised.  If we can use endogenous hormone to decrease or instead of exogenic hormone we can surly raise up asthmatics? life quality and decrease their possibility of death.
  On experimental study, we also can find that some change happen on experimental guinea-pigs were agree with asthmatic patients.  The raising of cortisol of self-blood mixture globulin gamma group was highest.  The changing of lung and bronchial tissues of guinea-pigs examined under both optic and electric microscope show that inflammatory cells had disappeared  and it was similar with normal group.
  Both clinic and experimental study show that acupuncture can surly raise up degree of cortisol.  We think it is the one of the main reason that it can treat asthma.  We hope that we can work together with other medical research workers and repeat time and again our study in different ways .