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Effects of HapKok (LI-4) , SamUmGyo (SP-6) Acupuncture on Uterine Motility and Cyclooxygenase-2 Manifestation in Rats (합곡(合谷), 삼음교(三陰交) 자침(刺鍼)이 백서(白鼠) 자궁(子宮) 운동(運動) 및 Cyclooxygenase-2 발현(發現)에 미치는 영향(影響))

  • Lee, Byung-Chul;Lee, Ho-Sub;Kim, Kyung-Sik;Lee, Geon-Mok;Na, Chang-Soo;Kim, Jung-Sang;Hwang, Woo-Jun
    • Journal of Acupuncture Research
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    • v.17 no.2
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    • pp.187-208
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    • 2000
  • By the activation of ovary hormone, many morphological changes occur in the epithelial cell lines and muscle cells in rat uterus. These two cells in uterus are important to the implantation of embryo, maintaining pregnancy and starting parturition. One important change associated with the morphological change of these two cells in uterus is the change on prostaglandin(PG) metabolism. Its presence and synthesis in endometriurn and myometrium in uterus affects estrous cycle and the start of embryo implantation in uterus. It also performs as an important modulator in parturition. So the abnormally weak expression of PG causes difficulty during labor and over-expression causes pre-term labor. PG biosynthesis starts from either free or liberated arachidonic acids from membrane phospholipid by phospholipase. Such arachidonic acids are converted into PG catalyzed by Cyclooxygenase. Under normal physiological condition, Cyclooxygenase-1(COX-1) having 602 units of amino acids controls the synthesis of PG. It acts as a local hormone regulating vasomodulation of blood flow, flexible muscle movement, increasing the blood permeability and contributing the protective role in preserving integrity of the stomach lining and Cyclooxygenase-2 (COX-2) is induced by the inflammation, pregnancy and increased its expression until parturition. Lipid metabolite like PG is located in uterine and expression of COX-2 increased with pregnancy. Increased expression of COX proteins in epithelial cells and myometrial cells are told to increase the muscle contractility in uterus but decreased right after the labor in rat. It is a good sign indicating that COX proteins are deeply related to the start of labor. Currently, Several studies report the use of PG and COX-2 inhibitor as medication for controlled abortion or to prevent pre-term labor but they entail various side-effects. Our study proposed to suggest use of acupuncture as an another mediator to control abortion or pre-term labor without causing unnecessary side-effects by those medicines. Two acupuncture sites, LI-4 & SP-6 were selected due to their known efficacy. From the immunohistochemical staining of COX-2, normal expression of COX-2 protein in nonpregnant SD rat's uterus revealed that COX-2 protein was primarily detected in the lumina epithelial lining and in the epithelial cell lining contacting the stromal cells. High resolution optical microscopic scanning revealed distinguishable staining in the myometrial mucosa. LI-4 acupuncture administered nonpregnant rat's uterus showed strong expression for COX-2 in endometrium contacted with lumina epithelial lining of rat uterus and in myometrial mucosa. Stromal cells showed more staining than untreated nonpregnant rat's uterus and stronger staining in stromal cells contacting myometrial layer compared to untreated nonpregnant rat's uterus. SP-6 acupuncture administered nonpregnant rat's uterus showed weak expression for COX-2 in myometrial layers and stromal cells but no staining was visible in lumina epitheliai and glandular epithelial cells. Few stromal cells and myometrial mucosa were positively stained for COX-2. Pregnant SD rat's uterus was also immunostained for COX-2 expression after 18 days of pregnancy. Unlike to untreated nonpregnant rat's uterus, luminal epithelial cells were not positively stained for COX-2 but stronger staining for COX-2 was revealed in stromal cells. LI-4 acupunctured SD rat's uterus had very strong expression of COX-2 in luminal epithelial lining. Few stromal cells showed stronger positive COX-2 staining and myometrial layers also showed more expression than untreated pregnant rat. SP-6 acupuncture administered pregnant SD rat's uterus showed positive expression of COX-2 in epithelial cells of luminal mucosa layer but weaker than that of LI-4 acupuncture treatment's case. However, strong positive staining was revealed in stromal mucosa and myometrial layers. Virgin SD rat's uterus motility index during LI-4 acupuncture was 66.52 % (Prob〉T = 0.0197) compared to its motility before the acupuncture treatment but the motility index was slighdy elevated up to 79.58 % (Prob〉T = 0.1175) after the acupuncture. During the SP-6 acupuncture treatment for 30 minutes, uterus motility index was 90.52 % (Prob〉T = 0.1832) showing lesser decrement but consequently reached similar motility index decreasal to 79.95 % (Prob〉T = 0.0215) after the acupuncture treatment as LI-4 showed. LI-4 acupuncture tend to be a quick treatment to reducing the uterus motility in a virgin rat but eventually both two acupuncture administration created very similar reduction of uterus motility seeing the index after the both acupunctures. The uterus movement monitored during the LI-4 acupuncture administered for 30 minutes, Pregnant SD rat showed decreased motility down to 77.90 % (Prob〉 T = 0.0076) compared to uterus motility before the acupuncture and it continuously decreased down to 71.81 %(Prob〉T = 0.0214) after the removal of needle. The statistical analysis using paired t-test showed significance difference for both two motility indexs at =0.05. SP-6 acupuncture administered to pregnant SD rat also had similar pattern of decreasing uterus motility index down to 74.70 % (Prob〉T = 0.1730) during the initial 30 minutes acupuncture administration and it was continuously lowered to 71.52 % (Prob〉T = 0.0155) after the acupuncture. The paired t-test resuit for SP-6 suggest prompt response of uterus motility index to the SP-6 acupuncture treatment but consequently reached same level of inducing the motility reduction as LI-4 at =0.05 level.

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STUDIES ON THE DIMORPHISM AND FERTILITY OF PERSICARIA JAPONICA (MEISSNER) GROSS ET NAKAI (Persicaria Japonica (MISSNER) Gross et Nakai의 이형화와 수정력에 관한 연구)

  • HARN, Chang Yawl
    • Journal of Plant Biology
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    • v.3 no.1
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    • pp.1-15
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    • 1960
  • HARN, Chang Yawl : Studies on the dimorphism and Fertility of Persicaria japonica (MEISSNER) Gross et Nakai. Kor Jour. Bot. 3(I) 1-15 1960 Numerous investigations, since the works of DARWIN, have been made regarding the heterostylous plants by JOST (1907), CORRENS (1924), LAIBACK (1924), LEWIS (1943), and many others. Studies on the heterostylous Polygomum, however, were not reported except for the buckwhent, Fagopyrum esculentum, which was investigated by SCHOCH-BODMER (1930), EAST (1934), FROLOVA & Co-Workers (1946), MORRIS (1947, 1951) TATEBE (1949, 1951, 1953), present author (1957), and others. It is because no heterostylous species, besides buckwheat, have been known to exist in the Polygonum family. The author, during his studies on both heterostylism and fertility of Polygonaceae, has found that the species, persicaria japonica (Meissner) Gross et Nakai, is not diecious as has been known in taxonomy, but in reality beterostylous both morphologically and physiologically. It was found that this plant, regarded by taxonomist, as a male plant setting no seed, actually set seed (botanical fruit) when legitimate combination was made. Since his brief report on the dimorphic phenomens of this plant in 1956, the author's further research on the manner of fertilization has revealed that this species is a peculiar type whose dimorphism has undergone extreme specialization structurally and physiologically, the short-styled individual behaving in nature as a male plant and the long-styled individual, as female, whereas in controllled pollination the plant shows highly differentiated typical dimorphism. When compared with the other dimorphous species of this family, F. esculentum and P. sentiosa. it has been clarified that these three species differ in the degree of differentiation of their dimorphism morphologically and physiologically. That is, P. japonica has developed such a high specialization as to mislead the taxonomists, while P. senticosa shows almost no noticeable difference between long- and shortstyled individuals retaining most of the inherent physiological character cmmon to the genus except for the fact that it has two forms of flowers. F. esculentum appears to have taken the intermediate position in every respect. The result obtained in the present experiment are summarized as follows: 1) P. japonica has two kinds of individuals, one long style-short stamened; the other, short style-long stamened. The floral structure of this plants shows typical characteristics of dimorphic heterostylism. The differentiation between the two forms of flower has proceeded so highly both in primary and secondary difference of flower structure that this may be regarded as the most specialized form of dimorphism. 2) The differences of floral structure between the long and short styled individuals are remarkable compared with the other dimorphic species of the family. 3) The stamens of long styled plants show the sign of deteriolation whereas those of the short styled flower are well-developed. 4) When legitimate combinations are made, both L- and S-styled individuals are fertilized well and set seed (fruit), while in the illegitimate combination no fertilization and seed setting occur. Physiologically this species exhibits the typical behavior of dimorphic plants. 5) The self-fertile character, so common in other species of the other non-heterostyle Polygonum family, has disappeared completely. 6) Under natural conditions, no or few seed setting is observed in short styled individuals that behave as if they were male plants. 7) In hand pollination, the combination of both $L{\times}S$ and $S{\times}L$ alike yield relatively good fertility and seed-formation, the behavior of short styled individuals in artificial pollination differing remarkably from that in nature. 8) Under controlled pollination, $L{\times}S$ combination sets far more seed than in the combination of $S{\times}L$. In the S-styled individuals, the fertilized flower has the tendency of its seed more readily falling off in every stage of seed development than in the L-styled individuals. 9) The behaviors of pollen tubes just parallels the results of fertility test. That is, in the illegitimate combination, L-selfed, $L{\times}L$, S-selfed, and $S{\times}S$, the growth of pollen tubes is checked in the style, while in legitimately combined $L{\times}S$ and $S{\times}L$, the pollen tubes grow well reaching the ovaries within 40-50 minutes after pollination. The response of short styled individuals, known as male plant among taxonomists, is identical, as far as behavior fo pollen tube growth and fertilization are concerned, to that of long styled individuals, the so-called female plant. 10) The pollen grains from the short-styled plants are complete and fertile, whereas 70% of those of L-styled are found to be abortive, i.e., empty contents. 11) The remaining 30% of pollen of L-plant shows varied degree of stainability when stained with iron-aceto-carmine......mostly light red, while the pollen grains of S-style individuals are dark brown indicating complete fertility and viability. 12) The abundance of sterile pollen in L-styled and the nature of seed-dropping which occurs in S-styled individuals appear to be the main causes why the short styled individuals bear no seed in nature. Under controlled legitimate union, $S{\times}L$, the careful and elaborate pollination would give the S-styoled flowers the opportunities to receive the fertile pollens, though few in number, from L-styled plant, thus enabling S-plant to bear seed. 13) This species is not dioecious as is regarded by taxonomists, but typical dimorphic plant which has so highly specialized in floral structures and funcitons that the long-styled plant behaves just like a female individual; and the short-styled, like a male.

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